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2018

Printing of platinum for use in neural implants

Ryan Patrick Sullivan
University of Wollongong

Follow this and additional works at: https://ro.uow.edu.au/theses1

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Recommended Citation
Sullivan, Ryan Patrick, Printing of platinum for use in neural implants, Doctor of Philosophy thesis,
Australian Institute for Innovative Materials, University of Wollongong, 2018. https://ro.uow.edu.au/
theses1/261

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 PRINTING OF PLATINUM FOR USE IN NEURAL
IMPLANTS

A thesis submitted in fulfillment of the requirement for the award of the degree

DOCTOR OF PHILOSOPHY
from
THE UNIVERSITY OF WOLLONGONG

by

RYAN PATRICK SULLIVAN

Supervisors:

Prof. Gordon G. Wallace

A/Prof. Michael J. Higgins

Dr. Zhillian Yue

Australian Institute of Innovative Materials

June 2018
 CERTIFICATION

I, Ryan Patrick Sullivan, declare that this thesis, submitted in fulfillment of the requirements
for the award of Doctor of Philosophy in the Australian Institute of Innovative Materials at
the University of Wollongong, is entirely my own work unless otherwise referenced or
acknowledged. This document has not been submitted for qualifications at any other
academic institution.

Ryan Patrick Sullivan

20 June 2018

Ryan P. Sullivan i
 PUBLICATIONS

The following publications and presentations have arisen directly from work contained

within this thesis.

Publications in Refereed Journals:

 O’Connell C.D., Higgins M. J., Sullivan R. P., Moulton S.E., Wallace, G.G. Ink-on-

Probe Hydrodynamics in Atomic Force Microscope Deposition of Liquid Inks. Small.

2014; 10(18) 3717–3728; doi: 10.1002/smll.201400390.

Author contribution: I assisted with the mathematical analysis and interpretation of

data, along with development of the three regime model.

 O’Connell C.D., Higgins M. J., Sullivan R.P., Jamali S.S., Moulton S.E., Wallace, G.G.

Nanoscale platinum printing on insulating substrates. Nanotechnology; 2013;

24(50):505301; doi:10.1088/0957-4484/24/50/505301

Author contribution: I assisted with the design of plasma reduction parameters and

performed all SEM and SEM-EDS measurements of deposited features.

 Wagner M, O’Connell C.D., Harman D.G., Sullivan R., Ivaska A., Higgins M.J.,

Wallace G.G., Synthesis and optimization of PEDOT:PSS based ink for printing

nanoarrays using Dip-Pen Nanolithography. Synthetic Materials; 2013; 181 64-71.

Author contribution: I fabricated the PEDOT:PSS and component patterns and

analyzed FTIR data.

Ryan P. Sullivan ii
Manuscripts in Preparation:

 Sullivan, R.P., Higgins, M.J., Yue, Z., Wallace, G.G. “Rapid Automatic

Characterization of Inkjet Grids Using an ImageJ Plugin.”

 Sullivan, R.P., Higgins, M.J., Yue, Z., Wallace, G.G. “Inkjet Printing of Conductive

Platinum Lines on PDMS for use in Neural Implants.”

 Sullivan, R.P., Higgins, M.J., Yue, Z., Wallace, G.G. “Rapid Inkjet Deposition of

Platinum using 10 pL Polyol Ink Drops.”

 Sullivan, R.P., Higgins, M.J., Yue, Z., Wallace, G.G. “Dip-Pen Nanolithography of

Platinum on PDMS: Difficulties of Soft, Hydrophobic Substrates.”

Conference Proceedings:

 Sullivan, R.P., Higgins, M.J., Yue, Z., Wallace, G.G. Micronscale Inkjet Printing and

Nanoscale Dip-Pen Nanolithography of Platinum on PDMS towards use in Bionics

fabrication. The 10th Annual International Electromaterials Symposium.

Wollongong NSW, Australia February 11-13, 2015.

 Sullivan, R.P., Higgins, M.J., Yue, Z., Wallace, G.G. Platinum Printing on PDMS. The

9th Annual International Electromaterials Symposium. Wollongong NSW, Australia

February 12-14, 2014.

Ryan P. Sullivan iii

  Sullivan, R.P., Higgins, M.J., Yue, Z., Wallace, G.G. Platinum Printing on Flexible

Insulating Substrates. The 4th Asia-Pacific Symposium on Nanobionics, Melbourne

VIC, Australia November 14-15, 2013.

 Sullivan, R.P., Wagner, M., O’Connell, C.D., Nakashima, H., Higgins M.J., Wallace,

G.G. Dip-Pen Nanolithography of PEDOT:PSS: Capabilities and Prospects. The 8th

Annual International Electromaterials Science Symposium, Wollongong NSW,

Australia February 13-15, 2013.

 O’Connell, C.D., Sullivan, R., Higgins, M.J., Moulton, S.E., Wallace, G.G. Nano-scale

electroless platinum printing: micro-snowflakes and nano-dots. The 8th Annual

International Electromaterials Science Symposium, Wollongong NSW, Australia

February 13-15, 2013.

 Sullivan, R.P., O’Connell, C.D., Yue, Z., Higgins, M.J., Wallace, G.G. Using Dip-Pen

Nanolithography for fabrication of the next generation of flexible bionic electrodes:

that of the nanoscale regime. The 3rd Asia-Pacific Symposium on Nanobionics,

Wollongong NSW, Australia September 19-21, 2012.

 O’Connell, C.D., Sullivan, R., Sherrell, P., Jamali, S., Higgins, M. J., Moulton, S.E.,

Wallace, G.G. Electroless Nanoscale Noble Metal Printing on Flexible Substrates

with Dip Pen Nanolithography. The 3rd Asia-Pacific Symposium on Nanobionics,

Wollongong NSW, Australia September 19-21, 2012.

Ryan P. Sullivan iv
 Yue Z., Molino M.P., Liu X., Sullivan R., Svehla M., Wallace G.G. Surface

Bioengineering Strategy to Improve Electrode Performance. The 2nd International

Conference on Medical Bionics, Melbourne VIC, Australia November21, 2011.

Additional Publications

The following publications and presentations have arisen as a result of other projects that I

have been involved in during my doctoral studies, but are unrelated to this thesis.

Publications in Refereed Journals:

 Andrews J.L, Sullivan R.P., Wang H., Newell K.A., Huang XF., Fernandez-Enright F.

Pharmacological blockade of Lingo-1 in combination with olanzapine

administration reverses phencyclidine induced effects on dendritic morphology,

cognitive performance and locomotor activity. (In Preparation).

Author contributions: I designed the confocal microscopy protocol and created

ImageJ scripts and protocol to help analyze the dendrites and spines. I fed and

injected rodent specimens on weekends.

 Andrews J.L., Zalesky A., Nair S.S., Sullivan R.P., Colson, N.J., Clarke S.J., Green M.J.,

Pantelis C., Newell K.A., Fernandez-Enright F. Genetic and epigenetic regulation in

Lingo-1: effects on cognitive function in a case-control study for schizophrenia.

(Submitted to Molecular Psychiatry)

Author contributions: I assisted with statistical design and implementation of

statistical analyses and interpretation of the results.

Ryan P. Sullivan v
Conference Proceedings:

 Andrews J.L., Sullivan R.P., Newell K.A., Huang XF, Fernandez-Enright F.

Pharmacological blockade of Lingo-1 in combination with olanzapine

administration reverses phencyclidine induced effects on dendritic morphology,

cognitive performance and locomotor activity. The 45th Annual Meeting of the

Society for Neuroscience, Chicago IL, USA October 17-21, 2015.

 Andrews J.L., Sullivan R.P., Newell K.A., Huang XF, Fernandez-Enright F. Effects of

pharmacological blockade of Lingo-1 pathways in a Phencyclidine Rat Model for

Schizophrenia. The 44th Annual Meeting of the Society for Neuroscience,

Washington DC, USA November 15-19, 2014.

Ryan P. Sullivan vi
 TABLE OF CONTENTS

CERTIFICATION ..................................................................................................... I
PUBLICATIONS ..................................................................................................... II
TABLE OF CONTENTS.......................................................................................... VII
LIST OF FIGURES ................................................................................................. XI
LIST OF TABLES .................................................................................................. XX
LIST OF ABBREVIATIONS .................................................................................. XXII
ABSTRACT ...................................................................................................... XXIII
ACKNOWLEDGEMENTS ................................................................................... XXVI
CHAPTER ONE: INTRODUCTION AND REVIEW OF THE LITERATURE ....................... 1
1.1 INTRODUCTION TO NEURAL PROSTHETICS .................................................................. 1
1.1.1 History of Prosthetics ....................................................................................... 1
1.1.2 Neural Interfacing Techniques ......................................................................... 3
1.2 TYPES OF NEURAL ELECTRODES AND LOCATIONS SITES .................................................. 8
1.2.1 Hearing ............................................................................................................. 8
1.2.2 Vision .............................................................................................................. 12
1.2.3 Arms and Legs ................................................................................................ 17
1.2.4 Memory.......................................................................................................... 19
1.3 ELECTRODE MATERIALS AND DESIGN ...................................................................... 20
1.3.1 Overview of Implant Challenges .................................................................... 20
1.3.2 General Electrode Design............................................................................... 23
1.3.3 Insulating materials ........................................................................................ 25
1.3.4 Impact Resistance/Structural Layer ............................................................... 26
1.3.5 Adhesion Layer ............................................................................................... 27
1.3.6 Conductive Layer and Electroactive Surface .................................................. 28
1.3.7 Biologics ......................................................................................................... 31
1.4 FABRICATION TECHNIQUES................................................................................... 32
1.4.1 Deposition Overview ...................................................................................... 36
1.4.2 Dip-Pen Nanolithography............................................................................... 39
1.4.3 Inkjet .............................................................................................................. 42
1.4.4 Metal Inks and the Polyol Process ................................................................. 45
1.5 RATIONALE ...................................................................................................... 56
1.6 AIMS AND OBJECTIVES ........................................................................................ 56
1.6.1 General Aim ................................................................................................... 56
1.6.2 Specific Aims .................................................................................................. 57
1.7 SIGNIFICANCE ................................................................................................... 57

CHAPTER TWO: DIP-PEN NANOLITHOGRAPHY OF PLATINUM INK ONTO

POLYDIMETHYLSILOXANE .................................................................................. 59

Ryan P. Sullivan vii

 2.1 INTRODUCTION ................................................................................................. 59
2.2 EXPERIMENTAL DETAILS ...................................................................................... 60
2.2.1 PDMS Substrate Creation............................................................................... 60
2.2.2 Polydopamine Synthesis ................................................................................ 64
2.2.3 Ink Formulation .............................................................................................. 64
2.2.4 DPN Modification ........................................................................................... 65
2.2.5 DPN Fabrication ............................................................................................. 67
2.2.6 DPN Feature Characterization ....................................................................... 68
2.3 RESULTS AND DISCUSSION ................................................................................... 69
2.3.1 PDMS Casting ................................................................................................. 69
2.3.2 PDMS Surface Modification with Dopamine ................................................. 71
2.3.3 DPN Deposition .............................................................................................. 73
2.3.3.1 NanoInscriptor to Asylum ...................................................................................... 73
2.3.3.2 Effect of Tip Modification ...................................................................................... 73
2.3.3.3 Effect of Withdrawal Speed ................................................................................... 75
2.3.3.4 Line Printing on PDMS-PDA without Plasma Reduction ........................................ 78
2.3.3.5 Line Printing on PDMS-PDA with Plasma Reduction ............................................. 82
2.3.4 Scanning Electrochemical Microscopy ........................................................... 83
2.4 CONCLUSION .................................................................................................... 84
2.5 APPENDIX ........................................................................................................ 85
2.5.1 Overlapping drops.......................................................................................... 85

CHAPTER THREE: AUTOMATION OF CHARACTERIZATION OF INKJET DROPLETS

USING IMAGEJ SOFTWARE ................................................................................. 87
3.1 INTRODUCTION ................................................................................................. 87
3.2 EXPERIMENTAL DETAILS ...................................................................................... 90
3.2.1 Geometric and Order Values ......................................................................... 91
3.2.2 Overview of program ..................................................................................... 94
3.2.2.1 Calibration Grids .................................................................................................... 94
3.2.2.2 Installation and Usage ........................................................................................... 94
3.3 RESULTS AND DISCUSSION ................................................................................... 99
3.3.1 Automatic Background Removal.................................................................... 99
3.3.2 Binarization .................................................................................................. 102
3.3.3 Sorting .......................................................................................................... 103
3.3.4 Example Data Set ......................................................................................... 108
3.4 CONCLUSION .................................................................................................. 110
3.5 APPENDIX ...................................................................................................... 111
3.5.1 Source Code for Inkjet Plugin ....................................................................... 111

CHAPTER FOUR: 10 PICOLITRE INKJETTING OF PLATINUM INKS ........................ 129

4.1 INTRODUCTION ............................................................................................... 129
4.2 EXPERIMENTAL DETAILS .................................................................................... 131
4.2.1 Ink Formulation ............................................................................................ 131
4.2.2 In Flight Jetting ............................................................................................. 132
4.2.3 PDMS Substrate Creation............................................................................. 133

Ryan P. Sullivan viii

 4.2.4 Ink Jetting of Grids ....................................................................................... 135
4.2.5 Reduction of H2PtCl6 .................................................................................... 135
4.2.6 Conductive AFM ........................................................................................... 136
4.2.7 SEM-EDS ....................................................................................................... 137
4.3 RESULTS AND DISCUSSION ................................................................................. 137
4.3.1 In Flight Jetting Analysis ............................................................................... 137
4.3.1.1 Drop Volume........................................................................................................ 137
4.3.1.2 Drop Velocity ....................................................................................................... 141
4.3.1.3 Drop Coalescence ................................................................................................ 142
4.3.2 Inkjetted pattern analysis ............................................................................ 143
4.3.3 Morphology of Features based on Reduction ............................................. 149
4.3.4 Penetration Depth of Dropcast Films........................................................... 154
4.3.5 Conductivity ................................................................................................. 156
4.3.6 Elemental Composition ................................................................................ 160
4.3.7 Ink-Inkjet Head Interactions......................................................................... 163
4.4 CONCLUSIONS ................................................................................................. 164
4.5 APPENDIX ...................................................................................................... 169
4.5.1 Vapor Pressure of Ink ................................................................................... 169
4.5.2 Error ............................................................................................................. 170
4.5.3 Conductivity ................................................................................................. 170

CHAPTER FIVE: 1 PICOLITRE INKJETTING OF PLATINUM INKS ............................ 172

5.1 INTRODUCTION ............................................................................................... 172
5.2 EXPERIMENTAL DETAILS .................................................................................... 174
5.2.1 Ink Creation, In-Flight Jetting, and PDMS Substrate Creation ..................... 174
5.2.2 Inkjetting of Grids......................................................................................... 174
5.2.3 PDMS Plasma Modification .......................................................................... 175
5.2.4 PDMS Modification with Polydopamine ...................................................... 175
5.2.5 Inkjetting of Lines ......................................................................................... 176
5.2.6 Multimeter Conductivity .............................................................................. 176
5.2.7 Scanning Electrochemical Microscopy ......................................................... 176
5.2.8 Conductive AFM ........................................................................................... 176
5.2.9 Sonication Adhesion Test ............................................................................. 177
5.3 RESULTS AND DISCUSSION ................................................................................. 177
5.3.1 In Flight Jetting Analysis ............................................................................... 177
5.3.1.1 Drop Volume........................................................................................................ 177
5.3.1.2 Drop Velocity ....................................................................................................... 179
5.3.1.3 Drop Coalescence ................................................................................................ 179
5.3.2 Inkjetted pattern analysis ............................................................................ 180
5.3.3 Substrate Modification ................................................................................ 184
5.3.4 N2 Plasma Reduction .................................................................................... 189
5.3.5 Conductivity ................................................................................................. 189
5.3.6 Polydopamine film ....................................................................................... 190
5.4 CONCLUSION .................................................................................................. 192
5.5 APPENDIX ...................................................................................................... 194

Ryan P. Sullivan ix
 5.5.1 Calculating the effect of pixel resolution on circularity and roundness ...... 194
5.5.2 Cochlear CI512 Electrode Calculations ........................................................ 196
5.5.3 Source code for Analyze Stripes Plugin ........................................................ 197

CHAPTER SIX: CONCLUSIONS AND OUTLOOK ................................................... 205

6.1 OVERALL FINDINGS .......................................................................................... 205
6.2 UPSCALING AND DOWNSCALING POLYOL INKS ........................................................ 207

REFERENCES .................................................................................................... 211

Ryan P. Sullivan x
 LIST OF FIGURES

Figure 1-1: Ancient Prosthetic devices. An ancient false toe from an Egyptian
mummy (Finch, 2011), and drawings of the iron hands of Gottfried von Berlichingen
(Mechel, 1815). Reprinted from Finch, J., (2011) The ancient origins of prosthetic
medicine; The Lancet; 377(9765), 548-549, with permission from Elsevier. Copyright
© 2011 Published by Elsevier Ltd. ............................................................................... 2

Figure 1-2: The first cochlear implant. A photograph of the UMDOLEE multiple
electrode prosthesis for implantation next to a 20 cent piece. Reprinted from Clark
GM, Pyman BC, Bailey QR., The surgery for multiple-electrode cochlear
implantations. J Laryngol Otol. 1979 Mar;93(3):215-23 with permission from
Cambridge University Press. ........................................................................................ 3

Figure 1-3: Prosthetic hand in eight finite-states. Reprinted with permission from
Dalley, S.A., Bennet, D.A., Alshammary N.A., Goldfarb M. Multigrasp Hand
Prosthesis and Myoelectric Control Method for Enhancing the Functional Capability
of Upper Extremity Amputees; 2012; IEEE Life Sciences; Copyright © 2016, IEEE. .... 5

Figure 1-4: Three types of recording devices for neural interfacing;

electroencephalography (EEG), electrocorticography (ECoG), and microelectrode
array (MEA). EEG consists of many electrodes placed along the scalp or neck. ECoG
consists of an electrode array placed on the surface of the brain. MEAs penetrate
the outer brain tissue, allowing them to have more localized recording than either
EEG or ECoG. ................................................................................................................ 6

Figure 1-5: Schematic representation of sound transduction and anatomy of the

cochlea. Sound waves focused from the outer ear travel through the ear canal to
the tympanic membrane. The perturbations are then transferred through the three
ossicles, the malleus, incus, and stapes, to the oval window. ..................................... 9

Figure 1-6: Schematic representation of nerve impulse transduction pathway to the

auditory cortex. Hair cells in the cochlea stimulate the cochlear nerve, which
projects to the cochlear nuclei. From here, signals propagate to the Inferior
Colliculus, the medial geniculate nucleus, and finally the primary auditory cortex. 10

Figure 1-7: Schematic representation of a cross–section of an eye. Light travels

through the cornea, pupil, lens and projects onto the back of the retina. The retina
(bottom panel) contains photoreceptors for color vision (cones) and low light, or

Ryan P. Sullivan xi
black and white vision (rods). There are 3 sites for visual implants within the retinal
layers: epi-retina, sub-retina, and supra-choroid (top panel). .................................. 12

Figure 1-8: Schematic representation of the neural pathways for vision. From the
retina, neural signals for vision travel along the optic nerve, the optic tract, synapse
on the lateral geniculate nucleus in the thalamus and then project to the visual
cortex. The different sites for the 5 types of visual implants or stimuli are marked. 14

Figure 1-9: A. Corticospinal tracts – descending pathway originating in the motor

cortex, responsible for motor control of the proximal and distal arms and legs. B.
Dorsal columns-medial lemniscus pathway – ascending pathway originating in the
limbs, responsible for transmitting discriminative touch and proprioception
feedback from the proximal and distal arms and legs. (Reprinted from
TeachMeAnatomy.com (http://teachmeanatomy.info/neuro/pathways/ascending-
tracts-sensory/ and http://teachmeanatomy.info/neuro/pathways/descending-
tracts-motor/) © 2015-2016 TeachMeAnatomy.com, under the Creative Commons
Attribution-NonCommercial-No Derivatives 4.0 International license
(https://creativecommons.org/licenses/by-nc-nd/4.0/). .......................................... 18

Figure 1-10: Schematic of Hippocampal implants interfacing between CA3 and CA1.
Due to lateral symmetry, there are two mirrored hippocampi, and therefore two
mirrored implants. ..................................................................................................... 20

Figure 1-11: Schematic cross-section of theoretical neural implant. It minimally

consists of an insulating substrate and conducting layer, which may double as the
electroactive surface. Impact layers may be added to facilitate insertion if the
substrate is overly soft. An adhesion layer is used if there are issues with adhesion
between the conducting layer and the insulating substrate. Coatings for
electroactive surfaces have been of interest for their ability to drastically increase
the charge injection capacity. .................................................................................... 24

Figure 1-12: Logarithmic Scale Graph of Elastic Modulus of various implant

materials and target tissues. Reprinted from Scholten and Meng; Materials for
microfabricated implantable devices: a review; 2015; Lab on a Chip; with
permission from the Royal Society of Chemistry. Copyright © 2015, Royal Society of
Chemistry. .................................................................................................................. 25

Figure 1-13: Chemical structures of PDMS, Parylene C and Polyimide. .................... 26

Figure 1-14: (L-R): Smooth platinum, Platinum Gray, and Platinum Black. Scale: 10
μm. Reprinted with permission from: Zhou, D.D., Dorn, J.D., Greenberg, R.J.; The

Ryan P. Sullivan xii

Argus® II retinal prosthesis system: An overview; 2013 IEEE International
Conference on Multimedia and Expo Workshops, © 2013 IEEE. .............................. 31

Figure 1-15: Scale of DPN and Inkjet fabrication capabilities based on volume of
droplets deposited. The number in parenthesis is the diameter of a drop at the
given volume to give an estimate of the resolution. The diameter will naturally
change with contact angle based on the ink and substrate used. ............................ 39

Figure 1-16: DPN ink transfer. 1) Tip between features; 2) contact with substrates
causes meniscus to form. Diameter of features is proportional to the square root of
dwell time. 3) Rupture of capillary bridge determines the volume of a sphere during
retraction (O’Connell et al., 2014); 4) Deposited feature takes upon the form of a
spherical cap after reaching equilibrium. .................................................................. 40

Figure 1-17: Schematic of the interior of single piezoelectric inkjet chamber. ........ 43

Figure 1-18: Schematic representation of the three types of metal inks. Since this
thesis work has focused on platinum, the schematic representation is of the 3
different types of platinum inks that are possible. .................................................... 46

Figure 1-19: Mechanism of plasma reduction and sintering of salt inks................... 49

Figure 1-20: Table of elements with metals labeled by viability of use in polyol
process. Elements in green have been demonstrated by create reduced films.
Elements in yellow have been shown to reduce to nanoparticles, though no work
has been done to create films. Elements in blue reduce to metal oxide are therefore
not viable without further steps. ............................................................................... 51

Figure 2-1: Graphical representation of the steps required to prepare PDMS

samples for subsequent fabrication. ......................................................................... 63

Figure 2-2: Aging inks were found to be a problem if kept over periods of months.
The left picture is of bulk ink creation. The right picture has two inks; the left ink
was made 10min before the image was taken while the right ink was made 6
months before the image was taken. ........................................................................ 65

Figure 2-3: Picture of 12 DPN tips on a cantilever. The distance between tips, 66
μm, was an important factor in printing. No pattern was created wider than 50 μm
so as to remove possible interactions between the adjacent tip and already
deposited features. .................................................................................................... 66

Ryan P. Sullivan xiii

Figure 2-4: Optical images of a DPN array using the internal camera of an Aslyum
AFM. The left image shows the cantilever leveled so that no tips touch while the
right image shows the cantilever leveled so that when the tip closest to the bottom
of the screen touches the substrate, the other tips are less deflected. .................... 67

Figure 2-5: Pictures of two petri dishes filled with PDMS precursors. (A) PDMS
precursor solution is cloudy white, due to extra surfactants on the glass surface
going into solution. (B) PDMS precursor solution is clear as required. ..................... 70
PDMS samples were cured at temperatures of 80°C, 100°C, and 120°C for 3 hours. It
has been shown that the silicon hydride in PDMS precursor curing agents is able to
reduce metal salts such as AgNO3, HAuCl4, and H2PtCl6 ............................................ 70

Figure 2-6: AFM image of a partially cured PDMS substrate that has adhered to the
DPN tip, causing large deformations as the PDMS was raised when withdrawing the
tips from the substrate .............................................................................................. 71

Figure 2-7: Optical images of (A) DPN tip array and the edge of the chip; and (B)
zoomed in image of the left most chip in panel A. These images show that the metal
ink does not run off the tip unlike previously seen in our past work (O’Connell et al.,
2013). We attribute this to the fact that we had previously used plasma cleaning in
order to reuse the tips. In this work, all tips were single use only. Both unmodified
and ODTMS modified DPN tips showed that ink remained on the tips with no
evidence for any on-chip migration over all of the arrays surveyed. ........................ 74

Figure 2-8: Effect of withdrawal speed on deposition with DPN. Images A, B, and C
are profiles of single features in D, E, and F respectively. 1 μm/s approach and
withdrawal speed produces no deposition as can be seen in panels A and D. 10
μm/s approach and withdrawal speed produces deposition as seen in panels B and
E. 100 μm/s approach and withdrawal speed causes indentation and deformation of
the PDMS surface as seen in panels C and F. Scale bars 10 μm. ............................... 77

Figure 2-9: AFM images taken in AC mode of grids printed on PDMS-PDA substrate.
A-C are height profiles of D-F. Panels A and D are the profile and height map of a
representative grid printed with 2s dwell 10 nN applied force. Panels B and E are
the profile and height map of a representative grid printed with 2s dwell 10 nN
applied force that has been scanned with contact mode AFM. After 1 hr of contact
with the substrate, it was found that contact mode would dislodge the higher part
of the features leaving toroidal structures. Panels C and F show zoomed in scans of
the remaining toroidal structures. ............................................................................. 78

Ryan P. Sullivan xiv

Figure 2-10: DPN deposited lines on PDA modified PDMS. The top images are
representative height map (A) and intensity map (B) of deposited lines. The profiles
C and D are transverse profiles of the thicker bottom line and thinner top line
respectively. As can be seen in B, the majority of the lines are highly reflective,
while tall deposits, particularly at the ends of the lines, are black. These black
regions correspond to the local peaks in the height map (A). Scale bars: 50 μm. .... 80

Figure 2-11: Optical Profilometry images and profiles of two lines. Images A, C, and
E are of a line printed at 0.5 μm/s while images B, D, and F are of a line printed at 5
μm/s. Panels A and B are height maps; C and D are optical images; E and F are
transverse height profiles. As can be seen the base material is highly reflective (C
and D), though deposited material above a certain height does not reduce and
appears black. This can be seen more clearly in the 0.5 μm/s line (panel C) where
the majority of the surface is black. It is unknown what causes certain lines to be
bisected in the middle. Scale bar: 10 μm. .................................................................. 81

Figure 2-12: AFM contact mode height maps of 50 μm/s samples that did, and did
not undergo plasma treatment. A) Samples without plasma treatment show a
morphology resembling a stack of many individual particles. We attribute this to
difference growth planes as the platinum would be pulled out of the ink and grow
from the substrate upwards; B) Samples treated with 10 min N2 plasma coalesced
into a single film where sufficient material was present (i.e. the rightmost part of
the line). Where there was not sufficient material, islands formed. C) Larger scale
image of PDMS-PDA substrate shows cracks. The blue arrows in B and C point to
cracks that were observed in the PDMS-PDA substrate and were attributed to the
plasma treatment. The cracks are brighter, rather than darker, because the edges
fold up, similar to a dried up lake (Zhang et al., 2013). Optical profilometry can have
difficulty with sharp valleys, detecting only the edges. Scale bars: (A) 5 μm; (B) 4
μm; (C) 10 μm............................................................................................................. 82

Figure 2-13: Optical images of SECM adverse sample reaction with redox mediator
solution. A) Deposition/reaction of SECM solution with PDMS-PDA substrate. B)
After 10 min IPA sonication; most of the material had been removed, but the
deposited platinum remained. Red arrows point to the line array of which
measurement was attempted. Scale bar: 500μm. .................................................... 84

Figure 2-14: Optical profilometry height map of features deposited using 10 s, 10

nN, 10 μm/s parameters of PDMS-PDA. During the 10 s contact time, the ink reacts
with the substrate. As the tip attempts to deposit subsequent drops, it interacts
with the previously deposited drop, causing it to merge into the existing meniscus.
This has the effect of moving a droplet along in discrete steps every 10 s or so

Ryan P. Sullivan xv
(including the withdraw, lateral movement, and approach between each dwell). As
can be clearly seen, what then remains are overlapping discs of material that have
reacted with the PDA surface. The large feature is the remaining ink left during the
final retraction............................................................................................................ 86

Figure 3-1: Layout of grid of m x n dots. In the ideal case 𝑥 and 𝑦 have the same
magnitude d and are orthogonal, i.e. θ = 90°. ........................................................... 92

Figure 3-2: Overview of protocol. .............................................................................. 96

Figure 3-3: Input Dialog Window. .............................................................................. 97

Figure 3-4: Results Window. The first seven columns contain data of a given
feature. Delta X values are formed with the neighbor on the right; Delta Y values are
formed with the neighbor below; Theta values are formed with both neighbors. In
the case where a neighbor doesn’t exist, i.e. the right most column or bottom row,
the value is left blank. Columns ten and eleven give cumulative statistical values for
the entire grid along with the parameters used in the processing and analysis. ...... 98

Figure 3-5: Graph of average inverse intensity along the x-axis of a grid on an 8-bit
scale before and after rolling parabaloid removal. ................................................. 100

Figure 3-6: Thresholding of Noise Samples. ............................................................ 101

Figure 3-7: Validation of Rolling Ball Subtraction Parameters. ............................... 102

Figure 3-8: Schematic of Insertion Sort Algorithm. ................................................. 104

Figure 3-9: Schematic of Grid Sort Algorithm. ......................................................... 105

Figure 3-10: Schematic of Nearest Neighbor Sort Algorithm. Using the top left
starting dot, the 8 ideal neighbor positions are calculated based on the input inter-
dot distance d. The nearest neighbors within 0.5-1.5 d are then matched to the
ideal positions, and are given a strength value based on how much they deviate
from that position. Found neighbors are then the seed for the next neighbor and so
on. ............................................................................................................................ 106

Figure 3-11: Optical images of inkjetted grids. Each row represents a set of specially
aligned grids. The top row uses parameters of 8 μs - 25 V, 3 kHz while the bottom
uses 8 μs – 30 V, 3 kHz. Printing was performed on a per line basis, e.g. the first row
in 1-6, followed by the second row in 1-6, etc. The images clearly show that while it

Ryan P. Sullivan xvi

is easy to obtain a single grid for publication, differences in parameters change the
viability of long term printing. While the 25 V waveform starts breaking down, the
30V waveform does not (this will be explored in detail in Chapters 4 and 5). ........ 109

Figure 3-12: Average theta values of each column in the two sets of grids seen in
Figure 3-11. .............................................................................................................. 110

Figure 4-1: Five phases of inkjet stimulation waveform.......................................... 133

Figure 4-2: Schematic representation of C-AFM measurements. ........................... 137

Figure 4-3: Representative Images of droplets (10 μs pulse) at various voltages (22-
27 V) with a head traveling downwards. ................................................................. 138

Figure 4-4: Schematic of a droplet with a tail. A given slice (black), has a thickness of
xk and a diameter of yk ............................................................................................. 138

Figure 4-5: Velocities of the droplets as a function of the applied voltage (V) and
pulse length (μs). ...................................................................................................... 141

Figure 4-6: Distance until sphere is the distance the droplet needs to travel before
forming a perfect sphere. ........................................................................................ 142

Figure 4-7: Representative optical images of unreduced inkjetted features on PDMS

that represent the four main phases of jetting optimization. ................................. 144

Figure 4-9: Optical images of inkjetted grids. .......................................................... 149

Figure 4-10: Optical (A–C) and SEM (D-F) images of deposited features. (A)
Unreduced ink immediately after deposition; (B) unreduced ink immediately after 1
hr under vacuum exhibits no spreading; (C) after 10 min plasma reduction. The
presence of oxygen species create hydroxyl groups on the surface of the PDMS,
increasing its hydrophilicity and facilitating spreading of the ink (Bhattacharya et al.,
2005). (D) Thermally treated (200C) samples exhibit no spreading; (E) Plasma
treated samples on PDMS exhibit spread similar to C; (F) Plasma treated samples on
Au-Mylar do not exhibit spread, further supporting surface modification as the
cause of spreading. Scale bars: 20 μm (A-C), 100 μm (D-F). ................................... 152

Figure 4-11: Pictures of Pt-PDMS samples on glass slides and dishes in the plasma
chamber during treatment. A: EG atmosphere created by addition of mL droplets of

Ryan P. Sullivan xvii

EG; B: Air; C: The chamber was pumped down to 500 mTorr and back up to 1600
mTorr 5 times before finally resting at 1050 mTorr; D: post 30 min N 2. ................. 153

Figure 4-12: Areas of six features to measure evaporation of a 22 min interval after
printing. Measurements were made roughly every 1 min. .................................... 154

Figure 4-13: Reduced and unreduced sides of a 0.5 μL dropcast 10% Pt ink on
atmospheric plasma treated 1 x 0.5 cm glass pieces. The top side, in contact with
plasma, is reduced and highly reflective. The bottom side, as seen through the glass
substrate, is brown/black and non-reflective. ......................................................... 155

Figure 4-14: Optical images of progression of reduction of 0.5 μL dropcast platinum

ink on non- plasma treated glass over repeated N2 plasma treatments. Duration
listed is total duration. A) 20 min total plasma treatment produced mostly black
material with metallic edges. B) 40 min total plasma treatment caused islands of
black material to recede, increasing the size metallic edges. C) 60 min total plasma
treatment caused most of the material to reduce. D) 80 min total plasma treatment
completely reduced the top sample while leaving a small island of partially reduced
material in the center of the bottom material. E) H2O washed sample using flash
photography shows the high reflectivity of the reduced Pt. ................................... 156

Figure 4-15: Representative C-AFM images of reduced features on Au-Mylar. ..... 157

Figure 4-16: Representative C-AFM IV plots............................................................ 158

Figure 4-17: Plot of average total system resistance obtained from averages of C-
AFM IV curves at various set points. Error bars represent ±1 standard deviation. 160
Based on the statistical variance, further studies may need an order of magnitude
higher population size to find a statistically significant difference for a single applied
force, and two orders of magnitude higher population size to find statistically
significance differences for all applied forces to account for changes in contact area.
....................................................................................... Error! Bookmark not defined.

Figure 4-18: Lowvac SEM EDS Maps of a Pt feature on PDMS ................................ 162

Figure 4-19: Optical image of ink jet head. .............................................................. 164

Figure 4-S1: Graph of error in area (light blue shading) as a percentage of the true
area (dark blue line) as a result of single a pixel difference on thresholded edges. In
the case of the spherical geometry of the droplets cross-sections, the diameter
changes ±2 as they have two edges. ........................................................................ 170

Ryan P. Sullivan xviii

Figure 5-1: Velocities of 1 pL droplets of 10% Pt weight ink as a function of the
applied voltage (V) and pulse length (μs). ............................................................... 179

Figure 5-2: Distance until coalescence of 1 pL droplets of 10% Pt weight ink as a

function of the applied voltage (V) and pulse length (μs). ...................................... 180

Figure 5-3: A and C are pattern schematics for the calibration grids shown in optical
images B and D respectively. ................................................................................... 181

Figure 5-5: Photograph of reduced printed platinum features on PDMS-PDA

substrates with ruler for scale. ................................................................................ 187

Figure 5-6: A and C are schematics of the inkjet patterns used to print the lines
shown in B and D respectively. ................................................................................ 188

Figure 5-7: AFM Images of PDMS-PDA surface (A, B), PDMS-PDA having undergone
10 min N2 plasma treatment (C); Reduced Pt on PDMS-PDA after 10 min N2 plasma
treatment (D). Panel A shows the homogenous nature of the PDA film, while the
zoomed in image B shows the nanoporous nature of the PDA film, agreeing with
images in literature .................................................................................................. 191

Figure 5-S1: A calibration image of spheres creating using GIMP (GNU Image
Manipulation Program) with diameters from 1 – 139 pixels with a hardness of 1.
The calibration image was fed into ImageJ to determine the effect of pixel size of a
perfect circle on the circularity. ............................................................................... 195

Figure 5-S2: Circularity of circles from calibration image (Figure 5-S1) as a function
of diameter in pixels................................................................................................. 196

Figure 5-S3: Image of Cochlear implant................................................................... 196

Ryan P. Sullivan xix

 LIST OF TABLES

Table 1-1: Survey of metal salt and reducing agent combinations appearing in
literature. ................................................................................................................... 52

Table 1-2: Plasma reduction of metal salts................................................................ 55

Table 2-1: Surface roughness values on unmodified and PDA modified (1 hr) PDMS
substrates. The top two rows are at x50 magnification while the bottom two rows
are at x100 magnification. See Section 2.5 Appendix for a full explanation of how
the values were calculated. ....................................................................................... 72

Table 3-1: Validation of Rolling Ball Subtraction. .................................................... 101

Table 4-1: Metal loading lookup table for ink formulations based on desired Pt
content. .................................................................................................................... 132

Table 4-2: Average volume data obtained by summating circular slices

perpendicular to the travel vector of droplets in flight with 3 individual ink jets. Lack
of standard deviations indicates that only a single jet produced quantifiable data.
.................................................................................................................................. 139

Table 4-3: Diameter, area, and volume averages of features using jets at 3 kHz firing
rate. Distance and volume values were derived from the area data using the
roundness and contact angle relationships. ............................................................ 146

Table 4-4: Average theta values of grids using jets at 3 kHz firing rate. Theta is the
angle formed between a given drop and two neighbors; in a perfectly placed grid,
all theta values would be 90°. Deviation from 90° therefore indicates lower
reproducibility in droplet placement. ...................................................................... 147

Table 4-5: Average circularity and roundness of printed features using jets at 3 kHz
firing rate. ................................................................................................................. 148

Table 4-7: Average width and height of features after the different treatments
represented as a percentage of before treatment width and height. .................... 150

Ryan P. Sullivan xx
Table 4-8: Average EDS values of 7 features per parameter, taken using Low-Vac
SEM of 10 pL deposited 10% Pt weight inks on PDMS and Au-Mylar. Cl ratios are
highlighted in red. Complete reduction should have Cl ratio of 0:1 while incomplete
reduction should have a Cl ratio of 6:1. ................................................................... 162

Table 5-1: Average volume data (pL) obtained by summating circular slices
perpendicular to the travel vector of droplets in flight with ................................... 178

Table 5-2: Geometric values of printed drops obtained from the ImageJ plugin. .. 183

Ryan P. Sullivan xxi

 LIST OF ABBREVIATIONS

CNS: Central Nervous System

CNT: Carbon Nanotube
CVD: Chemical Vapor Deposition
DNA: Deoxyribonucleic acid
DPN: Dip-Pen Nanolithography
ECM: Extracellular Matrix
ECoG: Electrocorticography
EDS: Elemental Detection System
EEG: Electroencephalography
EMG: Electromyography
MEA: Multielectrode Array
NGF: Neural Growth Factor
NP: Nanoparticle
PDA: Polydopamine
PDMS: Polydimethylsiloxane
PEDOT: Poly(3,4-ethylenedioxythiophene); a conductive polymer
PSS: Polystyrene Sulfonate
PNS: Peripheral Nervous System
PPy: Poly(pyrrole); a conductive polymer
PVP: Polyvinylpyrrolidone
RGD: L-Argenine-Glycine-L-aspartic acid tripeptide
RIE: Reactive Ion Etching
SEM: Scanning Electron Microscopy
SIT: Super Inkjet Technology
TBI: Traumatic Brain Injury

Ryan P. Sullivan xxii

 ABSTRACT

Sometime in the 1790’s an Italian physician by the name of Luigi Alosio Galvani was
working with a frog preparation when one of his colleagues touched the sciatic
nerve with a statically charged scalpel. The resulting spark caused the frog leg to
contract violently, seemingly springing to life. It was this discovery that gave birth to
the field of electrophysiology and all subsequent fields which build upon it. Because
of this discovery and continued research with “animal electricity” in muscle and
neural tissue, Galvani lends his name to the galvanostat, an important device used
in electrochemistry to apply constant current despite varying loads.

While various new technologies for stimulating and recording tissues have
developed since the time of Galvani, metal electrodes have remained the
quintessential tool of neuroscience, with the famous experiment becoming a
mainstay in basic scientific education. Metal electrodes are not only research tools
but have been expanded into the clinical setting for use in chronic implants. Flexible
electrode arrays have allowed for the successful development of commercially
available implants, such as the cochlear and retinal implants, that can restore
hearing and sight respectively. However, traditional fabrication methods are limited
for the soft, flexible, and biocompatible materials which are required to increase
the fidelity of these implants. Additionally, the high cost of production has always
been an issue for the commercial viability of medical devices. For these reasons,
additive manufacturing technologies hold significant promise for a higher degree of
custom designed implants on a per patient basis, while reducing costs due to the
elimination of the masks and/or etching steps used in current manufacturing
techniques.

This thesis therefore aimed to contribute to the ever growing body of knowledge
working to solve the issues associated with printing platinum on PDMS, by
extending our previous work on polyol inks for use with DPN on flexible substrates.
This thesis also examined methods for transitioning from grid to line printing with

Ryan P. Sullivan xxiii

DPN. We have shown for the first time that platinum inks can be used in inkjet
printing for high speed deposition at the micron scale; and have developed a new
rapid analytical method that allows for a high degree of validation when optimizing
jetting parameters. We have used both 10 pL and 1 pL inkjets to demonstrate that
grids of micron scale platinum features can be deposited at kHz speeds. Finally we
explored methods for transitioning from printing grids to lines using inkjet printing
to fabricate centimeter scale platinum lines.

In translating DPN from silicon to PDMS we found that approach and withdraw
speed are extremely important. Slow speeds (1 μm/s) were found to allow the ink
to pull back off the substrate onto the tip during withdraw. High speeds (100 μm/s)
were able to deposit material, however this also punctured the substrates.
Intermediate speeds of 10 μm/s were able to reproducibly deposit features onto
the substrates. Polydopamine modification of the substrates allowed for the
deposition of 90 μm long platinum lines through reactive printing.

Poor internal optics of the inkjet printer necessitated the development of software
to allow for analysis to be performed on a high number of features (roughly half a
million in total) using normal microscopy equipment. An ImageJ plugin was created
which is able to automatically detect and analyze optical images of calibration grids.
The high number of data points obtained with this software allowed for the
differentiation of closely overlapping feature distributions with p-values from
2.04x10-9 to 6.12x10-44, allowing for the high certainty which is needed for large
scale fabrication.

Using the abovementioned software, inkjet waveforms were developed for 10 and
1 pL ink cartridges, allowing for the rapid deposition of platinum inks between 1
and 10 kHz. The time interval between the deposition of the ink onto the substrate
and plasma reduction of the ink was found to be vital, as additional time was found
to impede the reduction process and floors the conductivity of the printed features.
Due to the hydrophobicity of PDMS, line printing with inkjet was similarly found to
be difficult, with the ink from the printed lines separating into discrete spheres on

Ryan P. Sullivan xxiv

the substrate. Polydopamine modification was again found to be helpful, and
allowed for the deposition of 175 μm wide lines at rates of 75 mm/s.

Overall the work contained in this thesis has contributed to overcoming the issues
of printing platinum inks on PDMS, providing another tool to be used towards an all
in one additive fabrication device that can create custom neural implants consisting
of PDMS, platinum, and cells.

Ryan P. Sullivan xxv

 ACKNOWLEDGEMENTS

I am deeply grateful for all the time, money, and work my supervisors have spent training
and aiding me during my candidature. I met many hurdles during my PhD and my
supervisors were vital in overcoming them. I’d like to thank Zhilian for her advice on PDMS,
PDA, and plasma treatment. Her ideas formed the basis for the original plasma reduction
parameters as well the use of PDA as an adhesion layer. I remember on my first day in
Australia, after having arrived early and not knowing a single person, Zhilian came and took
me grocery shopping and helped me set up. She provided a daily friendly face and anchor.
I’d like to thank Michael, for his expertise on all matters DPN and AFM, for taking me on
after my funding and project completely changed, and for helping me continue the DPN
work on the Asylum AFM after the dedicated DPN broke and NanoInk went under. I’d lastly
like to thank Gordon Wallace for constantly fighting for and supporting me during my
candidature and for always thinking about the research and students first. This PhD would
not have been possible without all of your help.

To all my fellow IPRI students and researchers, thank you for being such a warm and
supportive community. I need to particularly thank Cathal O’Connell, who trained me on
the DPN and whose work laid the foundation for this thesis as well as Binbin Zhang for
training me on the inkjet printer and providing moral support. I’d additionally like to thank
Joseph Giorgio for acting as constant support and close friend, off whom I could always
bounce ideas.

To the EMC staff, thank you for your expertise and support. I want to particularly thank
Tony Romeo for training me on AFM, SEM, EDS, and microscopy sample preparation, and
for helping me move into my new apartment. I’d additionally like to thank Mitchell
Nancarrow for training me on lowvac SEM and EDS and taking a keen interest to my work.

To my parents, I am eternally grateful for their warm wishes and financial support. My
academic journey started well before this PhD, and it is due in part to them fostering my
scientific interests that I am at this point. To Evelyn Andrews, thank you for putting up with
me and providing me with a home while I wrote this thesis.

And of course this thesis would not have been possible without the love of my life, Dr.
Jessica Andrews. Without your input and constant emotional and financial support while
writing, this thesis this would not have been possible.

Ryan P. Sullivan xxvi

 CHAPTER ONE: INTRODUCTION AND
REVIEW OF THE LITERATURE

1.1 INTRODUCTION TO NEURAL PROSTHETICS

1.1.1 History of Prosthetics

Prosthetics are devices which serve to replace lost or damaged appendages and

organs in order to regain or enhance lost function. The first documented

prosthetics were artificial toes for partially amputated feet, dating all the way back

to ancient Egypt, between 950 and 710 BCE (Figure 1-1) (Finch, 2011; Finch et al.,

2012). Modern replication and analysis of these prostheses has demonstrated

improved performance and comfort for patients, indicating actual medical benefit

beyond just aesthetics (Finch et al., 2012). Development of prostheses would

continue throughout the centuries, with particularly notable examples being the

iconic peg-leg and hook, often seen in popular pirate literature, and intricate

Victorian metal arm prosthetics, popular in the modern steampunk culture. In the

1500’s the infamous Gottfried von Berlichingen, a knight in the Holy Roman Empire,

lost his arm to cannon fire at the age of 24; he had two prosthetic arms made for

him, which allowed him to continue fighting for a further 40 years, earning him the

name Götz of the Iron Hand (Figure 1-1) (Mechel, 1815).

Ryan P. Sullivan 1
Figure 1-1: Ancient Prosthetic devices. An ancient false toe from an Egyptian mummy (Finch, 2011),
and drawings of the iron hands of Gottfried von Berlichingen (Mechel, 1815). Reprinted from Finch,
J., (2011) The ancient origins of prosthetic medicine; The Lancet; 377(9765), 548-549, with
permission from Elsevier. Copyright © 2011 Published by Elsevier Ltd.

Jumping forward to the 1950s, great advances in prostheses were afforded by the

development of bioinert materials (Hedrick et al., 2006). This was the last piece

needed for the development of implanted prosthetics. While previous implants

could only be strapped or mounted to the outside of one’s body, these newly

developed materials allowed for the implantation of permanent prosthetics.

One particular type of prostheses are neuroprothetics, which interface with the

human nervous system. Neural interfacing allows for a higher degree of control of

the prostheses, well beyond that of electromyograms (EMGs), as well as also being

able to restore certain sensation (Borton et al., 2013; Formaggio et al., 2013). The

cochlear implant is the oldest commercial neuroprosthetic, with the first

implantation surgery occurring in 1978 (Figure 1-2) (Clark et al., 1979). In the past

10 years, 213,909 cochlear devices have been implanted worldwide, not accounting

for devices produced by other companies. In this past year alone an average of 74

cochlear implantations occurred each day (Cochlear Ltd., 2016).

Ryan P. Sullivan 2
Figure 1-2: The first cochlear implant. A photograph of the UMDOLEE multiple electrode prosthesis
for implantation next to a 20 cent piece. Reprinted from Clark GM, Pyman BC, Bailey QR., The
surgery for multiple-electrode cochlear implantations. J Laryngol Otol. 1979 Mar;93(3):215-23 with
permission from Cambridge University Press.

1.1.2 Neural Interfacing Techniques

There are two fundamental interactions which occur between a neural interface

and target neural tissue in terms of signaling: stimulation, i.e. sensory input, and

recording, i.e. motor output. Prosthetics such as artificial arms and legs are

primarily concerned with recording motor output from the nervous system to

govern their movement as close to the original appendage as possible. Prosthetics

such as retinal implants, e.g. Argus II (Second Sight, USA), and cochlear implants,

e.g. Cochlear Nucleus (Cochlear Ltd., Australia), on the other hand are primarily

concerned with stimulation to restore sensations of sight and hearing respectively.

As prosthetic technology improves however, both stimulation and recording are

becoming necessary to fully reproduce functionality. In the case of artificial arms for

example, stimulation becomes important for restoring proprioception,

Ryan P. Sullivan 3
thermoception, nociception (in terms of damage to the prosthetic) and

mechanoreception, which are vital components that are integrated into complex

movement (Tabot et al., 2015). In retinal implants, recording could be used to

control aspects of the device, such as variable magnification of intra-ocular lenses

(Conrad, 2016) or Heads-Up Displays (HUD) for information overlay, which is of

particular interest for military applications.

Electromyography (EMG) is one of the earliest recording techniques successfully

used to control prosthetic arms. Surface EMG is a noninvasive technique that works

by placing electrodes on the skin to measure aggregate action potentials of muscle.

The aggregate action potential of muscle is considerably larger than that of nerves,

making it easier to measure. The downside however is that prosthetics using

surface EMG as the source signal are relatively clumsy and crude. Patients must

learn to repurpose their existing muscles to send gross signals to the prosthetic to

move. The prosthetics are limited to predefined finite-states such as grasping and

pointing (Figure 1-3). This means instead of seamless and arbitrary enervation to

hand, the patient must focus on choosing a particular action from a list of

predefined actions. Because of this limited command space, buttons are often used

to change the meaning of a particular contraction (Tabot et al., 2015). Surface EMG

is effectively the low hanging fruit of all techniques, but is incapable of ever

producing the seamless movement and sensation of an original appendage.

Ryan P. Sullivan 4
Figure 1-3: Prosthetic hand in eight finite-states. Reprinted with permission from Dalley, S.A.,
Bennet, D.A., Alshammary N.A., Goldfarb M. Multigrasp Hand Prosthesis and Myoelectric Control
Method for Enhancing the Functional Capability of Upper Extremity Amputees; 2012; IEEE Life
Sciences; Copyright © 2016, IEEE.

Electroencephalography (EEG) is a recording technique that works by placing a

nerve cap over the skull (above the skin) (Figure 1-4) to obtain a large area of

potentials in the brain, though it has relatively low resolution. It is non-invasive by

nature and is used to derive intent-related inputs (Borton et al., 2013). Similar to

EMG, patients must learn to control prosthetic devices by focusing on the intent of

an action. For example, a patient will focus on grasping with the lost hand multiple

times while EEG is recording to learn the patient’s particular pattern during that

intent. The device will then respond, coupling that firing signature with the specific

finite-state (Figure 1-3). The technique is more versatile than EMG as intent states

may be learned for any given action, irrespective to how much of an appendage the

prosthetic replaces.

Ryan P. Sullivan 5
Figure 1-4: Three types of recording devices for neural interfacing; electroencephalography (EEG),
electrocorticography (ECoG), and microelectrode array (MEA). EEG consists of many electrodes
placed along the scalp or neck. ECoG consists of an electrode array placed on the surface of the
brain. MEAs penetrate the outer brain tissue, allowing them to have more localized recording than
either EEG or ECoG.

Electrocorticography (ECoG), also known as intracranial electroencephalography

(iEEG), is an invasive technique that places electrodes directly on the surface of the

brain (Figure 1-4). To perform this, a craniotomy and resection of the dura is

required (Murphy et al., 2016). It does however provide much stronger signals than

EEG, having less insulating material between the electrodes and target tissues.

Ryan P. Sullivan 6
Furthermore, each patient will have differing recording fields and electrode

positions due to individual anatomies (Formaggio et al., 2013).

Multielectrode arrays (MEA) are by far the most invasive of the aforementioned

techniques but provide the largest temporospatial resolution due to their localized

nature. As the name suggests, MEAs are an array of multiple electrodes that are

inserted directly into the target tissue, allowing for high specificity (Figure 1-4).

Additionally unlike the other three techniques mentioned above, MEAs can also

stimulate target neural tissue. They can have resolutions down to millisecond time

scales, along with sub-cellular resolution, and the ability to localize individual cells

(Obien et al., 2015).

Finally, optogenetic stimulation is a relatively new technique that uses light pulses

to stimulate and silence firing of targeted neural tissue. It works by introducing light

gated channel proteins, particularly of the rhodopsin family, into axonal

membranes (Bass et al., 2010). The benefits of this technique include high

specificity, allowing for the stimulation of individual neurons, and removal of cross

talk when used in concert with recording electrodes (Bass et al., 2010). In a setup

where electrodes are used for both stimulation and recording, the stimulation

signals must be removed from the recorded signals; optogenetic stimulation is

capable of removing this obstacle. However, the ethics surrounding the

implantation of optogenetic devices are far more severe than those of MEA’s. While

both implants require surgery, optogenetic stimulation requires the introduction of

opsin channel proteins, usually through retroviral vectors, though other techniques

Ryan P. Sullivan 7
such of electroporation have been used as well (Erofeev et al., 2015, Optogenetics:

A Roadmap 2018). Due to the hurdles of ensuring this approach is safe and ethical,

this technique may not reach clinical applications for decades. For a more in depth

review on the use of optogenetics for prosthetics see Jarvis and Schultz (2015).

All of this considered, MEA’s are the most widely used of the aforementioned

neural interfacing approaches for advanced prosthetics. Different types of MEAs

along with commercial examples are detailed in Section 1.2.

1.2 TYPES OF NEURAL ELECTRODES AND LOCATIONS SITES

While all MEAs consist minimally of stimulating electrodes with an insulating cover,

the location of an implant has strong effects on the component materials, the 3D

design, and the surgical technique, along with the encountered immune reactions

(Grill et al., 2009). In general, locations can be divided into 4 broad categories: the

brain, the spinal cord, peripheral nervous system (PNS), and central nervous system

(CNS) adjacent systems such as the eye and cochlea. Each location has its benefits

and drawbacks, primarily in device efficacy, complexity, and invasiveness (Grill et

al., 2009).

1.2.1 Hearing

Hearing works by focusing sound waves, i.e. pressure waveforms, from external

media to the ear drum. The three tiny bones in the middle ear, known as ossicles,

then transfer vibrations from the ear drum to the oval window, the interface of the

liquid filled cochlea (Figure 1-5). The basilar membrane inside the cochlear is

Ryan P. Sullivan 8
tonotopic, with specific locations having specific resonance frequencies. Due to the

three dimensional structure of the cochlea, high frequencies are detected at the

basal end while low frequencies are detected at the apex. Along the basilar

membrane are hair cells that act as mechanoreceptors, which become depolarized

when the amplitude of their characteristic frequency reaches certain thresholds

(Figure 1-5). This causes them to release neurotransmitters at synaptic clefts of the

cochlear nerve, and allows for the transduction of nerve impulses along the

cochlear nerve (Figure 1-5 and Figure 1-6).

Figure 1-5: Schematic representation of sound transduction and anatomy of the cochlea. Sound
waves focused from the outer ear travel through the ear canal to the tympanic membrane. The
perturbations are then transferred through the three ossicles, the malleus, incus, and stapes, to the
oval window.

Encoded signals are transmitted along this cochlear nerve to the cochlear nucleus in

the brainstem (Figure 1-6). These nerve impulses are subsequently transmitted to

Ryan P. Sullivan 9
the inferior colliculus where auditory signals are integrated with other signals for

subconscious reflexes. The inferior colliculus in turn projects to the medial

geniculate nucleus of the thalamus, which itself projects to the primary auditory

cortex where conscious awareness of sound is achieved (Figure 1-6).

Figure 1-6: Schematic representation of nerve impulse transduction pathway to the auditory cortex.
Hair cells in the cochlea stimulate the cochlear nerve, which projects to the cochlear nuclei. From
here, signals propagate to the Inferior Colliculus, the medial geniculate nucleus, and finally the
primary auditory cortex.

Currently, auditory prosthetics target one of four locations along this path. The

most common and least invasive prosthetic is a hearing aid, which serves to amplify

Ryan P. Sullivan 10
the sound waves reaching the ear drum. This however requires that the entire

auditory system be intact.

In cases where there is damage to the middle ear, including the ear drum and

ossicles (Figure 1-5), cochlear implants may be used. First developed in 1978,

cochlear implants are electrode arrays implanted directly into the cochlea (Brill et

al., 2009; Clark et al., 1979). Instead of amplifying the pressure waves to amplify the

signal from the depolarized hair cells, the electrode array seeks to bypass the hair

cells altogether by directly stimulating the cochlear nerve with electric fields (Brill et

al., 2009; McCreery et al., 2000; Rebscher et al., 2008). By having many individual

electrodes along the length of the basilar membrane of the cochlear, stimulation

can mimic the tonotopic response of the hair cells, though to a lesser degree.

The next most invasive implant is the auditory brainstem implant. Instead of

stimulating the cochlear nerve (which projects directly to the brainstem), this

implant directly stimulates the brainstem (Figure 1-6) (Schwartz et al., 2008;

Sennaroglu and Ziyal, 2012). This is needed in cases where there is trauma or

developmental defects to the cochlea, rendering the previous two implant types

useless. This includes disorders such as neurofibromatosis type II (NF2), auditory

nerve aplasia, and cochlea ossification (Colletti et al., 2012, 2005; Schwartz, 2014;

Schwartz et al., 2003). The first auditory brainstem implant was implanted in 1979

(House and Hitselberger, 2001).

The final and most invasive implant for hearing is a cortical implant directly

targeting the auditory cortex (Figure 1-6). Limited research on these cortical

Ryan P. Sullivan 11
implants has been done in rodent studies, but due to the invasive nature of the

implant as well as the increased complexity of the neural encoding required,

according to the literature no clinical work has yet been attempted (House and

Hitselberger, 2001). At each step along the pathway the signals representing sound

undergo further processing, changing the encoding away from the simple tonotopic

mapping of the cochlea.

1.2.2 Vision

Vision works by detecting light with specialized photoreceptors in the eyes. Light

enters the eye anteriorly, passing through the cornea, pupil, and lens; this

combination acts as a compound lens, refracting light onto the retina on the

posterior wall of the eye (Figure 1-7) (Yue et al., 2016).

Figure 1-7: Schematic representation of a cross–section of an eye. Light travels through the cornea,
pupil, lens and projects onto the back of the retina. The retina (bottom panel) contains
photoreceptors for color vision (cones) and low light, or black and white vision (rods). There are 3

Ryan P. Sullivan 12
sites for visual implants within the retinal layers: epi-retina, sub-retina, and supra-choroid (top
panel).

The retina contains two types of photoreceptors responsible for conscious vision,

rods and cones (Figure 1-7). The three types of cones cells, short 425nm (Blue),

medium 530nm (Green), and long 522-557nm (Red), are primarily responsible for

color vision, while rod cells are primarily responsible for scotopic vision, the

monochromatic vision in low light environments. All of these receptors function by

containing different opsins, proteins which change conformation when they absorb

a sufficient amount of photons, with unique absorption spectra. This

conformational change induces hyperpolarization of the photoreceptor. Arrays of

photoreceptors propagate to bipolar and retina amacrine cells, which in turn

propagate to retinal ganglion cells (Figure 1-7). The collection of axons from these

ganglia forms the optic nerve leading to the optic tract and lateral geniculate

nucleus (LGN) in the thalamus (Figure 1-8), and to the superior colliculus on the

brainstem. The LGN deals with processing of the neural signals, while the superior

colliculus integrates the signals for motor control of the eyes, in addition to

responding to visual stimuli. The LGN subsequently projects to the primary visual

cortex, where the bulk of visual processing occurs (Figure 1-8).

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Figure 1-8: Schematic representation of the neural pathways for vision. From the retina, neural
signals for vision travel along the optic nerve, the optic tract, synapse on the lateral geniculate
nucleus in the thalamus and then project to the visual cortex. The different sites for the 5 types of
visual implants or stimuli are marked.

Similar to hearing, the choice of visual implant begins with the least invasive site, in

this case the eye, and works along the neuronal path, with each subsequent site

increasing in signal filtering and invasiveness.

The most common case of visual impairment is where the focusing system is

damaged, but from the retina onwards is functioning. Often external lenses, i.e.

Ryan P. Sullivan 14
glasses or contacts, can be a used to correct the optics. There has been recent

interest in intra-ocular implants that alter the focusing system itself, allowing the

possibly of actively controlled optic power (Conrad, 2016).

Retinal implants are used when the photoreceptors are nonfunctional but the

intermediate and ganglion cells are functional. This can be caused by diseases such

as Retinitis pigmentosa, which affects around 1.5 million people (den Hollander et

al., 1999), age related macular degeneration, which affects over 2 million people in

the United States, and 30-50 million people worldwide (Bressler, 2004; Friedman et

al., 2004), and choroideremia. Retinal implants function by using electrode arrays in

a grid pattern to stimulate the intermediate and ganglion cells of the retina, with

each electrode corresponding to a different group of photoreceptors, seeking to

replace their functionality (Yue et al., 2016). Higher electrode densities more

accurately map the original photoreceptor activation. The retinal ganglion has a

range of densities from 1750 to 31000 neurons per square millimeter depending on

eccentricity towards the periphery, meaning that a similar density of electrode

stimulation sites would be required at the minimum to restore complete vision

(Harman et al., 2000). There are three different types of retinal implants: epi-

retinal, sub-retinal and supra-choroidal. As seen in Figure 1-7, epi-retinal implants

are placed on the surface of the retina, sub-retinal implants are placed between the

retinal pigment epithelium and the outer retinal layer (Weiland et al., 2005), while

supra-choroidal implants are placed between the choroid, which is the blood vessel

network of the eye at the back of the retina and the sclera (Yue et al., 2016).

Ryan P. Sullivan 15
The first retinal implant was used in 1997 (Dawson and Radtke, 1977), and since

then, prosthetics for all three sites have undergone clinical trials (Ayton et al.,

2014). Argus II, an epi-retinal implant produced by Second Sight Medical Products,

received approval for treatment of retina pigmentosa in Feb 2011 in Europe, and

FDA approval to market their device under a Humanitarian Device Exemption Feb

2013 in the US (U.S. Food and Drug Administration, 2013). The Argus II is a grid of

6x10 Platinum Gray electrodes, which is well below the theoretical maximum

needed, though future versions may increase this number (Weiland and Humayun,

2014). The Argus II also represents a x3 increase in channel number, and a x10

density of electrodes compared to cochlear implants (Weiland and Humayun,

2014). A second retinal implant, the Alpha-IMS, made by Retina Implant AG in

Germany, also received approval in Europe in 2013 (Chuang et al., 2014). For

further information on retinal implants, see the latest review by Yue et. al. (2016).

In cases where the retina is damaged, implants may be used to stimulate the optic

nerve directly (Figure 1-8) (Margalit et al., 2002; Veraart et al., 2003). The surgery is

more invasive, and techniques are still under development (Sakaguchi et al., 2012).

The electrode designs that have been tested thus far have been multiple wires or

cuff electrodes to wrap around the optic nerve (Delbeke et al., 2002).

The third approach to restoring vision is to interface with the visual cortex directly

(Figure 1-8). While cortical implants will function regardless of the status of the

retina and the optic nerve, allowing it to be potentially used in more patients, like

the auditory cortical implants from section 1.2.1 above, the surgery is invasive and

Ryan P. Sullivan 16
the signaling requirements are much more complex. While the retina and the optic

nerve, which is just the axonal projections of the retinal ganglia as mentioned

above, are rather straight forward in their neural encoding of vision, the cortex is

much more complicated and segregated (Fernandes et al., 2012). Different visual

processing tasks occur at different locations of the cortex, making it more difficult

to project a full image compared to a retinal electrode array. The first cortical

prosthesis for vision was in 1968 (Brindley and Lewin, 1968).

1.2.3 Arms and Legs

While goals for movement originate primarily in the dorsolateral prefrontal cortex,

actual movement signals originate in the motor cortex (Figure 1-9A). The main

focus of implants for prosthetic appendages is therefore recording efferent signals

originating in the motor cortex, to be transmitted by the corticospinal pathways

(Figure 1-9A). Any goal-driven implant will be fundamentally limited and not reach

seamless use. Additionally, feedback from the prosthetic device, particularly

proprioception and mechanoception (transmitted by the dorsal columns-medial

lemniscus pathway; Figure 1-9B), is of vital importance for complete restoration of

movement. Many prosthetic arms currently require visual monitoring by the patient

to know its position and have separate processing to control grip strength. Between

40-60% of patients choose not to use prosthetic arms because of these difficulties,

and cite lack of sensory feedback as a major factor (Biddiss and Chau, 2007a, 2007b;

Biddiss et al., 2007; Resnik et al., 2012).

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Figure 1-9: A. Corticospinal tracts – descending pathway originating in the motor cortex, responsible
for motor control of the proximal and distal arms and legs. B. Dorsal columns-medial lemniscus
pathway – ascending pathway originating in the limbs, responsible for transmitting discriminative
touch and proprioception feedback from the proximal and distal arms and legs. (Reprinted from
TeachMeAnatomy.com (http://teachmeanatomy.info/neuro/pathways/ascending-tracts-sensory/
and http://teachmeanatomy.info/neuro/pathways/descending-tracts-motor/) © 2015-2016
TeachMeAnatomy.com, under the Creative Commons Attribution-NonCommercial-No Derivatives
4.0 International license (https://creativecommons.org/licenses/by-nc-nd/4.0/).

As previously mentioned myoelectric prostheses use electromyography (EMG) and

may be used to record potentials from contracting muscles at the remaining end of

a missing appendage (González-Fernández, 2014). This however is limited to

recording only and works in an ad hoc manner, requiring patients to learn to move

the remaining muscles in a non-normal manner to control the prosthetic.

A more accurate, though difficult approach is to interface the prosthesis with the

remaining efferent and afferent neurons in the appendage; however, the main

issue here is to overcome Wallerian degeneration of the remaining neurons (Grill et

Ryan P. Sullivan 18
al., 2009). When an axon succumbs to trauma, the entire distal segment will break

down and be phagocytosed. The distal end of the proximal segment will attempt to

regrow to the to the original innervation site if the neurolemma is intact. As such,

growth factors and cell populations introduced with the electrode interface become

important to coaxing growth onto the electrode surfaces (Grill et al., 2009; Tan et

al., 2013).

1.2.4 Memory

Unlike vision or hearing which have clear origin and end points, as well as clear

discrete locations for implants, memory is much more complex. There are many

types of memory, and many other systems which are integrated into forming,

storing, and retrieving memory which are far beyond the scope of this literature

review. It is clear however, that the hippocampus plays a large role in memory

formation, and is therefore a target for neural implants. Since 2000, over 270,000

US military service members and 1.7 million US civilians have suffered from

Traumatic Brain Injury (TBI), impairing memory (Faul et al., 2010); roughly one

fourth of all combat deployed US personnel will suffer a TBI from explosive devices

(Heinzelmann et al., 2014). These figures have increased interest in the

development of methods for restoring memory function. Recent work in rodent

studies has shown that a prosthetic device can restore and enhance memory

storage by targeting the CA3 and CA1 regions of the hippocampus (Figure 1-10)

(Berger et al., 2011; Gilbert et al., 2015; Hampson et al., 2012; Song et al., 2015; Xu

et al., 2015). On August 16, 2016 it was announced that a new company, Kernel,

Ryan P. Sullivan 19
had been formed to take the work of Ted Berger to a clinical setting with the

development of human memory implants (Strickland, 2016).

Figure 1-10: Schematic of Hippocampal implants interfacing between CA3 and CA1. Due to lateral
symmetry, there are two mirrored hippocampi, and therefore two mirrored implants.

1.3 ELECTRODE MATERIALS AND DESIGN

1.3.1 Overview of Implant Challenges

Neural electrode design varies depending on the location of the target site. The

encountered immune response, the 3-dimensional structure of the implant, as well

as mechanical differences of the tissues are all region specific. That said, there are

similarities between sites. The CNS immune response and mechanical needs are

identical for implants in the visual, auditory, and motor cortices, the auditory

brainstem, as well as the hippocampi (Grill et al., 2009). Retinal and cochlear

implants are similar in structural needs, to the point that the Argus I & II are based

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directly on the Advanced Bionics cochlear implant technology. Spinal implants and

conduits have their own issues. Lastly, peripheral implants for arms and legs may be

relatively distant to one another but share the same immune response and

mechanical needs.

While parameters for these four groups of implants may differ, the sources and

solutions of these problems are inherently similar. Beyond any existing damage to

the target tissue, the implantation surgery introduces its own acute trauma while

micromotion of the implant, once it has been implanted, may cause chronic trauma

at the implantation site (Grill et al., 2009; Tan et al., 2013). The response window

for acute trauma in the immediate vicinity of the electrodes is on the order of 3

days (Polikov et al., 2005). During this time macrophages will phagocytose any cells

or cellular components that were sufficiently damaged during insertion. Depending

on the level of induced trauma, along with the topography and surface chemistries

of the implant, neuronal and glial cells may either grow towards or recede away

from the electrodes (Grill et al., 2009; Tan et al., 2013). Additionally, activation of

immune cells may cause rejection of the implant, leading to the formation of scar

tissue when chronically implanted (Grill et al., 2009; Polikov et al., 2005; Tan et al.,

2013). In peripheral nerve implants, if the perineurium is sufficiently damaged,

endoneurial edema, fibrosis, and nerve fiber compression may occur leading to

nerve loss in chronic implants (Bowman and Erickson, 1985; Grill and Mortimer,

2000; Grill et al., 2009; Lefurge et al., 1991). In the CNS, astrogliosis may readily lead

to implant encapsulation, causing nerve recession due to chronically released toxic

compounds from frustrated phagocytosis, which occurs when phagocytes, being

Ryan P. Sullivan 21
unable to phagocytose an object due to its size, merge together to form a

multinucleated phagocyte (Biran et al., 2007; Polikov et al., 2005; Schwartz et al.,

2006). The encapsulation layers in both the CNS and PNS inhibit both stimulation

and recording as they are insulating in nature and increase the distance between

electrode and target neurons, effectively decreasing the Signal-to-Noise ratio (SNR)

(Anderson, 2001; Grill and Mortimer, 2000).

Part of the chronic response to implants is due to continued trauma from implant

micromotion. The degree of trauma from micromotion is based on the mechanical

mismatch between the implant and its surrounding tissue (Biran et al., 2007;

Polikov et al., 2005; Suner et al., 2005); the larger the difference in stiffness, i.e.

Young’s modulus, and the sharper the implant, the more the tissue will suffer

trauma as the harder implant moves. The solution then is to make the implants as

flexible as possible, matching the target tissue’s Young’s modulus, thereby reducing

trauma (Rodríguez et al., 2000). There is a limit however, as the implant must be

sufficiently stiff and sharp enough for the actual insertion. Overly dull implants can

cause more trauma than sharp implants, while overly soft implants may not actually

penetrate to their intended depth. Techniques to overcome this include: 1) a

passive impact layer, which causes the implant as a whole to be hard enough for

implantation while keeping the outside of the implant soft, reducing micromotion

induced trauma; 2) an active impact layer, which can be activated to keep the

implant stiff during implantation but gets deactivated once implanted; and 3) an

implantation sheath that is used during surgery, which gets retracted once the

Ryan P. Sullivan 22
implant is inserted. This last option however severely limits the design possibilities

of more complex electrode arrays.

The topography, chemistries, and bioactivity of the implant have large effects on

the growth of neurons and glial cells (Tan et al., 2013). These topographical effects

include the general roughness of the material as well as localized topographies on

the nano- and micron scales. Grooves, recessions, and nanofibers on the implant

have been shown to play a role as nerve guides (Bellamkonda, 2006; Butterwick et

al., 2009; Djilas et al., 2011; Kim et al., 2008b). Control of the size and spacing of

these features have also been shown to have impeded the adhesion and

proliferation of fibroblasts (Aliuos et al., 2013; Reich et al., 2008). The chemistries of

an implant surface include small functional groups such as hydroxyl and methyl

groups, which affect the surface energy, and global and localized hydrophilicities.

Lastly, bioactivity refers to the use of larger organic molecules, usually extracellular

matrix (ECM) molecules, such as the tripeptide Arginylglycylaspartic acid (RGD) or

poly-d-lysine to target particular cellular ligands for more specific implant-cell

interactions (Huang et al., 2012; Tang et al., 2009).

Lastly, non-bound molecules of an implant also play a role as they are released after

implantation. The inclusion of Neural Growth Factors (NGF) has been shown to

promote axonal growth (Grill et al., 2009; Tan et al., 2013).

1.3.2 General Electrode Design

While each implantation site requires its own unique 3D design, they all have the

same general approach, particularly at the level of individual electrodes. The shared

Ryan P. Sullivan 23
challenges mentioned in the previous section also have shared design solutions.

Figure 1-11 depicts a cross-section of a theoretical implant electrode site. An

implant will minimally consist of a conductive component encapsulated by an

insulating bulk material. The conductive component may optionally be two

materials, a lead, which carries the electronic waveform, and an electroactive

surface, which transforms between electronic and ionic waveforms. Depending on

the fabrication method and materials used for insulating and conductive

components, an adhesion layer may be used to decrease chance of lamination for

chronic use. In the case of soft insulating substrates, an impact layer may be added

to achieve the required stiffness for implantation. Both the electroactive surface

and the surface of the insulating bulk may be modified topologically, chemically, or

biologically to facilitate desired cell interactions upon implantation as described in

the previous section. Lastly, coatings of hydrogels with chemical gradients or cells

may be used to further facilitate desired growth of the tissue immediately around

the implant (Tan et al., 2013).

Figure 1-11: Schematic cross-section of theoretical neural implant. It minimally consists of an

insulating substrate and conducting layer, which may double as the electroactive surface. Impact
layers may be added to facilitate insertion if the substrate is overly soft. An adhesion layer is used if
there are issues with adhesion between the conducting layer and the insulating substrate. Coatings
for electroactive surfaces have been of interest for their ability to drastically increase the charge
injection capacity.

Ryan P. Sullivan 24
1.3.3 Insulating materials

The insulating substrate of chronic implants needs to be flexible enough to reduce

or outright remove trauma caused by micromotion, and be hermetically sealed so

as not to allow ingress of biological fluids which may corrode or short the electrode

leads, or allow introduction of embedded bacteria during implantation (Gilletti and

Muthuswamy, 2006; Polikov et al., 2005; Szarowski et al., 2003). While silicon is

often the material of choice for in vitro electrode arrays and animal studies due its

cost effectiveness, despite the high level of development in various fabrication

techniques utilizing silicon thanks to its use in computer electronics industries, and

its ability to withstand relatively high temperatures and etching agents, it is a poor

choice in the chronic clinical setting due to its stiffness (Figure 1-12).

Figure 1-12: Logarithmic Scale Graph of Elastic Modulus of various implant materials and target
tissues. Reprinted from Scholten and Meng; Materials for microfabricated implantable devices: a
review; 2015; Lab on a Chip; with permission from the Royal Society of Chemistry. Copyright © 2015,
Royal Society of Chemistry.

The three common flexible materials used in chronic implants are

polydimethylsioloxane (PDMS; Figure 1-13) (Cheung, 2007; Delivopoulos et al.,

Ryan P. Sullivan 25
2012; FitzGerald et al., 2012; Graudejus et al., 2009; Lacour et al., 2010; Meacham

et al., 2008), poly(chloro-p-xylyene) (Parylene C; Figure 1-13) (Kim et al., 2013; Kuo

et al., 2013; Minev et al., 2015; Pang et al., 2005; Suzuki et al., 2003; Takeuchi et al.,

2005; Yu et al., 2014; Ziegler et al., 2006), and Polyimide (Figure 1-13) (A. Fomani

and Mansour, 2011; Boppart et al., 1992; González and Rodríguez, 1997; Im et al.,

2007; Lee et al., 2004; Owens et al., 1995; Peckerar et al., 1991; Rousche et al.,

2001; Shamma-Donoghue et al., 1982; Stieglitz and Meyer, 1999; Stieglitz et al.,

1997, 2000; Takeuchi et al., 2004; Zariffa et al., 2011). Of the three, PDMS is the

only one found to be used in commercially approved prosthetic devices including

cochlear implants and the Argus II retinal implant for use in a chronic clinical setting

(Tan et al., 2013). While both Parylene C and Polyimide are often used in research

settings, the lower stiffness of PDMS (0.360-0.870 MPa) compared to Parylene C

(2760 MPa) and Polyimide (2300-8500 MPa) is of significant importance for chronic

implants as seen in Figure 1-12 (Scholten and Meng, 2015).

Figure 1-13: Chemical structures of PDMS, Parylene C and Polyimide.

1.3.4 Impact Resistance/Structural Layer

Ryan P. Sullivan 26
While flexible substrates reduce trauma from micromotion, overly soft implants

introduce challenges during the implantation surgery. Overly blunt edges of an

implant will cause excessive tearing instead of cleanly slicing tissue through which it

is meant to penetrate, increasing the kill-zone, i.e. the conical section around an

electrode in which all included neurons suffer trauma to the point that they die

(Edell et al., 1992). Additionally overly soft implants may not reach the intended

penetration depths due to buckling under resistance of the target tissue (Jensen et

al., 2006). The conductive layer may naturally increase the overall stiffness of the

implant due to the nature of the materials used, primarily metals. However in cases

where this is not sufficient, a separate impact layer or tool may be introduced to

the implant for the sole purpose of increasing the stiffness (Felix et al., 2012, 2013;

Scholten and Meng, 2015). Impact layers may be passive elements, such as metal,

silicon, or ceramic, or active actuators, such as a conducting polymer or hydrogel (A.

Fomani and Mansour, 2011; Chen et al., 2009; González and Rodríguez, 1997; Kim

et al., 2014; Lee et al., 2004; Pang et al., 2005; Scholten and Meng, 2015)

(Stieghorst et al., 2014). Active actuators are particularly interesting as they not

only allow for the variable resistances needed for, and after implantation, but may

also allow implant components to alter their structure as needed over the course of

the implantation. This has been of particular interest for cochlear implants due to

the curved nature of the cochlea (Wu, 2006).

1.3.5 Adhesion Layer

Ryan P. Sullivan 27
Adhesion and seamless interfacing between the conductive layer and the insulating

substrate is of the utmost importance for chronic implants. Devices will undergo

continuous micromotion during implantation; if there are any contacts, or a gap in

contacts, this continued micromotion may cause these points of failure to spread.

Intrusion of bacteria into points before implantation may introduce infectious

agents. Intrusion by biological fluids after implantation may short the leads in the

device, or cause leakage of components that are not meant to be biological facing,

further complicating the immune response.

1.3.6 Conductive Layer and Electroactive Surface

The conductive layer and electroactive surfaces are, as with the insulating

substrate, vital to neural implants as they carry all waveforms for stimulation and

recording. The two most important aspects of conductors are their conductivity and

electrochemical stability. Materials with lower conductivities increase the signal to

noise ratio (SNR), making the choice for the conductive layer and the electroactive

surface important. The conductive layer deals purely with electric current and

therefore the electrical conductivity is important. The electroactive surface

however interfaces with biological fluid and tissue, meaning that the

electrochemical properties such as charge injection density and voltage also

become important. In many implants, including the current commercially approved

implants, the same material is used for the conductive layer and electroactive

surface (Zeng et al., 2015). This means that there is a tradeoff between electrical

Ryan P. Sullivan 28
conductivity and electrochemical stability. For example, while gold is more

conductive than platinum, and is often used in in vitro experimentation, the

increased stability of platinum makes it a much better candidate for long term

implants (Brummer et al., 1983; Cogan, 2008; Poole-Warren et al., 2010). There is

also the option of having two separate materials each specialized either for

electrical conductivity, or for electrochemical conductivity and injection capacity.

For example, recent work has shown that conducting polymer coatings on metal

conductive layers vastly increase the electrochemical conductivity of the implant as

a whole (Green et al., 2008; Guimard et al., 2007; Poole-Warren et al., 2010; Sasso

et al., 2010).

There are three general groups of electroactive materials used in implants: metal,

carbonaceous, and conducting polymer. Metals used in implants often include

platinum, titanium nitride, titanium oxide, iridium, and iridium oxide. Carbonaceous

materials include graphene, graphene oxide, carbon nanotubes (CNT), and

conductive diamond (Garrett et al., 2016). Common conductive polymers include

poly(3,4-ethylenedioxythiophene) PEDOT and poly-pyrrole (PPy). Metals have the

longest history of use in implants due to their stability, but have relatively low safe

charge injection, leading to increasing interest in carbonaceous and conductive

polymer materials; the 0.35 mC/cm2 charge injection of platinum pales in

comparison to the 242.1 mC/cm2 of graphene (Deng et al., 2011; Zhou et al., 2013).

Similarly PPy and PEDOT have been shown to decrease SNR of implants in rodent

tests (Cui et al., 2001; Ludwig et al., 2006). However, implants need to be stable on

the order of decades to be used in a clinical setting. While carbonaceous and

Ryan P. Sullivan 29
conductive polymers are promising materials to use as electroactive surfaces of

stimulating and recording electrodes to increase SNR, their long-term stability in

biological environments is yet to be proven. Many cell and rodent studies have

shown that these materials may promote neuronal survival and growth, but they

are not yet at the point of being approved for commercial use in humans (Abidian

et al., 2010; Green et al., 2009, 2012; Liu et al., 2011; Luo et al., 2011). For these

reasons, platinum and its alloys, particularly platinum:irridium (90:10), remain the

primary conductive materials used in commercial implants due to their excellent,

proven biostability at chronic scales.

While the charge injection capacity of a material may not be changed, the total

resistance of an electrode may be changed by altering its surface area. Larger

electrodes reduce the resistance, but this comes at the expense of specificity.

Smaller geometric areas allow isolation of single-unit activity from more distant

targets (Chen et al., 2009; Ludwig et al., 2006). Many commercial devices have

electrodes of considerable size, on the scale of 50-100 μm, due to the need to

penetrate tissue in order to reach their target sites. The surface area can also be

increased by roughening the surface instead of increasing the geometric area.

Platinum Gray and Platinum Black for instance are platinum surfaces that have very

different morphologies compared to smooth platinum, vastly increasing their

surface areas (Figure 1-14) (Zhou et al., 2013). This change in surface morphology

allows Platinum Gray, developed by Second Sight for the Argus II implant, to have

an injection capacity of 1 mC/cm2 compared to smooth platinum’s 0.35 mC/cm2

(Zhou et al., 2013).

Ryan P. Sullivan 30
Figure 1-14: (L-R): Smooth platinum, Platinum Gray, and Platinum Black. Scale: 10 μm. Reprinted
with permission from: Zhou, D.D., Dorn, J.D., Greenberg, R.J.; The Argus® II retinal prosthesis system:
An overview; 2013 IEEE International Conference on Multimedia and Expo Workshops, © 2013 IEEE.

1.3.7 Biologics

Biologics are bioactive components that can be used to elucidate targeted reactions

with cells. The most common set of biologics are ECM molecules (Huang et al.,

2012; Tang et al., 2009). Coating the surface of an implant with biologicals can be

used to not only encourage neurite outgrowth and inhibit the foreign body

response, but to actually segregate different neuronal populations (Lotfi et al.,

2011). For instance, in peripheral nerve implants used for arms and legs, recent

work has shown that nerve bundles may be coaxed to split into efferent and

afferent neuronal populations, thereby allowing increased specificity of stimulation

and recording. By directing neurites not only to the implant in general but to

electrode sites specifically, high signal specificity may be achieved, and may

increase the long term stability of the interface.

The other possible component for neural implants is the hydrogel and cell layer.

While biologics are confined to the surface of implants, hydrogels and other

Ryan P. Sullivan 31
coatings may be used to influence neuronal growth at distances up to millimeters

(Lu et al., 2009; Rao et al., 2012; Zhou et al., 2012). Coatings may contain gradients

of NGF and neurotropic factors to control the growth of tissue towards particular

sites on the implant before it is able to interface with any biologics. Additionally,

the hydrogel layer may be seeded with healthy cells that may integrate with the

damaged tissue to produce more favorable interfacing outcomes (Aregueta-Robles

et al., 2014; Quigley et al., 2013; Tansey et al., 2011).

1.4 DEVICE SAFETY

When considering a final device, there are four broad categories of safety that need

to be considered: 1) materials, including biocompatibility and toxicity; 2)

sterilization techniques able to be used on the device; 3) mechanical parameters

and how they influence surgical trauma and chronic micro-motion; 4)energy

exposure limits in terms of both stimulation and thermal output (Zeng et al., 2008,

Guo 2016).

For a material to be biocompatible it should be insensible or indistinguishable to

the target tissue (Guo 2016) and not produce toxins or injure tissue. Insensibility is

the ability of a material to evade detection by the host, particularly as it relates to

the immune response and scar formation. Indistinguishability is the ability of a

material to be recognized as “like” by the target tissue. Often the fastest approach

is to use materials that the FDA or other regulating body has recognized as

biocompatible. In such cases a declaration of conformity and be submitted for use

of the material. In terms of implants, some of these materials are titanium,

Ryan P. Sullivan 32
platinum, iridium, zirconium, gold, ceramic, glass, silicone rubber (PDMS),

paraylene, and Teflon. It should be noted however that materials that are

biocompatible in one instance may not be compatible in another use, such as

titanium service as an encapsulation material, but not as an electrode due to

dissolution (Guidance for Industry and FDA Staff).

An ever present risk of surgical implant of any device is the risk of infection. To

minimize this, implants should be fabrication in sterilized environments and have

sufficient integrity to survive being subjected to further sterilization methods

(Mendes et al., 2007, ISO 11135, ISO 10933-7). In the case of multipart devices, it is

also important to ensure that there are no gaps between materials in which a

bacterial biofilm can form. In 2002, there was a recall of cochlear implants after

patients presented with bacterial meningitis, attributed in part to such biofilm

formation (Wei et al., 2007).

The size, configuration, and mechanical properties of implants are important

parameters that affect trauma both during and post implantation. In addition to the

importance of the elastic modulus matching the target tissue as shown in Figure 1-

12 (Gong et al., 2016) the implant must have the proper geometry to appose the

target tissue. Failure to ensure this apposition may cause additional trauma, such as

with the mention meningitis cases where such trauma provide a direct path for the

meningitis to propagate (Wei et al., 2007). While this may be minimized by having

rounded soft edges of insulating coating, the total density and tethering of the

implant is also an important consideration (Lind et al., 2012). The elastic absorption

Ryan P. Sullivan 33
of the coating may not be sufficient for high accelerations the patient may

experience, so it is important to both minimize the mass and anchor or secure the

implant if possible, such as to the skull or in the case of the cochlear implant, in the

bone bed behind the pina (Guo 2016).

In terms of energy limits, both the electrochemical and thermal outputs need to be

considered. The thermal dissipation is highly dependent on the local

vascularization, but in call cases, implants should never reach 39 0 C in vivo (Guo

2016). Electrochemical limits are more complicated. Two key limits were

determined in early testing of platinum electrodes. The first is the electrochemical

limit known as the water window described by Rose and Robble (Rose et al., 1990).

It is so named as it is the range outside of which water hydrolysis occurs, producing

H2 in the negative phase and O2 in the positive phase. In vitro, the water window

may cover 50-100 μC/cm2/phase, though in vivo ranges may differ by a factor of 10

due to protein absorption onto the electrode surface (Robblee et al., 1980, Cogan

et al., 2016). , This occurs due to the absorption inhibiting damage opposed to

actually altering the window (Hibbert et al., 2001)

The second is the tissue damage threshold described by Shannon [2] (McReery et

al., 1990):

log(𝐷) = 𝑘 − log⁡(𝑄) (1-1)

where D is the charge density and Q is the charge per phase. The value of k

therefore describes a boundary between damaging and non-damaging stimulation

regimes based on empirical data from a number of studies, with a value of 1.85 for

Ryan P. Sullivan 34
k is a strong qualitative boundary (Cogan et al., 2016). However there are severe

limitations of the Shannon equation. In particular it is based only on platinum

macroelectrodes and does not explicitly account for duty cycle, pulse frequency,

current density, non-uniform currents, alternate electrode materials, and

microelectrodes.

Current density thresholds have been shown to vary with pulse width and

frequency (Butterwick et al., 2007). Densities as low as 12μC/cm2 have been shown

to be damaging, while densities of 60μC/cm2 have been found to not produce

apparent histological damage (McCreery et al., 1990, MeCreery et al., 2006),

disagreeing with the 30μC/cm2 from the water window and Shannon threshold. It

has been found that there is a micro-macro boundary between 200 and 300 μm

diameter (3x10-4 and 7x10-4 cm2). While the Shannon equation roughly holds true

for electrodes above this size, many current neural implants have considerably

smaller diameters so does not apply (Cogan et al., 2016). At this smaller size the

charge per phase becomes the critical with 4nC/phase being the qualitative

threshold (McCreery et al 1994, McCreery et al 2010). It is important to note

however that this still depends on pulse frequency and duty cycle, and further

research is needed to elucidate the interplay of all of the waveform

parameters(Cogan et al., 2016).

1.5 FABRICATION TECHNIQUES

Each of the materials and components described in Sections 1.3.1-1.3.8 have a

variety of fabrication methods best suited to them individually. The challenge

Ryan P. Sullivan 35
comes in fabricating an implant that is able to have many or all of these

components at once. The focus for the next sections of this literature review will be

on those materials that are currently commercially available, or appear that they

may become commercially available in the foreseeable future. For flexible

insulating substrates, PDMS is the most appropriate target. For the conductive

components, the metals platinum and platinum:iridium (90:10) alloy will be the

focus. Conductive polymers, which may serve both as active actuators for the

impact layer as well as electrode coatings for the electroactive layer will also be

covered.

1.5.1 Deposition Overview

The use of flexible polymers as substrates is severely prohibitive for many

traditional metal deposition techniques. Many of the procedures and techniques

are borrowed from the semiconductor industries, which focuses on silicon

(Scholten and Meng, 2015). Unlike silicon, PDMS has a low degradation

temperature of 350°C and is incompatible with many masking and etching

combinations. While platinum and iridium are used for their long term stability in

the final implant, they also provide additional challenges requiring higher

temperatures, 1772°C and 2447°C respectively, for thermal deposition and stronger

etching agents (Atkins, 1989; Moulton et al., 2012). For these reasons, low

temperature additive fabrication methods are more beneficial as they do not

require the use of masks and etching agents.

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In terms of fabrication, neural implants capable of stimulation and recording may

broadly be categorized into two types: planar 2D electrodes, and more complicated

3D electrodes. Planar electrodes are used for retinal implants, optic nerve implants,

and PNS cuff electrodes (Yue et al., 2016). It is important to note that the planar

classification refers particularly to the fabrication of the device; the optic nerve and

PNS cuff electrode implants are fabricated in a single plane, but are subsequently

bent for the actual implantation. Planar electrodes also include shank electrodes,

often used for cortical experiments; however these only target a small subset of

neurons (Yue et al., 2016). More complex 3D electrodes are needed when the

targeted neuronal populations are spread out over a 3D space, such as in cochlear

implants, afferent-efferent PNS implants, and cortical implants. The fabrication

requirements are different for each category of implant, with planar fabrication

seeing the most development.

Soft polymer fabrication often includes micromachining or using 3D molds. For

planar electrodes, like the retinal implant, fabrication techniques include spin

coating or chemical vapor deposition (CVD) of the polymer layer, followed by

deposition and photolithography of the metal layer, followed again by another

polymer layer to seal the leads (Delivopoulos et al., 2012; FitzGerald et al., 2012;

Graudejus et al., 2009; Lacour et al., 2010; Meacham et al., 2008). The latter

polymer layer is then etched, often with O2 reactive ion etching (RIE), to expose the

end of the leads and electroactive surfaces; alternatively photopatternable PDMS

can also be used (Delivopoulos et al., 2012; Lacour et al., 2010). In the case of

PDMS, the time required to etch and expose the electroactive surfaces can be

Ryan P. Sullivan 37
prohibitively long, as laser ablation offers poor resolution and control (Scholten and

Meng, 2015), and the melting temperature of the metals used precludes thermal

evaporation. Similarly an alternative to deposition techniques requiring high

temperatures incompatible with polymer substrates are micromachined platinum

foils. PDMS substrates are cast first cast, then laminated with a thin foil of the metal

of choice before being covered by a second layer of PDMS. A pulsed laser is then

used to ablate the PDMS substrate (Ordonez et al., 2015, Green et al., 2014).

Additionally, gas permeability may introduce bubbles during fabrication depending

on the morphology of the conducting layer, which may result in delamination or

shorting of the electrodes (Scholten and Meng, 2015). Currently, for 3D structures

such as cochlear or cortical implants, molds are often used to set the polymer

material around conductive components. Molds may still often require etching to

expose the leads, and need to be custom made for each individual design. This

becomes problematic as there is large variation on a patient to patient level in

general anatomy even before the differences in trauma are accounted for that

caused the need for the prosthetic. Custom fabrication of neural implants designed

specifically for each patient may increase stimulation and recording fidelity by

allowing closer apposition between the electrode and target tissue.Additive

fabrication techniques are a promising avenue for overcoming the limitations

imposed by fabricating PDMS and platinum in the same device. By nature, additive

fabrication deposits materials locally, not requiring subsequent masks or etching

steps. Additionally, recent work in additive fabrication has demonstrated the ability

to print not just the insulating substrate and metal conductors, but also the ability

Ryan P. Sullivan 38
to deposit a range of conducting polymers, carbonaceous materials, and various

biologics including ECM molecules and cell populations (O’Connell et al., 2015;

Wagner et al., 2013). Two such additive fabrication techniques are Dip-Pen

Nanolithography (DPN) and Inkjet printing. DPN covers scales of 5 nm - 5 μm, and

has been shown to print ECM molecules (Figure 1-15) (Curran et al., 2010;

O’Connell et al., 2015). Inkjet covers scales of 1 μm - 100 μm, and has been shown

to be able to print insulating substrates, metals, and cells (Figure 1-15) (Ferris et al.,

2013a, 2013b; Perelaer et al., 2009; Reinhold et al., 2009; Stieghorst et al., 2016;

Yamazaki et al., 2007).

Figure 1-15: Scale of DPN and Inkjet fabrication capabilities based on volume of droplets deposited.
The number in parenthesis is the diameter of a drop at the given volume to give an estimate of the
resolution. The diameter will naturally change with contact angle based on the ink and substrate
used.

1.5.2 Dip-Pen Nanolithography

Dip-Pen Nanolithography (DPN) is a direct-write lithographic technique based on

Atomic Force Microscopy (AFM) that uses AFM cantilevers to deposit inks to a

substrate in a manner not unlike the old writing quill. The pyramidal heads of the

cantilevers are dipped into ink wells to cover the cantilever in ink. When the tip is

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brought sufficiently close to the substrate, a meniscus forms and transfers the ink

as shown in Figure 1-16.

Figure 1-16: DPN ink transfer. 1) Tip between features; 2) contact with substrates causes meniscus
to form. Diameter of features is proportional to the square root of dwell time. 3) Rupture of capillary
bridge determines the volume of a sphere during retraction (O’Connell et al., 2014); 4) Deposited
feature takes upon the form of a spherical cap after reaching equilibrium.

For biological molecules, DPN can be used to print features at densities 10,000 to

100,000 higher than other printing methods (Ginger et al., 2004). Certain cellular

phenomena are quite sensitive down to scales of 50-100 nm; it is therefore

important to be able to decorate a surface with sufficient resolution to properly

arrange biomolecules when spatial positioning is so vital for eliciting desired

function, such as in the case of RGD motifs (Curran et al., 2010). With suitable ink

formulations, metal and polymers may also be printed (Nakashima et al., 2012),

opening up possibilities for all-in-one additive fabrication of neural electrodes.

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There are two major types of DPN ink, molecular and liquid. In the case of

molecular inks, the cantilever is repeatedly inked, allowing the ink to dry between

applications, forming a layer of deposited ink. At a sufficiently high humidity, a thin

layer of water forms on the substrate and cantilever (Ginger et al., 2004). This

water causes some of the deposited ink to go into solution, which can then be

transferred through the meniscus formed when the tip approaches the substrate

(Rozhok et al., 2003).

The other type of ink is liquid ink, for which special cantilevers have been designed

to contain ink wells. While molecular inks make use of the water from high

environmental humidity to act as the ink carrier, liquid inks contain their own

carrier. When the tip of the cantilever is dipped in an ink well, the ink migrates up

the tip, and along the underside of the cantilever through the specially designed

channel into the ink reservoir (NanoInk, Inc., 2012). The tip may then be placed in

contact with a given substrate, where the ink will flow from the reservoir back

down the tip due to capillary action (O’Connell et al., 2014).

Due to the scale of DPN, the technique lends itself more to patterning the surface

of existing electrode substrates rather than to the fabrication of entire devices.

Work with PDMS inks has demonstrated the ability to control the topography on

the substrate by depositing pillars down to 2.7 μm in diameter and 18.6 nm in

height (Hernandez-Santana et al., 2011). While there has been much research into

depositing conductors using DPN, both metal such as platinum and gold, and

polymer such as PEDOT:PSS, DPN work has mostly been limited to the printing of

Ryan P. Sullivan 41
organized grids instead of full conductive lines (O’Connell et al., 2015). This is due to

the fact that DPN ink transfer is an inherently passive transfer, relying purely on the

fundamental forces such as the surface energies of the substrate, ink, and pen to

drive the transfer of the ink, making continuous homogenous lines difficult to

deposit on the hydrophobic surfaces used as insulating substrates.

Electrodeposition is precluded due to the insulating nature of the flexible

substrates, and due to etching reagents for platinum and iridium being too

damaging for the polymer substrates (O’Connell et al., 2015). The biggest and most

successful application of DPN has been the complex patterning of biomolecules

(Senesi et al., 2009). DPN offers an unparalleled ability to pattern biomolecules such

as DNA, proteins, or ECM molecules with nanometer resolution (Kim et al., 2008a;

Loh et al., 2008). While research has been done on cell printing with DPN, so far it

seems to be limited to single bacterium due to the small size of the DPN tips (Kim et

al., 2012).

1.5.3 Inkjet

Inkjet printing is an additive fabrication technique that functions by creating

pressure waves in an ink chamber, resulting in formation and expulsion of ink

droplets toward a substrate (Wijshoff, 2010, 2012). This process can be continuous,

where droplets form at a given frequency with no pause, or Drop-on-Demand

(DOD), where droplet formation can be arbitrarily controlled, though still often at a

set frequency. Drop-on-demand is the natural choice for additive fabrication, as it

Ryan P. Sullivan 42
allows for more complex, arbitrary patterns to be printed (Wijshoff, 2012). The

method by which droplets are formed and expelled fall into 4 general categories:

thermal, piezoelectric, electrostatic, and acoustic (Wijshoff, 2012). The piezoelectric

approach has received much of the research focus due to not having to worry about

thermal or charge limits of the inks used (Wijshoff, 2012). As shown in Figure 1-17,

piezoelectric inkjets function by expanding and contracting a piezoelectric material

connected to a main ink chamber; in this particular case, the piezo contracts,

expanding the chamber drawing in ink, before subsequently expanding, causing the

ink chamber to expel ink, causing droplet formation and expulsion (Figure 1-17).

Figure 1-17: Schematic of the interior of single piezoelectric inkjet chamber.

Piezoelectric inkjet heads were traditionally limited in their resolution to 1pL ink

chambers, which roughly corresponds to a 10 μm diameter of the resulting droplets

on a substrate. Recent work, in what is called Super Inkjet Technology, has pushed

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the size of inkjet chambers down to 0.1 fL, corresponding to 500 nm diameter of

deposited droplets, and rivaling the upper scales of DPN (Murata et al., 2005;

Sekitani et al., 2008). Inkjet printer cartridges often consist of an array of individual

jetting chambers, with each chamber being able to fire at frequencies between 1-20

kHz depending on the particular design of chamber and ink formulation used

(Wijshoff, 2012).

Inkjets have been shown to be able to print a variety of materials, including flexible

polymers, metals, conductive polymers, and even living cells (Xiong and Liu, 2012).

PDMS ink formulations are usually either thermally cured or cured using ultraviolet

(UV) light. While thermally cured formulations are approved for medical implants,

the curing process is time consuming, though recent work with long-wave infrared

lasers for heat transfer have reduced this time down to 2 s (Chen et al., 2006;

Efimenko et al., 2002; Stieghorst et al., 2016). Furthermore, laser irradiation of

PDMS does not seem to affect its biocompatibility (Green et al., 2010). Much work

has been done with metal nanoparticle inks, particularly silver (Fribourg-Blanc et

al., 2013; Layani et al., 2012; Reinhold et al., 2009). Nanoparticles are capped with

short organic molecules to prevent aggregation, which may be removed with

sintering temperatures well below the melting point of the bulk metals (Reinhold et

al., 2009). Less reactive metals, such as platinum and iridium, have not however

enjoyed much printability, as even in nanoparticle form, the sintering temperatures

are still prohibitively high for soft flexible substrates (Reinhold et al., 2009). The

mechanics and design of inkjetting will be discussed in further detail in Chapters 3,

4, and 5 (also see Sun et al., 2016; Wijshoff, 2010). An in depth review of biological

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inks, and conductive polymer inks is out of the scope of this thesis, however further

details on both of these can be found in Gudapati et. al. (2016), and Weng et. al.

(2010) respectively.

1.5.4 Metal Inks and the Polyol Process

There are three types of metal inks used in both DPN and Inkjet printing: metal

nanoparticle, metal ligand, and metal salt inks. As seen in Figure 1-18, nanoparticle

inks consist of solid metal particles coated with capping agents needed to stabilize

the particles in solution and prevent aggregation. Metal ligand inks consist of metal

atoms bound to organic ligands while salt inks consist of dissolved metal salts

(Figure 1-18), often chlorides. Common metals used include copper, silver, and gold

(Chang et al., 2012; Dearden et al., 2005; Huang et al., 2003; Lee et al., 2007; Leff et

al., 1995; Liu et al., 2005; Magdassi et al., 2003, 2010; Valeton et al., 2009).

Nanoparticle inks are typically capable of metal loadings of 20-70%, while metal

ligand and metal salts are typically around 30% metal loading (Wünscher et al.,

2012, 2014). Nanoparticles are however more prone to oxidation and aggregation,

limiting the lifetime of inks, as well as resolutions on the submicron scale due to

clogging, or particles being larger than the resolution achievable with DPN

(Wünscher et al., 2014). Salt inks are completely homogenous by nature, and can

therefore be scaled down without issue.

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Figure 1-18: Schematic representation of the three types of metal inks. Since this thesis work has
focused on platinum, the schematic representation is of the 3 different types of platinum inks that
are possible.

Once deposited, all three inks must be sintered to form conductive tracks. In the

case of nanoparticle inks, the capping agents must be removed from the particles,

usually with thermal excitation, allowing the particles to coalesce. Ligand inks

similarly require removal of the ligand, which will reduce the metal, allowing

coalescence (Wünscher et al., 2014). Metal salts require a relatively high

temperature compared to the other two methods, in order to thermally decompose

the salt, thereby evaporating off the anions. While all three of these techniques

have produced conductive traces requiring temperatures lower than the melting

point of bulk metal, they are still limited to only being printed on more resistant

flexible materials such as Polyimide (Wünscher et al., 2014). Silver is the most

commonly used metal for printing due to its high conductivity and low

temperatures. However, even silver inks can require temperatures over 200°C to

reduce nanoparticle inks (Deng et al., 2013; Greer and Street, 2007; Lee et al.,

2005), 125-200°C for silver metal-ligand inks (Dearden et al., 2005; Perelaer et al.,

2009; Smith et al., 2006), and 300-500°C for thermal decomposition of salt inks such

Ryan P. Sullivan 46
as AgNO3 (Liu et al., 2005; Mei et al., 2005). The metals used in commercial

implants, platinum and iridium, require even higher temperatures, having melting

points of 1768°C and 2447°C respectively, which significantly limits the

development of inks for these metals.

Recent work on alternative sintering methods has been developed to replace or

reduce the thermal requirements for sintering of deposited inks. These alternative

approaches include chemical, plasma, electrical, and photonic sintering methods

(Wünscher et al., 2014). Chemical sintering methods involve the application of a

second solution which facilitates the sintering. In the case of nanoparticle inks, the

added chemicals usually serve to destabilize or replace the capping agents, freeing

the nanoparticles to agglomerate and sinter (Wakuda et al., 2007, 2009). A

technique called reactive inkjet-printing may also be used to deposit a second ink

containing a reducing agent that can fully reduce and sinter metal-ligand and metal

salt inks (Chen et al., 2012; Kao et al., 2011, 2012; Kheawhom and Foithong, 2013;

Smith and Morrin, 2012; Valeton et al., 2009). Alternatively the substrate may be

modified with reducing agents in a technique known as click chemistry; this

technique is often used for DPN, but as it may alter the entire substrate, it goes

against the localized approach of additive fabrication (Jung et al., 2004). Plasma

sintering (Figure 1-19) is a low temperature approach which reduces metal-ligand

and metal-salt inks directly when using reducing or inert atmospheres, and removes

nanoparticle capping agents due to the low pressure of the vacuum and high

surface energy caused by the plasma, thus increasing the rate of surface diffusion

(Bromberg et al., 2013; Crowther and Badyal, 1998, 2012; Reinhold et al., 2009).

Ryan P. Sullivan 47
Electrical sintering may also reduce metals directly; however it requires a

conducting substrate and is therefore not conducive to insulating polymer

substrates. The most promising approach is photonic sintering, which seeks to

stimulate the metal ink while minimally interacting with the substrate as a whole;

however two different approaches are currently being used for photonic sintering.

The first approach involves global stimulation, in which lamps such as infrared (IR)

and UV irradiation are used to reduce the metal inks, as they are more strongly

absorbed by the metal inks than their transparent substrates (Cherrington et al.,

2011; Denneulin et al., 2011; Fritsch et al., 2013; Jahn et al., 2010a, 2010b; Tobjörk

et al., 2012). The second approach involves the use of targeted lasers, allowing for

focused local sintering of an ink. In an ideal case, the stimulation wavelength should

be matched to the highest absorption peak of a given ink (Bieri et al., 2003; Cheng

et al., 2009; Ko et al., 2007b, 2007a; Maekawa et al., 2012; Watanabe and

Miyashita, 2007; Yeo et al., 2012; Yun et al., 2008). This is particularly useful as the

sintering approach may then use the same device as the laser annealing of PMDS

mentioned in the previous section. For an in depth review on these alternative

sintering methods, see Wünscher et al. (2014).

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Figure 1-19: Mechanism of plasma reduction and sintering of salt inks.

During the early stages of the work for this thesis, we developed a new type of

metal salt ink called a polyol ink (O’Connell et al., 2014). This is a chemical approach

whereby the ink solvent is itself a reducing agent, in this case belonging to the

polyol family. While the chemical sintering methods mentioned above usually seek

to reduce the ink immediately after it is deposited, the reducing polyol solvent does

not significantly react with the metal salt at ambient temperature. Functionally this

means that stable inks can be created which require significantly lower sintering

temperatures to reduce the ink to a conductive metal, compared to regular metal

salt inks, but stays stable during printing. Bulk platinum melts at 1768°C, while it’s

metal salt ink H2PtCl6 thermally decomposes and sinters at 500°C (Schweizer and

Kerr, 1978); by using a polyol ink with Ethylene Glycol as the polyol, this can be

reduced further to just 160°C (Cho and Ouyang, 2011), which is considerably closer

to the limits of flexible substrates such as PDMS. Polyol inks may be further

combined with the previously mentioned approaches, such as plasma and photonic

Ryan P. Sullivan 49
sintering, thus making up the remaining difference in the thermal envelope

(O’Connell et al., 2014).

To our knowledge, our previous work was the first documented use of a polyol ink

for fabrication; however the polyol process in itself has been extensively studied,

including the use of platinum and iridium (Anumol et al., 2011; Bonet et al., 1999;

Fanavoll, 2013; Harish et al., 2012; Hei et al., 2012; Li et al., 2005; Patel et al., 2005,

2006; Shobin and Manivannan, 2014; Sinha and Sharma, 2005; Song et al., 2007),

and is one of the major methods for the synthesis of nanoparticle inks. In the

literature, a wide range of metal salts have been found to work with the polyol

process as seen in Figure 1-20 and Table 1-1. During synthesis particles will

naturally aggregate and coalesce, so capping agents such as polyvinylpyrrolidone

(PVP) are used to maintain a narrow distribution of particle sizes. As the desired

result for fabrication is aggregation, the parameters from nanoparticle synthesis

studies that do not produce tight distributions may in fact be the desired ones for

creating solid continuous films.

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Figure 1-20: Table of elements with metals labeled by viability of use in polyol process. Elements in
green have been demonstrated to create reduced films. Elements in yellow have been shown to
reduce to nanoparticles, though no work has been done to create films. Elements in blue reduce to
metal oxide are therefore not viable without further steps.

Overall, the polyol process is a promising approach for deposition of both single

metals such as platinum, and combined alloys, such as platinum:iridium on soft

flexible substrates (Joseyphus et al., 2007b, 2007a). Continued research into the

effects of pH, salt and polyol concentration, and thermal gradient may allow tight

control over nucleation and growth of metal films (Bonet et al., 1999; Cho and

Ouyang, 2011). When combined with expansions of inkjet such as Super Inkjet

Technology and focused photonic reducing and sintering methods, polyol inks may

allow for large scale deposition of drops with 500 nm resolution, paving the way for

rapid fabrication of full neural implants.

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Table 1-1: Survey of metal salt and reducing agent combinations appearing in literature.

Metal Metal Salt Reducing Agent Reduction Method Study Details Morphology Reference
Particle (20 nm)
Nickel Ni(OAc)2 EG or TEG Thermal 120-200°C, 1 hr (Kurihara et al., 1995)
Film (9-15nm)
Cu2O Glycerol Thermal 220°C, 60 min Particle (1 μm) (Sinha and Sharma, 2002)
Cu(OH)2 Glycerol Thermal 220°C, 60 min Particle (3.5 μm) (Sinha and Sharma, 2002)
Copper Particle (10-80 nm)
EG or TEG Thermal 120-200°C, 2 hr (Kurihara et al., 1995)
Cu(OAc)2 Film (12-42 nm)
Glycerol Thermal 234°C, 60 min Particle (5.6 μm) (Sinha and Sharma, 2002)
Particle (12.1 nm)
CoCl2 EG or TEG Thermal 120-200°C, 2 hr (Kurihara et al., 1995)
Film (15-23 nm)
Cobalt
Co(OAc)2 EG or TEG Thermal 120-200°C, 2 hr Particle (14 nm) (Kurihara et al., 1995)
Co(C11H23COO)2 1,2-butanediol Thermal 175°C, 20 min Particle (>200 nm) (Atmane et al., 2014)
EG or TEG Thermal 120-200°C, 1 hr Particle (5 nm) (Kurihara et al., 1995)
150°C Particle (Bonet et al., 1999)
Thermal 160°C 90 min, 180°C 15 min,
Particle (Teranishi and Miyake, 1998)
EG 198°C 15 min
RuCl3
900 W, 2450 Hz Particle (Fanavoll, 2013)
Ruthenium Microwave
30 s to color change Particle (Teranishi and Miyake, 1998)
DEG Thermal 245°C, 5 min to color change Particle (Teranishi and Miyake, 1998)
TREG Thermal 285°C, 2 min to color change Particle (Teranishi and Miyake, 1998)
EG Thermal 180°C Particle (2.1 nm) (Joo et al., 2010)
Ru(acac)3
1,4-butanediol Thermal 175°C Particle (2.8-3.1 nm) (Joo et al., 2010)
Na3RhCl6 EG Thermal 140°C Particle (Zettsu et al., 2006)
140°C Particle (Watson et al., 2015)
185°C Particle (Zhang et al., 2010)
Thermal
EG 190°C, N2 atmosphere Particle (Hei et al., 2012)
Rhodium
RhCl3 120-200°C, 1 hr Film (9 nm) (Kurihara et al., 1995)
Thermal NT: 70-80°C Particle (Biacchi and Schaak, 2011)
DEG Thermal NT: 85-95°C Particle (Biacchi and Schaak, 2011)
TREG Thermal NT: 95-105°C Particle (Biacchi and Schaak, 2011)

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 Metal Metal Salt Reducing Agent Reduction Method Study Details Morphology Reference
TEG Thermal NT: 100-110°C Particle (Biacchi and Schaak, 2011)
Thermal NT: 80-90°C Particle (Biacchi and Schaak, 2011)
EG
Thermal 185°C Particle (Zhang et al., 2010)
RhBr3 DEG Thermal NT: 105-110°C Particle (Biacchi and Schaak, 2011)
TREG Thermal NT: 110-120°C Particle (Biacchi and Schaak, 2011)
TEG Thermal NT: 115-125°C Particle (Biacchi and Schaak, 2011)
EG Thermal NT: 105-110°C Particle (Biacchi and Schaak, 2011)
DEG Thermal NT: 120-130°C Particle (Biacchi and Schaak, 2011)
Rh2(TFA)4
TREG Thermal NT: 130-140°C Particle (Biacchi and Schaak, 2011)
TEG Thermal NT: 135-145°C Particle (Biacchi and Schaak, 2011)
[Rh(Ac)2]2 EG Thermal 185°C Particle (Zhang et al., 2010)
Rh(acac)3 EG Thermal 185°C Particle (Zhang et al., 2010)
EG Thermal 190°C, N2 atmosphere Particle (Hei et al., 2012)
Methanol Particle (Teranishi and Miyake, 1998)
H2PdCl4
Ethanol Particle (Teranishi and Miyake, 1998)
1-propanol Particle (Teranishi and Miyake, 1998)
EG Microwave 800 W 2450 MHz Particle (Patel et al., 2005)
Na2PdCl4
Glycerol Microwave 800 W 2450 MHz Particle (Patel et al., 2005)
Particle (10 nm)
Palladium EG or TEG Thermal 120-200°C, 1 hr (Kurihara et al., 1995)
Film (18-22 nm)
EG(10%)/Glycerol
PdCl2 Microwave 800 W, 2 min Particle (Li et al., 2013)
(90%)
EG(10%)/Glycerol
Microwave 800 W, 2 min Particle (Li et al., 2013)
(90%)
Pd(NO3)2 EG Thermal Room Temperature Particle (Bonet et al., 1999)
Pd(NH3)4(NO3)2 EG Thermal 100°C Particle (Bonet et al., 1999)
Thermal 160°C Particle (Sun et al., 2001)
EG Thermal 160°C Particle (Renoirt et al., 2014)
Silver AgNO3 Microwave 800 W, 2450 MHz Particle (Patel et al., 2005)
Particle (40 nm)
EG or TEG Thermal 120-200°C, 1 hr (Kurihara et al., 1995)
Film (34-50 nm)

Ryan P. Sullivan 53
 Metal Metal Salt Reducing Agent Reduction Method Study Details Morphology Reference
175°C Particles (1.5-2.5 μm) (Sinha and Sharma, 2005)
Thermal (Shobin and Manivannan,
Glycerol 200°C Particle
2014)
Microwave 800 W, 2450 MHz Particle (Patel et al., 2005)
SnCl2 EG Sonication Particle (Qiu, 2005)
Tin
SnO EG or TEG Thermal 120-200°C, 1 hr Particle (36 nm) (Kurihara et al., 1995)
Particle (8 nm)
WO3 EG or TEG Thermal 120-200°C, 3 hr (Kurihara et al., 1995)
Film (12 nm)
Tungsten
Particle (10 nm)
Na2WO4 EG or TEG Thermal 120-200°C, 3 hr (Kurihara et al., 1995)
Film (12 nm)
Rhenium Re3Cl9 EG or TEG Thermal 120-200°C, 1 hr Film (14 nm) (Kurihara et al., 1995)
Thermal 100°C Particle (Bonet et al., 1999)
Iridium H2IrCl6 EG
Microwave 900 W, 2450 Hz Particle (Fanavoll, 2013)
Plasma N2 Metallic Film (O’Connell et al., 2013)
110°C Black Particle (Bonet et al., 1999)
Particle (2 nm)
120°C, 2 hr (Khelashvili et al., 2006)
Film (10-15 nm)
150°C Particle (Bonet et al., 1999)
160°C, 15 min Particle (Cho and Ouyang, 2011)
EG 170°C Particle (Joseyphus et al., 2010)
Thermal
120°C, 5 min Dark Film
H2PtCl6
140°C, 5 min Metallic Film
Platinum
160°C, 5 min Metallic Film (160 nm) (Sun et al., 2010)
180°C, 5 min Metallic Film
200°C, 5 min Metallic Film
190°C, N2 atmosphere Particle (Hei et al., 2012)
190°C, 20 min (NH4)2CO3 to
EG (vapor) Thermal Porous Film (Gopalan et al., 2015)
make porous dry H2PtCl6
EG or TEG Thermal 120-200°C, 1 hr Particle (Kurihara et al., 1995)
EG Microwave 800 W 2450 MHz Particle (Patel et al., 2005)
Na2PtCl4
Glycerol Microwave 800 W 2450 MHz Particle (Patel et al., 2005)

Ryan P. Sullivan 54
 Metal Metal Salt Reducing Agent Reduction Method Study Details Morphology Reference
PtCl2 EG Thermal 160°C Particle (Sun et al., 2010)
Thermal 120°C Particle (Bonet et al., 1999)
EG
Plasma 30 W, 1100 mTorr, N2 Film (O’Connell et al., 2013)
(Shobin and Manivannan,
Glycerol Thermal 200°C Particle
HAuCl4 2014)
Gold but-2-yne-1,4-diol Particle (Jeyabharathi et al., 2014)
but-2-ene-1,4-diol Particle (Jeyabharathi et al., 2014)
butane-1,4-diol Particle (Jeyabharathi et al., 2014)
Particle (28 nm)
AuCl3 EG or TEG Thermal 120-200°C, 2 hr (Kurihara et al., 1995)
Film (32 nm)
100°C, 170°C, 2 min Particle (10-150 nm) (Joseyphus et al., 2010)
EG Thermal
120°C Particle (100 nm) (Joseyphus et al., 2007a)
FeCl2 PG Thermal 120°C Particle (Joseyphus et al., 2007a)
Iron
Particle (60 nm)
TMEG Thermal 120°C (Joseyphus et al., 2007a)
Fe3O4 formation
Fe(OAc)2 EG or TEG Thermal 120-200°C, 2 hr Particle (20 nm) (Kurihara et al., 1995)

Abbreviations: EG: Ethylene Glycol; DEG: Diethylene glycol; TREG: Triethlyene Glycol; TEG: Tetraethlyene Glycol; PG: Propylene glycol; TMEG: Trimethylene glycol

Table 1-2: Plasma reduction of metal salts.

Metal Metal Salt Plasma Atmosphere Study Details Morphology Reference

25 μm penetration
H2/He 100 W, 5 min, generation of HCL gas (Koo et al., 2006)
Palladium PdCl2 depth
H2 (Shim et al., 2007)
10 W, H2O solvent water electrode, generation Black Particles at
H2/He (Koo et al., 2005)
of HCL interface
Platinum H2PtCl6
He 10 W, H2O solvent water electrode no reduction (Koo et al., 2005)
Ar Particles (Wang et al., 2009)

Ryan P. Sullivan 55
1.6 RATIONALE

The overall goal of this thesis was to develop a platinum deposition method that

could be used as an adjunct method to the existing additive fabrication methods for

silicone substrates, conducting polymer covers, and printing of biologics and cells.

By focusing only on materials that are already approved for commercial

implantation, particularly platinum and silicone, the additive fabrication method

developed in this thesis may be more rapidly brought to use in clinical settings. The

goal then will be to create a metal deposition method that is scalable from the

nanometer to 100 μm scales, with deposition rates that are scalable and sufficiently

rapid for use in industry. It must also be amenable to other additive fabrication inks

meaning it must work at ambient temperature and pressure with localized

deposition and treatment that can be controlled in an ad-hoc manner allowing for

tailoring to the needs of individual clinicians and patients. Furthermore additive

fabrication techniques will lower the cost of medical implants due to lower material

wastage and labor costs. By building upon our previous work with DPN and polyol

inks (O’Connell et al., 2013, 2014), low temperature deposition and reduction of

polyol ink may in principle not only allow for the deposition of platinum on silicone,

but allow arbitrary alloys, such as platinum:irridum to be also printed on silicone,

which are of interest as the combined allow is has better long-term stability than

platinum alone.

1.7 AIMS AND OBJECTIVES

1.7.1 General Aim

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The general aim of this study was to build on our previous work with platinum

polyol inks, and extend it to printing on soft flexible PDMS substrates. Furthermore

the aim was to be able to print not only grids of features, but to fabricate lines of

platinum at high speeds.

1.7.2 Specific Aims

The specific aims of this research were:

(i) To determine the difference in the parameters required to switch from

DPN printing of platinum on silicon, to printing of platinum on PDMS, in

addition to printing lines on PDMS.

(ii) To develop an analytical method to ensure the high accuracy of inkjet

printing at scales that would be required for industrial fabrication.

(iii) To determine the viability of extending polyol inks beyond DPN printing

to Inkjet printing for industrial upscaling, by developing inkjet

waveforms that can reproducibly print 10 and 1 pL droplets of platinum

ink at high speeds with low error.

(iv) To maximize the speed and resolution of printing stable lines with inkjet

printing to approach micron-scale resolution.

1.8 SIGNIFICANCE

While exact numbers are difficult to come by, there are currently approximately 3.3

million people with significant visual impairment in the United States that may

warrant visual implants (Congdon et al., 2004). Additionally there are also an

Ryan P. Sullivan 57
estimated 185,000 amputations (1998), 12,000 spinal cord injuries, and 1.9 million

cases of Tramatic Brain Injury (TBI) per year in the US alone, that may warrant

peripheral nervous system implants, spinal cord implants, and cortical implants.

Cochlear Ltd. implanted 26,838 implants last year with expectations of continued

growth, with the company’s total revenue estimated to pass 1 billion this year for

Cochlear Ltd alone (Cochlear Ltd., 2016). Massive interest in the growing field of

prosthetics has led to the US Food and Drug Administration to recently release

technical considerations for additive manufactured devices for new fast tracked

approval. The prosthetic device industry is already a billion dollar industry with

cochlear implants alone, and will continue to grow to multibillions as new implant

types become commercially approved.

Ryan P. Sullivan 58
CHAPTER TWO: DIP-PEN
NANOLITHOGRAPHY OF PLATINUM INK
ONTO POLYDIMETHYLSILOXANE

2.1 INTRODUCTION

Cell interactions fundamentally take place at the nanoscale. While cell bodies may

be on the scale of 30 μm in diameter, components like axons or dendritic spines

have diameters around 500 nm and 50 nm respectively, and it is important to be

able to control materials at these scales (Curran et al., 2010; Loew and Hell, 2013;

O’Connell et al., 2015). Nanoscale fabrication techniques are therefore vital for

both basic research and clinical applications for resolving and controlling these

nanoscale interactions. Dip-Pen Nanolithography (DPN), an Atomic Force

Microscopy (AFM) technique that directly deposits materials with cantilever tips, is

particularly suited for controlling surface topography and chemistry at the

nanoscale. It has been shown to be able to deposit biomolecules such as, DNA, RGD

motifs, and extracellular matrix (ECM) molecules down to the 10 nm resolution

(Curran et al., 2010; Ginger et al., 2004; Loh et al., 2008). DPN has also been used to

modify the topography of silicone at single micron widths and ~20 nm in height

(Hernandez-Santana et al., 2011), deposit conductive polymers (Wagner et al.,

2013), and even deposit metals such as platinum (O’Connell et al., 2014). However,

due to its passive transport mechanism, fabricating lines has proven to be quite

Ryan P. Sullivan 59
difficult, and often requires click chemistry (Long et al., 2007; O’Connell et al.,

2014).

In our previous work, we analyzed the fluid dynamics of ink between cantilever

wells and tips that was responsible for the sizes of deposited features (O’Connell et

al., 2014). We additionally demonstrated the ability to print nanoscale platinum

patterns on silicon substrates using a polyol ink (O’Connell et al., 2013). These two

studies provide the foundation for the work done in this chapter. Our previous

work focused on the effect of dwell time on feature size of a platinum polyol ink,

over a range of 0.002 to 5 s, after 10 min N2 plasma reduction at 1100 mTorr,

resulting in features with diameters between 50 nm and 2 μm. This chapter will

show that the approach and withdraw phases of DPN deposition are also important

factors in determining the final feature size, and are vital when depositing on

hydrophobic substrates like PDMS.

2.2 EXPERIMENTAL DETAILS

2.2.1 PDMS Substrate Creation

Clean glass slides were etched using a diamond tipped pen. They were then

sectioned by applying force to pieces along the length of the etched lines,

producing clean cuts. Once the desired number of pieces were obtained, they were

then rinsed in filtered water and sonicated for 10 min. The glass slides were

subsequently dried under a N2 flow.

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The following is a time-sensitive protocol and was performed exactly in the order

described. Figure 2-1 is a graphical representation of the following steps.

Glass petri dishes 3 cm in diameter were cleaned, and immersed in water with

Palmolive Original Dishwashing Liquid (Colgate-Palmolive, USA) for 1 hour to allow

a surfactant film to build on the glass. This was necessary to allow easy removal of

final PDMS product. The water was drained from each petri dish and the tip of a

Kimwipe (Kimtech Science*, USA) was used to absorb the small amount of

remaining water in the dishes and rupture any bubbles that had formed. The dishes

were then air-dried for a further 2 hr (Figure 2-1(1)).

1 g MED-4234 Part A (NuSil, USA) was added to a glass vial with screw top lid, and

dissolved in 10ml n-hexane by alternating between 30 s of vortexing and 30s

manual stirring with a metal spatula until completely dissolved. The solution was

subsequently sonicated for 30 s to remove any dissolved gases.

Glass pieces were rinsed with water and sonicated for 10 min (Figure 2-1(2)). They

were subsequently rinsed with IPA and sonicated for 10 min (Figure 2-1(3)) and

then dried with N2. Cleaned glass pieces were then placed in a Plasma Cleaner

(PDC-002, Harrick, USA) and were cleaned using atmospheric plasma at 30 W for 10

min, allowing for the creation of additional hydroxyl groups on the glass to which

the PDMS can bind (Figure 2-1(4)). The glass pieces were then placed in the dry

petri dishes in a fume hood in preparation for the PDMS solution to be poured

(Figure 2-1(5 and 6)).

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0.1 g of MED-4234 Part B was then added to the MED-4234 Part A/n-hexane

solution. The solution was manually swirled for 10 s to ensure it was thoroughly

mixed. The resulting solution was then poured into the prepared petri dishes in the

fume hood to a depth of 1 cm, completely covering the prepared glass slides (Figure

1-2(7)). The PDMS solution was left in the fume hood overnight to allow the solvent

to evaporate (Figure 1-2(8)).

The following morning the petri dishes containing the glass slides, now coated with

a thin film of uncured PDMS (Figure 1-2(8)), were placed in a thermal oven at 120°C

for 3 hr (Figure 1-2(9)). The petri dishes were removed, and left to cool to room

temperature. Once cooled, a scalpel blade was used to cut the now cured PDMS

film along the edges of the glass pieces to allow the removal of the glass-PDMS

substrates (Figure 1-2(10)). Each substrate was marked at one end, cleaned under a

stream of distilled water for 10 s, dried with N2 (Figure 2-1(11)), and then placed in

individual cleaned 1.5 mL Eppendorf tubes for storage until required (Figure 1-

2(12)).

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Figure 2-1: Graphical representation of the steps required to prepare PDMS samples for subsequent
fabrication.

Ryan P. Sullivan 63
2.2.2 Polydopamine Synthesis

PDMS samples were plasma treated (PDC-002, Harrick, USA) for 10 min using

atmospheric plasma at 30 W and 1100 mTorr to add hydroxyl groups onto which

the dopamine could react and anchor.. The PDMS samples were then placed in a

solution of dopamine in Tris Buffer pH 8.5 (1 mg/mL) and were gently agitated on

an orbital shaker for durations ranging from 10 min to 24 hr. The PDMS samples

were subsequently removed, rinsed under deionized water for 10 s, and dried

under N2 flow.

2.2.3 Ink Formulation

It was discovered early on in the production of the platinum ink for this study that

the H2PtCl6 used to make the ink was highly corrosive, as it corroded the metal

spoons being used to scoop the H2PtCl6 out of its container. We therefore

improvised and cut the tops off plastic Pasteur pipettes to create makeshift scoops.

These scoops were used to place 0.794 g of H2PtCl6 (Sigma-Aldrich) in 1.5 mL

Eppendorf tubes; 0.7 g of ethylene glycol was subsequently added to the Eppendorf

tubes resulting in a 25% metal loading. It should be noted that this metal loading

refers specifically to the Pt by weight, not the entire H2PtCl6 salt. This is equivalent

to the loadings utilized in our previous study (O’Connell et al., 2013), however our

previous study reported the metal loadings in terms of the total salt. The reason for

expressing the metal loading specifically in reference to the Pt by weight was so

that it was directly comparable to metal ink loadings used in other studies, such as

those using nanoparticle inks. As mentioned above, the reactive nature of the metal

Ryan P. Sullivan 64
salts causes them to react with metal spatulas, negating their use. It was also found

that the inks would react within the months after preparation, gradually turning a

darker brown color as opposed to the bright orange of fresh ink as seen in Figure 2-

3.

Figure 2-2: Aging inks were found to be a problem if kept over periods of months. The left picture is
of bulk ink creation. The right picture has two inks; the left ink was made 10min before the image
was taken while the right ink was made 6 months before the image was taken.

2.2.4 DPN Modification

DPN cantilevers (NanoInk PEN-0300-03, material Si3N4, length 107 μm, width 22

μm, spring constant 0.5 N/m; Figure 2-3) were cleaned by soaking in IPA for 10 min,

then rinsing in IPA, before being dried under N2 stream. The cantilevers were then

used as is, or were further modified with a hydrophobic octadecyltrimethoxilane

(ODTMS) coating (O’Connell et al., 2014). The cantilevers that were modified with

Ryan P. Sullivan 65
ODTMS were treated for 10 min using atmospheric plasma at 1100 mTorr at 30 W

with a Plasma Cleaner (PDC-002, Harrick, USA), before being immersed in 0.1%

ODTMS in IPA for either 5 min or 1 hr. The cantilevers were subsequently left to dry

for 30 s before being placed in an oven heated to 80°C for 5 min. This process was

repeated with the cantilevers being immersed and heated for 5 min each, 3 more

times. This modification was simultaneously performed on silicon wafers to allow

for a visual verification of hydrophobic modification via changes in water contact

angle to be conducted.

Figure 2-3: Picture of 12 DPN tips on a cantilever. The distance between tips, 66 μm, was an
important factor in printing. No pattern was created wider than 50 μm so as to remove possible
interactions between the adjacent tip and already deposited features.

Ryan P. Sullivan 66
2.2.5 DPN Fabrication

DPN ink wells (NanoInk Universal Inkwell IWL-0009-03) were cleaned in IPA with 10

min sonication, rinsed with IPA, and then were dried by N2. 1.5 μL of the prepared

platinum salt ink was then pipetted into the ink reservoirs on the DPN ink wells.

Using an AFM (MFP-3D Classic, Asylum Research, USA), DPN cantilevers (NanoInk

PEN-0300-03) were mounted and leveled on the main pad of the ink well chip. This

was done by manually lowering the DPN array until the tips became visibly

deflected. The chip was then adjusted until the right most tip, i.e. the tip that

appeared in the lower part of the monitor, deflects first (Figure 2-4). This was done

so that deviations in the polymer substrate, in this case PDMS, would not adversely

affect printing by interacting with non-printing DPN tips. Once leveled, the laser

was aligned to the bottommost pen, which was then calibrated on silicon substrate

before use.

Figure 2-4: Optical images of a DPN array using the internal camera of an Aslyum AFM. The left
image shows the cantilever leveled so that no tips touch while the right image shows the cantilever
leveled so that when the tip closest to the bottom of the screen touches the substrate, the other tips
are less deflected.

Ryan P. Sullivan 67
The bottommost tip was then manually lowered into an inking station, where it was

lowered until minimally visibly deflected. After 10 s, the cantilever was quickly

raised, and the frequency was measured to detect the level of ink loading. The

substrates used included silicon wafer, PDMS, and PDMS-PDA. The cantilever was

moved over the substrate, and manually lowered down to the substrate to make a

large bleed spot. This bleed spot was used as a landmark and point of origin (0,0)

for deposited grids so that they could be easily found with later analytical methods

such as optical profilometry, AFM, and scanning electron microscopy (SEM).

In the case of grids, the cantilever was then moved left across the substrate in 50

μm increments, and a force grid of 4 x 5 points was taken at each increment. The

Aslyum AFM does not have a built in system for liquid ink DPN printing, so contact

force scans had to be taken in a manner similar to DPN. By controlling the applied

force and dwell time of a series of force curves, DPN like control could be

established. Forces of 7, 10, and 15 nN were used with dwell times of 0.5, 1, 2, 8,

10, 20 s.

In the case of lines, a contact image was performed with the cantilever locked to

the same row. In this way, the line material may be deposited with each pass.

2.2.6 DPN Feature Characterization

Deposited grids were immediately reduced with N2 plasma for 10 min before being

measured with Veeco optical profilometry and Asylum AFM (MFP-3D Classic,

Asylum Research, USA). The bleed spot was found and used as the main landmark

for finding the deposited grids. Optical Profilometry was done using a x100

Ryan P. Sullivan 68
magnification in VSI mode in a grid pattern starting at the bleed spot and moving

left. In this way, even if grids were not readily apparent with the normal optics,

topographical data of the region where the grids should have been deposited was

still collected. AFM scans were taken in contact mode in air using a similar grid

pattern for the same reason.

As will be discussed in the analysis section, there were no detected grids deposited

under a range of parameters. To further rule out just missing the features due to

their size, optical profilometry and AFM was also done in the same manner on

deposited grids immediately after deposition, forgoing the reduction step. The N 2

plasma reduction naturally reduces the volume of the deposited features, so the

larger unreduced features should be easier to detect if they were in fact deposited.

2.3 RESULTS AND DISCUSSION

2.3.1 PDMS Casting

Glass samples which did not undergo 10 min atmospheric plasma treatment before

PDMS casting frequently underwent delamination during handling. Plasma

treatment was therefore used to stabilize the PDMS and remove any air pockets.

The introduction of detergent was used to stop the PDMS from bonding irreversibly

with the glass petri dishes. It was found that adding too much detergent beyond

that needed to coat the surface of the petri dishes would negatively affect the

PDMS. As seen in Figure 2-5A, samples became cloudy when excess detergent was

used. By using capillary action of Kimwipes (Kimtech Science*, USA) to remove the

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last remaining liquid from the petri dishes, this problem was rectified, producing

clear PDMS liquid (Figure 2-5B).

Figure 2-5: Pictures of two petri dishes filled with PDMS precursors. (A) PDMS precursor solution is
cloudy white, due to extra surfactants on the glass surface going into solution. (B) PDMS precursor
solution is clear as required.

PDMS samples were cured at temperatures of 80°C, 100°C, and 120°C for 3 hours. It

has been shown that the silicon hydride in PDMS precursor curing agents is able to

reduce metal salts such as AgNO3, HAuCl4, and H2PtCl6 (Goyal et al., 2009; Simpson

et al., 2003; Zhang et al., 2008). This reaction can be enhanced by using a second

curing session once the PDMS has interacted with the metal salt, thereby enhancing

the reduction as well as completing the curing of PDMS (Dunklin et al., 2013, 2014).

It was thought that this reductive mechanism might increase the adhesion of the

metal salt ink to the substrate and result in stronger bonding to the PDMS. It was

found that the adhesion between the substrate and the DPN tip was greatly

increased. This allowed the PDMS to be deformed, causing it to stay attached to the

DPN tip during retraction. This deformation resulted in stretched peaks roughly

Ryan P. Sullivan 70
200nm tall as seen in Figure 2-6. 120°C was chosen as no deformation of the

substrate was observed when printing.

Figure 2-6: AFM image of a partially cured PDMS substrate that has adhered to the DPN tip, causing
large deformations as the PDMS was raised when withdrawing the tips from the substrate.

2.3.2 PDMS Surface Modification with Dopamine

PDMS substrates were modified with PDA films using a solution of 1 mg/mL

dopamine HCl in Tris Buffer with pH 8.5. This was done to both increase the

hydrophilicity of the substrate, and to facilitate a form of click chemistry. The

catechol groups in PDA have been shown to be able to directly reduce metal salts.

This reductive ability was thought to be able to enhance the deposition and final

adhesion of the Pt. Polymerization times ranged from 10 min to 24 hr. Longer

polymerization durations were found to result in deposited PDA particles which

adhered to the DPN tip and impeded printing. 10 min was the optimal time chosen

as it resulted in no PDA particles forming on the PDMS.

PDMS surfaces were characterized by optical microscopy before and after PDA

modification as seen in Table 2-1.

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 Table 2-1: Surface roughness values on unmodified and PDA modified (1 hr) PDMS substrates. The top two rows are at x50 magnification while the bottom two
rows are at x100 magnification. See Section 2.5 Appendix for a full explanation of how the values were calculated.

2
Magnification Sample Rt Ra Rms Skew Kurtosis Variation(μm ) Entropy Entropy Deficit
PDMS 28.34 ± 10.33 1.74 ± 0.53 2.39 ± 0.73 0.39 ± 1.18 5.50 ± 5.33 16.55 ± 0.87 -18.57 ± 0.28 0.10 ± 0.07
x50
PDA 47.99 ± 20.32 3.12 ± 5.39 3.11 ± 1.22 1.37 ± 0.89 7.60 ± 7.95 28.35 ± 6.26 -18.34 ± 0.32 0.12 ± 0.06
PDMS 13.00 ± 4.68 1.43 ± 1.76 1.78 ± 1.75 0.51 ± 0.68 2.28 ± 2.80 6.97 ± 3.49 -19.03 ± 0.27 0.05 ± 0.06
x100
PDA 32.61 ± 17.27 1.85 ± 1.06 2.28 ± 0.56 1.92 ± 0.68 9.69 ± 4.81 9.90 ± 1.24 -18.67 ± 0.24 0.17 ± 0.06

Ryan P. Sullivan 72
2.3.3 DPN Deposition

2.3.3.1 NanoInscriptor to Asylum

It is important to differentiate the two instruments used during the candidature.

The original DPN used was a NanoInk NScriptor. This was a device designed

specifically for DPN and was used in our previous work (O’Connell et al., 2013)

during the early parts of the candidature. It however broke at the same time that

the NanoInk went bankrupt and was not able to be repaired.

Subsequent work, as described in this chapter, was done using an Asylum AFM.

Unlike the NScriptor, the Asylum was not a dedicated DPN device, lacking much of

the automated controls and special functions that would facilitate DPN

experiments.

The substrates, inks, and DPN tips used were identical for both devices. Initial

deposition experiments on the Asylum were performed using cleaned but

unmodified DPN cantilevers on PDMS cured at 120°C for 2 hr, with 25% platinum

loading ink. Beyond the initial bleed spot, deposited grids were not readily

detectable when measured either before or after plasma treatment. We explored

the same parameter ranges as previously performed on the NScriptor in an attempt

to produce deposition. No change in dwell times, 0.1-10 s, or applied forces, 7-15

nN, produced any differences in the results.

2.3.3.2 Effect of Tip Modification

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It was previously found that ink may migrate off the DPN tip onto the main chip of

the cantilever when the cleaning steps included plasma cleaning (O’Connell et al.,

2013), resulting in no deposition. As each tip was only used for a single printing

session and was not reused, plasma treatment of the DPN tips was not a part of the

protocol used in the present study. However, ODTMS modification of the

cantilevers was still performed in an attempt to get printing working. The exact

methods that we had used previously (O’Connell et al., 2013) were replicated, with

an additional set of DPN tips also being modified for the longer duration of 1 hr.

Additionally optical profilometry was performed on DPN tips immediately after

inking, before any deposition occurred, to confirm the presence of ink at the tip end

as seen below in Figure 2-7.

Figure 2-7: Optical images of (A) DPN tip array and the edge of the chip; and (B) zoomed in image of
the left most chip in panel A. These images show that the metal ink does not run off the tip unlike
previously seen in our past work (O’Connell et al., 2013). We attribute this to the fact that we had
previously used plasma cleaning in order to reuse the tips. In this work, all tips were single use only.
Both unmodified and ODTMS modified DPN tips showed that ink remained on the tips with no
evidence for any on-chip migration over all of the arrays surveyed.

While ODTMS modification of the test silicon wafers was shown to be successful via

visual confirmation of changes in water contact, neither set of ODTMS modified

Ryan P. Sullivan 74
DPN tips resulted in successful deposition of the ink on PDMS. Every optical image

taken showed that the DPN tip reservoir was full and that no ink migrated onto the

larger chip (Figure 2-7). As there was no ink migration on the unmodified tips, the

purpose of the ODTMS modification was rendered moot and was no longer used.

DPN deposition was also attempted on clean silicon substrates, although it too was

shown to be unsuccessful. Neither AFM nor SEM found any deposited features on

the clean silicon substrates beyond the initial bleed.

2.3.3.3 Effect of Withdrawal Speed

Around the time this work was being undertaken, Chad Mirkin’s group at

Northwestern University (the pioneers of DPN) released a paper on the effect of

hydrophobicity of the substrate during the approach and withdrawal phases

(Eichelsdoerfer et al., 2014). Most of the previous research in the literature,

including our own, had focused on the ink transfer during the dwell phase, where

the DPN tip is in static contact with the surface of a substrate (O’Connell et al.,

2013). However Chad Mirkin’s group showed that not only the time in which the ink

on the DPN tip is in contact with the surface, but the time during which the tip is

approaching or withdrawing from the substrate must also be taken into account

(Eichelsdoerfer et al., 2014). In cases of hydrophilic surfaces with polar solvents, the

integral of the ink transfer over these two time periods is additive; however, in the

case of hydrophobic surfaces with polar solvents, as is the case here, the integral of

ink transfer over these time periods is actually subtractive (Eichelsdoerfer et al.,

2014). That is to say, as the tip is retracting from the hydrophobic surface, the ink

Ryan P. Sullivan 75
will migrate back up onto the cantilever. In principle then, on hydrophobic

substrates, sufficiently slow withdrawal speeds may allow sufficient time for the

entirety of the ink to recede back onto the tip and thereby not produce deposited

features.

It is unknown what the withdrawal speed of the DPN tips was in our previous

NanoInscriptor DPN which was used to develop the platinum approach (O’Connell

et al., 2014), however the default speed of the Asylum AFM profile was 1 μm/s. An

applied force of 10 nN, dwell times of 2 s, and approach and withdrawal speeds of 1

μm/s, 10 μm/s, and 100 μm/s were tested. At 1 μm/s no deposition was achieved

(Figure 2-8A, D). At 10 μm/s however, controlled deposition was achieved (Figure 2-

8B, E) with features having average widths of 2.26 ± 0.30 μm and heights of 17.47 ±

7.73 nm. In the case of 100 μm/s, the DPN tip actually indented the soft PDMS

substrates (Figure 2-8C, F). We attribute this to requiring longer times before the tip

is slowed down sufficiently for printing at higher speeds. Features deposited at 100

μm/s had average widths of 0.91 ± 0.25 μm and depths of -7.73 ± 4.17 nm.

Deposited ink was found around the holes created by the tip (Figure 2-8C), but

were difficult to measure.

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Figure 2-8: Effect of withdrawal speed on deposition with DPN. Images A, B, and C are profiles of
single features in D, E, and F respectively. 1 μm/s approach and withdrawal speed produces no
deposition as can be seen in panels A and D. 10 μm/s approach and withdrawal speed produces
deposition as seen in panels B and E. 100 μm/s approach and withdrawal speed causes indentation
and deformation of the PDMS surface as seen in panels C and F. Scale bars 10 μm.

DPN deposition on PDA modified PDMS was able to achieve similar cross sections

with lower dwell times. At 10 μm/s approach and withdrawal, 2s dwell time, 10 nN

applied force, the average feature widths were 1.72 ± 0.13 μm with heights of

202.61 ± 25.77 nm, significantly larger than on PDMS (Figure 2-9A, D)). We attribute

this to the higher hydrophilic of the PDA, as well to its reactive chemistry. Without

using plasma treatment, it was found that the ink would react with the substrate,

forming a solid at the ink-substrate interface that was reflective under optical

profilometry. Contact mode AFM 1 hour after deposition was found to dislodge

~90% of the deposited material leaving toroidal features around 20 nm in height

(Figure 2-9 B, C, E, F). These toroidal structures survived 10 min water sonication,

while the ~90% upper portion were removed by even 10s of water stream. The 1

hour delay is incidental; it was the time between printing multiple samples,

Ryan P. Sullivan 77
performing optical profilometry, before returning to AFM to do a contact mode

scan.

Figure 2-9: AFM images taken in AC mode of grids printed on PDMS-PDA substrate. A-C are height
profiles of D-F. Panels A and D are the profile and height map of a representative grid printed with 2s
dwell 10 nN applied force. Panels B and E are the profile and height map of a representative grid
printed with 2s dwell 10 nN applied force that has been scanned with contact mode AFM. After 1 hr
of contact with the substrate, it was found that contact mode would dislodge the higher part of the
features leaving toroidal structures. Panels C and F show zoomed in scans of the remaining toroidal
structures.

2.3.3.4 Line Printing on PDMS-PDA without Plasma Reduction

Line printing was attempted on both normal PDMS and PDA modified PDMS. Scan

speeds of 0.5, 1, 5, and 50 μm/s were used with forces of either 10 or 25 nN. This

was achieved by taking a contact image and holding the tip in the same line,

continuously tracing for the desired time. A small amount of ink residue, not line

deposition was achievable on bare PDMS at any speed or applied force. This was

similar to results previously found by our group using silicon substrates (O’Connell

et al., 2013). Line deposition was achievable on PDA modified PDMS, with the line

Ryan P. Sullivan 78
width found to decrease greatly with each line printed, exhibiting a similar bleed

effect as grid printing, as seen in the optical profilometry images in Figure 2-10

below. Figure 2-10A and B are the height map and intensity images from the optical

profilometer, while C and D are height profiles of the thicker bottom line and

thinner upper line respectively. The intensity image is effectively an optical image

that one would obtain with a light microscope. These two lines, each printed at 25

nN, 5 μm/s, 90 μm length, and 20 min deposition time, produced respective

average widths of 10.76 and 4.71 μm, with peak heights of 175.6 and 111.4 nm,

demonstrating the bleed like effect where subsequent lines are smaller. Beyond the

apparent trends, it was difficult to fully quantify the deposition mechanics due to

only 2 full lines being able to be printed per inking. Tertiary lines and beyond would

leave residue, or discontinuous lines that were difficult to resolve. The ends of the

lines, where the cantilever slows down and comes to a stop before reversing

direction, were slightly larger than the mid sections. This was attributed to the

contact area being able to grow larger with the longer contact times. The optical

images showed that most of the deposited material was highly reflective, indicating

reduction of the metal ink into solid metal. However the peak points on the height

map (Figure 2-10A) have corresponding dark points on the intensity map (Figure 2-

10B) indicating that there may be unreduced material at the ends of the lines and

at certain local maxima in the larger lines. Unlike plasma treatment, catechol

facilitated reduction would take place at the PDA-ink interface, working bottom to

top. Any limit in reduction therefore would manifest on the lines once they reached

a certain thickness, effectively the penetration limit, causing further deposited

Ryan P. Sullivan 79
material to not reduce. This effect can also be seen in the two heights maps (Figure

2-11 A and B) with their intensity maps (Figure 2-11 C and D) in the double lines

Figure 2-11. The high thickness of the Figure 2-11A results in almost completely

black, i.e. unreduced layer as seen in Figure 2-11C.

Figure 2-10: DPN deposited lines on PDA modified PDMS. The top images are representative height
map (A) and intensity map (B) of deposited lines. The profiles C and D are transverse profiles of the
thicker bottom line and thinner top line respectively. As can be seen in B, the majority of the lines
are highly reflective, while tall deposits, particularly at the ends of the lines, are black. These black
regions correspond to the local peaks in the height map (A). Scale bars: 50 μm.

For unknown reasons, certain DPN tips produced bisected lines (Figure 2-11)

instead of single lines (Figure 2-11). This occured at both 10 nN and 25 nN, and at

both 5 μm/s and 50 μm/s. It appeared to be dependant on a given tip, since

multiple inkings and depositions with a given tip will result in either single or double

lines, but not both. As seen in Figure 2-11A, C, and E, shorter lines and slower rates

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produce thicker depositions. As total deposition time is held constant, the length of

a line is inversely proportional to the amount of time a given part of the line is in

contact with the liquid on the tip. In Figure 2-11 the shorter 0.5 μm/s line (A, C, E)

has a total width of 107 μm while the longer 5 μm/s line (B, D, F) has a total width

of only 5.6 μm. Lines printed at 50 μm/s (Figure 2-12), did not form continuous

depositions due to the high speed at which they were printed.

Figure 2-11: Optical Profilometry images and profiles of two lines. Images A, C, and E are of a line
printed at 0.5 μm/s while images B, D, and F are of a line printed at 5 μm/s. Panels A and B are

Ryan P. Sullivan 81
height maps; C and D are optical images; E and F are transverse height profiles. As can be seen the
base material is highly reflective (C and D), though deposited material above a certain height does
not reduce and appears black. This can be seen more clearly in the 0.5 μm/s line (panel C) where the
majority of the surface is black. It is unknown what causes certain lines to be bisected in the middle.
Scale bar: 10 μm.

2.3.3.5 Line Printing on PDMS-PDA with Plasma Reduction

While deposited material may have already been reduced by the PDA film, it still

resembled many individual particles (Figure 2-12A). Lines that underwent 10 min N2

plasma treatment after deposition showed altered morphology, and produced solid

continuous films in parts of the line where sufficient material was present (Figure 2-

12B). Plasma environments have previously been shown to sinter metal

nanoparticles by increasing the surface energy and facilitating surface diffusion

(Reinhold et al., 2009). The plasma treatment in the present study however,

seemed to negatively affect the PDA film. As seen in Figure 2-12B and C (blue

arrows), the PDA fractured under the plasma treatment. This was attributed to loss

of water molecules as a result of the vacuum of the plasma environment (Zhang et

al., 2013).

Figure 2-12: AFM contact mode height maps of 50 μm/s samples that did, and did not undergo
plasma treatment. A) Samples without plasma treatment show a morphology resembling a stack of
many individual particles. We attribute this to difference growth planes as the platinum would be
pulled out of the ink and grow from the substrate upwards; B) Samples treated with 10 min N 2
plasma coalesced into a single film where sufficient material was present (i.e. the rightmost part of

Ryan P. Sullivan 82
the line). Where there was not sufficient material, islands formed. C) Larger scale image of PDMS-
PDA substrate shows cracks. The blue arrows in B and C point to cracks that were observed in the
PDMS-PDA substrate and were attributed to the plasma treatment. The cracks are brighter, rather
than darker, because the edges fold up, similar to a dried up lake (Zhang et al., 2013). Optical
profilometry can have difficulty with sharp valleys, detecting only the edges. Scale bars: (A) 5 μm; (B)
4 μm; (C) 10 μm.

2.3.4 Scanning Electrochemical Microscopy

As the PDA is not a strong conductor like Au-Mylar used in later chapters of this

thesis, direct electrical connection was not able to be established to perform

Conductive-AFM. In order to obtain information about the conductivity then,

scanning electrochemical microscopy (SECM) was attempted on lines deposited on

PDMS-PDA substrates. It was found however that the redox mediator solution

reacted with the samples and produced a black residue over the entire substrate as

seen in Figure 2-13. This unfortunately precluded any measurements being made

on the platinum lines. Multiple samples were attempted and produced the same

result. A 10 min IPA sonication was shown to be able to remove much of the visible

residue, leaving the highly reflective deposited platinum lines intact, thus

demonstrating a strong adhesion of the platinum lines to the PDMS-PDA substrate.

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Figure 2-13: Optical images of SECM adverse sample reaction with redox mediator solution. A)
Deposition/reaction of SECM solution with PDMS-PDA substrate. B) After 10 min IPA sonication;
most of the material had been removed, but the deposited platinum remained. Red arrows point to
the line array of which measurement was attempted. Scale bar: 500μm.

2.4 CONCLUSION

The hydrophobic and soft nature of PDMS makes it a difficult substrate on which to

print polar inks such as the polyol ink used in the present study. However, by tightly

controlling the window of the applied force, and the cantilever speed, we were able

to successfully and reproducibly deposit grids of platinum. Due to the passive

nature of DPN deposition, liquid inks do not lend themselves to line printing beyond

atomic layers. By introducing a chemically reactive coating, we were able to deposit

platinum lines over 100 nm with successive passes. However the extreme

fabrication time of 20 min for a single line may make up-scaling the fabrication of

lines difficult, though lower print times may be used for thinner lines. Furthermore

difficulties in exactly predicting the line widths and heights from applied force will

need to be remedied with further research, similar to the many papers analyzing

the fluid flow used during static contact, commonly used in grid printing.

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2.5 APPENDIX

2.5.1 Calculation of Roughness Values

RMS (σ), skewness (γ1), kurtosis (γ2) are calculated from central moments of the i-

th order μi:

1⁄
2
𝜎 = 𝜇2

𝜇3
𝛾1 = 3⁄
2
𝜇2

𝜇4
𝛾2 = −3
𝜇22

Variation is the integral of the absolute value of the local gradient.

The Shannon differential entropy for a probability density function is

𝑆 = − ∫ 𝑝(𝑥) log 𝑝(𝑥) 𝑑𝑥

𝑋

𝑛
𝑝𝑖
𝑆 ≅ − ∑ 𝑝𝑖 log
𝑤𝑖
𝑖=1

Where X is the domain of the variable x.

2.5.2 Overlapping drops

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Figure 2-14: Optical profilometry height map of features deposited using 10 s, 10 nN, 10 μm/s
parameters of PDMS-PDA. During the 10 s contact time, the ink reacts with the substrate. As the tip
attempts to deposit subsequent drops, it interacts with the previously deposited drop, causing it to
merge into the existing meniscus. This has the effect of moving a droplet along in discrete steps
every 10 s or so (including the withdraw, lateral movement, and approach between each dwell). As
can be clearly seen, what then remains are overlapping discs of material that have reacted with the
PDA surface. The large feature is the remaining ink left during the final retraction.

Ryan P. Sullivan 86
 CHAPTER THREE: AUTOMATION OF
CHARACTERIZATION OF INKJET DROPLETS
USING IMAGEJ SOFTWARE

3.1 INTRODUCTION

Automated software acts as a force multiplier on research by decreasing the time

spent on data collection and analysis, increasing reproducibility by removing human

bias and fatigue, and allowing ever expanding datasets to be collected, and analysis

techniques to be performed. However much of the software used are closed source

black boxes, skipping the peer-review process, which is antithetical to science

(Morin et al., 2012; United States National Science Foundation, 2010). Closed

source software often has issues with reproducibility and external validation,

causing errors to go unnoticed for years, to the detriment of research based on

those and subsequent findings (Adams, 1984; Easterbrook and Johns, 2009; Hatton

and Roberts, 1994; Ince et al., 2012). Even in cases where errors are found, they

may remain unfixed, requiring new versions to be purchased (Microsoft, 2015).

Given how often labs delay deployment of updated equipment and software as a

cost-saving measure, these errors may persist for decades, judging by the number

of labs still running XP era software. In US preclinical trials alone, an estimated 28.2

billion USD is wasted every year due to irreproducibility, of which 7.191 billion USD

Ryan P. Sullivan 87
(25.5%) is attributed to errors and inadequacies in data analysis and reporting

(Freedman et al., 2015).

Open source software remedies these issues by virtue of being free, removing

barriers to use, and being open to peer-review, allowing external validation and

improvements. As Linus’s Law goes, “given enough eyeballs, all bugs are shallow”

(Raymond, 1999). To this effect, the US National Institutes of Health has funded

ImageJ and its predecessor Image, open source public domain image processing

programs, since 1987 (Schneider et al., 2012). ImageJ is plugin based software,

allowing researchers to easily extend and share far beyond the core functionality of

the program as needed. Distributions and use cases cover many fields including

biology, astronomy, and materials science and work with multiple characterization

techniques including Scanning Probe Microscopies (AFM, SPM, etc.), Light

Microscopies (Brightfield, Fluorescent, Confocal, etc.), Electron Microscopies (SEM,

TEM, etc.), and even Astronomical Telescopies (Schindelin et al., 2012, 2015;

Schneider et al., 2012).

Of the aforementioned microscopy techniques, light microscopy is the main

approach used to quantify drop-on-demand fabrication processes such as inkjet

printing. There are generally four approaches used to quantify Inkjet:

(i) The wettability of a given substrate by the desired ink can be measured using a

horizontal microscope to determine the contact angle. This determines the

diameter of drops, and therefore the ultimate resolution and distance required

between dots.

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(ii) Drop formation can be monitored using high speed cameras placed orthogonal

to the jetting plane. In this method a strobe light is flashed at a frequency matching

the piezo contractions so that images formed from the integration of many droplets

are captured at the desired stage of formation (Dong et al., 2006a, 2006b, 2007).

This allows the stages of droplet formation, ejection, break-away, and coalescence,

to be studied in detail. Parameters such as fluid properties, nozzle geometry, and

waveform dictate a large part of these effects (Gan et al., 2009; Kwon, 2009, 2010;

Liou et al., 2009; Shin et al., 2011; Tekin et al., 2008; Wijshoff, 2010).

(iii) Drop impact and spreading kinetics are studied in a similar manner, having a

high speed camera placed orthogonal to the plane of the substrate. In the initial

impact stage, which occurs during the first 1 μs, the kinetic energy is partially

dissipated by viscous forces. Over the next 0.1 – 1 ms, the remaining energy is

driven into the surface, causing the droplet to spread to an increased diameter.

Depending on the strength of the kinetic energy, the size of the drop may oscillate

before reaching a stable size (Cummins and Desmulliez, 2012).

(iv) Test patterns of deposited material can be characterized by top-down light

microscopy or photography to detect printing accuracy and errors. This can be done

in real-time between print layers for error detection and correction using an

integrated camera, or can be done post printing using a separate imaging device to

assess accuracy and viability of a particular jet combination. While software is

widely used within the printing community which currently has a heavy focus on

visual perception, it is not readily available for the materials community (Cummins

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and Desmulliez, 2012). The real-time case also requires special proprietary

hardware and software which can be exceedingly expensive.

We present in this Chapter, an open source software solution, extending ImageJ

that is able to access the accuracy of test patterns using light microscopy data. We

have developed a naïve algorithm not requiring a priori knowledge of dot position

to quantify grid order (or disorder) based on relative dot positions along with

morphological data. This may be combined and extended with μManager,

microscope control software that interfaces with ImageJ, to create a complete, all-

encompassing solution to the automation of data collection and analysis of inkjet

test patterns, facilitating rapid ink and waveform development (Edelstein et al.,

2010). This software will be extensively used in Chapters 4 and 5, providing an

analysis platform allowing full quantitative characterization of patterns when

combined with line quantification and contact angle plugins (Stalder et al., 2006,

Stalder et al., 2010, Bickford 2013).

3.2 EXPERIMENTAL DETAILS

A platinum polyol ink (H2PtCl6/ethylene glycol 10% Pt by weight) was created as

previously described in in Chapter 2, and as will be produced for use in the work of

Chapters 4 and 5. A 10 pL Diamatix Inkjet Printer cartridge was filled with 1 mL of

the ink and used to print calibration grids of 6x5 dots. The spacing between dots

was 80 μm with an expected dot size of around 20 μm. These grids were then

imaged with a x25 magnification objective using a Veeco Optical Profilometer.

Ryan P. Sullivan 90
We developed an ImageJ plugin using the Fiji Macro builder and Notepad++ to then

analyze these grids. Grids resulting from both stable and unstable jetting regimes,

where the nth droplet is either approximately identical to the 0th droplet or not

respectively, along with grids on both pristine and noisy substrates, that is either

clean homogenous substrates or heterogeneous substrates with imperfections,

debris from sources such as ink splatter, were analyzed to test the bounds of the

software. These were used to experimentally validate parameters and methods for

automatic background removal, thresholding, object detection, object sorting, and

data output.

3.2.1 Geometric and Order Values

Most of the current software solutions using devices that are integrated with the

printer possess a priori knowledge of dot placement by virtue of being aligned with

the parent device. The positions of the optics and the printer jets are known, and

therefore the expected dot position and actual dot position can be directly coupled.

Error detection subsequently involves comparing centroid coordinates of a given

dot with its expected position.

In cases where the expected and actual positions cannot be directly linked, such as

using a separate optical device not linked to the printer, a different approach must

be taken. Given a m x n dimensional grid of dots, drops are labeled (i ,j ) where i is

the column index, ranging from 1  m, and j is the row index ranging from 1  n

(Figure 3-1). In the ideal case, all dots of index i have the same x coordinate and all

Ryan P. Sullivan 91
dots of index j have the same y coordinate, with an inter-dot spacing d, sometimes

called the pitch.

i=0 𝑥⃗ i=1 i=2 … i=m

j=0
𝑦⃗ θ
j=1

j=2

j=n

Figure 3-1: Layout of grid of m x n dots. In the ideal case 𝑥⃗ and 𝑦⃗ have the same magnitude d and
°
are orthogonal, i.e. θ = 90 .

Two vectors from the centroid of a given dot (i,j) to the centroid of two neighbors,

one in the next row and one in the next column, are defined as:

⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗
𝑋⃑ = (𝑖, 𝑗)(𝑖 + 1, 𝑗) (3-1)

⃗⃗ = ⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗⃗
𝑌 (𝑖, 𝑗)(𝑖, 𝑗 + 1) (3-2)

As the resulting images are decoupled from the original orientation during printing

there will be some degree of rotation introduced, however small it may be. By

taking the Euclidian distance, the length of the line segment connecting two points,

Ryan P. Sullivan 92
and normalizing with the expected inter-dot spacing d we arrive at a dimensionless

value removing any rotation:

‖𝑋⃗‖ (3-3)
Δ𝑥 = ‖𝑋⃗‖⁡⁡⁡⁡⁡⁡⁡⁡⁡Δ𝑥𝑛 ⁡ =
𝑑

⃗⃗‖
‖𝑌 (3-4)
⃗⃗‖⁡⁡⁡⁡⁡⁡⁡⁡⁡Δ𝑦𝑛 ⁡ =
Δ𝑦 = ‖𝑌
𝑑

where an ideal grid will have a value of 1. The further away from a value of 1, the

more unstable a jetting waveform becomes, given pristine, flat substrates. Values

less than 1 indicate contraction in the given X or Y distances, while values greater

than 1 indicate expansion in a given direction. This can be due to change in carriage

velocity, jetting angle, jetting frequency, or in a deviation of impact angle from the

normal. The angle formed between a given drop and its two neighbors is defined

as:

𝑋⃑ ⋅ 𝑌
⃗⃑ (3-5)
𝜃 = cos −1
‖𝑋⃑‖‖𝑌 ⃗⃑ ‖

where a value of 90° indicates an ideal grid. Because these values are dependent

only on the position data of the drops, they do not require a priori knowledge of

intended placement. These values serve to describe the long term stability of

jetting.

Three values are used to describe the morphological properties of individual drops.

The first is the Area, which can be used to determine the volume of deposited

drops given knowledge of the contact angle between a given substrate and the ink.

Ryan P. Sullivan 93
This depends most strongly on the volume of the jet chamber, and to a lesser

extent on the voltage and duration of the waveform. Circularity and roundness

describe how close to an ideal circle the drops are. Higher velocity impacts result in

larger deformation of features, along with splatter, which is undesirable. In cases

where there is a low level of pinning, these values trend toward unity as the drops

return to spherical caps. These values are defined as:

𝑎𝑟𝑒𝑎 (3-6)
𝑐𝑖𝑟𝑐𝑢𝑙𝑎𝑟𝑖𝑡𝑦 = 4𝜋
𝑝𝑒𝑟𝑖𝑚𝑒𝑡𝑒𝑟 2

𝑎𝑟𝑒𝑎 (3-7)
𝑟𝑜𝑢𝑛𝑑𝑛𝑒𝑠𝑠 = 4
𝜋𝑚𝑎𝑗𝑜𝑟𝑎𝑥𝑖𝑠 2

where a circularity of 1 indicates a perfect circle. These values serve to describe the

drop impact and ink-substrate interaction. If the drop has not completely absorbed

the tail before hitting the surface, deformation will be visible. Similarly if the

velocity is too large, the drops will splatter.

3.2.2 Overview of program

3.2.2.1 Calibration Grids

Grids of m x n dots must be deposited on a substrate with an inter-dot spacing d at

least twice the average diameter of the dots. Optical images should be taken so

that the substrates are in the same position as when printed, introducing as little

rotation as possible.

3.2.2.2 Installation and Usage

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The program we have developed is an ImageJ 1.1 plugin that builds on top of the

Otsu Threshold and Analyze Particles plugins. An overview of the general workflow

can be seen in Figure 3-2. It takes in either raw RGB or grayscale images as input

and will attempt to remove uneven illumination of the background and filter out

surface defects. It detects the distribution of Feret lengths, taken here to be the

diameter across the thickest portion of the feature, removes outliers > 2 Standard

Deviations (SD) away from the mean, and performs a rolling paraboloid subtraction

with a radius of x1.5 the max Feret length followed by a 3x3 pixel smoothing matrix.

Once filtered the image is Otsu thresholded and the dots are analyzed; an area filter

is run to further remove extraneous objects. Following this, the remaining features

are sorted into their grid positions and their geometric and order data is obtained.

When run, the user is prompted with a window to input the settings (Figure 3-3).

The software then runs for a couple of hundred microseconds per image before

outputting a results table as shown in Figure 3-4.

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Figure 3-2: Overview of protocol. The manual sections, Data Collection and Data Analysis, are
highlighted in green. The sections that describe the computational processing of the images are
highlighted in blue. Data Collection: Calibration grids are fabricated and imaged. Optionally
alternative segregation methods and manual removal of artifacts may be performed. Input:
Greyscale or binary images along with analysis and output parameters are input. Background
Removal: The software attempts to remove uneven illumination and surface artifacts using Rolling
Paraboloid Subtraction (RPS) from greyscale images based on the distribution of Feret diameters. It
then performs Otsu thresholding resulting in a binary image. User inputted binary images are passed
through. Particle Analysis: Geometric and position values are determined based on the binary image.
Sorting: Features are sorted based on their centroid position analysis parameters. Order values are
determined. Output: The geometric and morphological values for each feature along with analysis
parameters are output to a single table. Binary and Outlined and Labeled images are optional. These
are used to visually verify the output Data in the table. Artifacts can be removed by rerunning with

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altered parameters of manual editing of the binary image which can be input to the Particle Analysis
section directly. Once images of calibration are taken, they can be input as greyscale images. The
software will then threshold the images (Background Removal) and pass the binary image to the
Particle Analysis. Alternatively binary images may be input if users wish to use alternative
thresholding algorithms or need to manually remove artifacts.

Figure 3-3: Input Dialog Window. Background Removal: “None”, “Manual”, “Automatic”, or “Manual
+ Automatic”; determines how the rolling paraboloid subtraction should be performed, if at all.
Thresholding: “None”, or “Otsu”; determines if Otsu Thresholding should be performed - “None”
should only be selected when inputting a binary image. Sorting Method: “Insertion”, “Grid”, or
“Neighbor”; determines the type of sorting method to be used. “Insertion” is the best for high order
grids with no missing features, “Grid” is best for high order grids with missing features, and
“Neighbor” search is for grids of low order and/or missing features. Size (pixel^2): determines the
size range outside of which to ignore features. Column & Rows: determines the dimensions of the
grid the program will search for. Diameter: determines the initial diameter of features looked for
when removing background. Distance: determines the distance between dots; this is used in Grid
and Neighbor sorting methods. Intelligent: determines if the program will automatically filter out
dots based on their area using Pierce’s Criteria when more dots are found than expected. Show
Binary/Outlines/Results/Summary: determines which windows are shown as output. Batch Process:

Ryan P. Sullivan 97
determines whether the program will run on a single image, or iterate through a folder of images. It
will ask for an input and output folder if checked.

As seen in the results output in Figure 3-4 below, there are seven analysis

parameters that are used to describe the ink jetted test patterns. Four of these

parameters, area, perimeter, circularity, and roundness, describe the morphology

of features, and are functions of the droplet velocity, distance until coalescence,

substrate stiffness, and contact angle. The three other parameters, Theta, X, and Y

describe how accurately and reproducibly droplets can be placed at desired

locations.

Figure 3-4: Results Window. The first seven columns contain data of a given feature. Delta X values
are formed with the neighbor on the right; Delta Y values are formed with the neighbor below;

Ryan P. Sullivan 98
Theta values are formed with both neighbors. In the case where a neighbor doesn’t exist, i.e. the
right most column or bottom row, the value is left blank. Columns ten and eleven give cumulative
statistical values for the entire grid along with the parameters used in the processing and analysis.

3.3 RESULTS AND DISCUSSION

3.3.1 Automatic Background Removal

Ideal samples will have pristine substrates and high contrast images; however as is

often the case, data is not always ideal. In cases where there is noise and/or defects

on the substrate or uneven illumination, background removal is necessary for the

Otsu thresholding algorithm to work properly.

Rolling ball background subtraction, first described by Stanley Sternberg (1983), is a

technique used to remove smooth continuous backgrounds from images. Given a

2D image where the intensity is mapped as height, a figurative ball or paraboloid is

rolled along the underside of the surface creating a new background. This

background is then subtracted from the original image, removing low frequency

deviations while leaving the high frequency perturbations intact (Figure 3-5).

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Figure 3-5: Graph of average inverse intensity along the x-axis of a grid on an 8-bit scale before and
after rolling parabaloid removal. Inverse intensity values of 0 are white and values of 255 are black.
In the case of images, the substrate is bright (lower values) and the foreground is dark (high values).
In this case intensity is analogous to height. The unedited images have uneven backgrounds, which
can be seen with rising slope (red). Performing Rolling Parabaloid Removal with a radius of x1.5 the
radius of features removes the uneven background (blue).

As seen in Figure 3-6, Otsu thresholding works equally well with or without

background removal on pristine samples. On noisy/non-ideal samples however,

running Otsu thresholding results in a binary image where large portions of the

background fall into the foreground side of the histogram. When background

removal is performed, the Otsu thresholding has no issues segregating the

foreground from the background.

Ryan P. Sullivan 100

Figure 3-6: Thresholding of Noise Samples. The three images on the left are from a typical pristine
sample, i.e. low levels of substrate defects or artifacts, while the three images on the right are of a
typical noisy sample, i.e. large number of defects or artifacts. The top two images are the unaltered
greyscale images. The middle two images are the results of Otsu Thresholding on unaltered
greyscale images. The bottom two images are the results of Otsu Thresholding on greyscale images
which underwent Rolling Paraboloid Subtraction (RPS). While in the case of pristine samples RPS
makes no significant difference, in the case of noisy samples RPS is vital to proper thresholding.

The diameter of the ball should be at least as large as the diameter of the largest

object not part of the background. The images are processed and the array of Feret

diameters is found. Using a pristine sample, rolling ball and rolling parabola

background subtraction were performed in multiples of the largest Feret diameter,

x1, x1.5, x2 (Figure 3-7). Looking at the effect of the average area of the dots (Table

3-1), the rolling sphere produces results closer to those obtained without any

removal (-0.3% difference) compared to the rolling parabola (-0.6% difference).

However, the rolling ball produces artifacts that become noticeable at x1.5 Feret

Diameters, while the rolling parabola does not. Both have similar reductions on the

standard deviation of areas (-34% for rolling ball, -35% for rolling parabola).

Table 3-1: Validation of Rolling Ball Subtraction. Table contains values derived from the images in
Figure 3-7. The unmodified image was used as the control and was compared to the six background
subtracted images. Error from the Rolling Ball is double at values of x1.5 and x2 compared to x1 the
diameter, but are still under 1% error. The Rolling Paraboloid is even more accurate having 0% error
at x1 and x1.5, and .3% at x2. The standard deviation of both methods is in the low -30% range,
actually reducing the distribution of areas.

None Rolling Ball Rolling Parabola

Radius 1 1.5 2 1 1.5 2
Average 373.8 375.1 375.8 376.1 373.8 373.9 374.9
Relative Change -0.3% -0.5% -0.6% 0.0% 0.0% -0.3%
StDev 9.8 6.2 6.4 6.9 6.3 6.5 6.2
Relative Change -37.1% -34.7% -30.4% -36.1% -34.0% -37.1%

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Figure 3-7: Validation of Rolling Ball Subtraction Parameters. The columns are Rolling Ball
Subtraction and Rolling Parabaloid Subtraction of the same greyscale image (not shown). The rows
are using radii of x1, x1.5, and x2 of the max Feret Radius of the dots in the original greyscale image.
Rolling Ball Subtraction produces strong artifacts once the radius passes the diameter of the dots.
See Table 3-1 for numeric details.

3.3.2 Binarization

To separate the drops from the background, the Otsu Method is used. This

clustering-based thresholding technique takes a greyscale image with a bi-modal

histogram and maximizes their inter-class variance. In an ideal case, a histogram has

a pronounced valley between two peaks representing the foreground and

Ryan P. Sullivan 102

background; the bottom of this valley is the threshold. This produces a binary image

with drops as foreground and the substrate as background (Otsu, 1979).

3.3.3 Sorting

Using the binary image as input, the geometric values area, circularity, and

coordinates of the centroid are calculated for each individual drop using the ImageJ

Plugin Particle Analyzer. The drops are then sorted based on their coordinates using

an insertion sort. Optionally, a binary image, created from separate processing, can

be used.

Sorting can be done in three ways, using Insertion Sort, Grid Sort, or Nearest

Neighbor Sort. Insertion sort works by ordering all the dots by their y coordinate.

The plugin then groups them into rows based on the expected number of columns.

Once split into rows, a second round of insertion sorting is performed based on

their x coordinates (Figure 3-8). It is the simplest and fastest algorithm, but expects

highly ordered grids with no missing features.

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Figure 3-8: Schematic of Insertion Sort Algorithm. First all the dots are ordered by the y-coordinate
of their centroid positions. They are then grouped into rows based on rows and columns parameters.
Each row is then insertion sorted based on the x-coordinate of their centroid positions.

Grid sorting works to create a grid with widths equal to the average inter-dot

distance d. It then bins the dots based on where in the grid they fit by rounding to

the nearest integer. This algorithm requires high order grids, but can take into

account missing features (Figure 3-9).

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Figure 3-9: Schematic of Grid Sort Algorithm. Dots are binned based on where they fit in a grid with
width of d, the inter-dot distance. The plugin can optionally self-correct for drift or deviation from
expected inter-dot distance and reapply the grid based on the experiment average dot distance.

Nearest neighbor sorting works by calculating the positions of where ideal

neighbors should be located, and then compares them to those of the actual

neighbors. All neighbors in an annulus with an outer radius 1.5 d and inner radius of

0.5d are found. They are given a strength factor based on how much they deviate

from the closest ideal position. This then continues for the neighbors of the new

dots until all dots have been fit (Figure 3-10). This algorithm is the most complex,

taking the longest time, but is able to deal with highly disordered systems. Due to

the complexity of the possibility space for highly disordered systems, it is highly

recommended that the results of this algorithm are visually checked to ensure that

they match.

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Figure 3-10: Schematic of Nearest Neighbor Sort Algorithm. Using the top left starting dot, the 8
ideal neighbor positions are calculated based on the input inter-dot distance d. The nearest
neighbors within 0.5-1.5 d are then matched to the ideal positions, and are given a strength value
based on how much they deviate from that position. Found neighbors are then the seed for the next
neighbor and so on.

3.3.4 Validation

While we have discussed the background and efficacy of each of the image

processing steps in sections 3.3.1 to 3.3.3 in isolation, it is important to validate

their combined use. The validity of their use however depends on the

appropriateness of the parameters they are fed given their input images. There are

break points built into the plugin to allow manual adjustment at any given step as

previously seen in Figure 3-2. What is of interest however is the choice of the

parameters the software makes on its own accord. 10,020 dots in 334 grids were

run through the software in automatic mode. That is, given only the input image,

Ryan P. Sullivan 106

the expected number of dots, and the diameter, the software produced all results

without further human intervention. The results for all 10,020 dots were then

manually compared to the original images by overlaying the outline of the dots

resulting from the software automation over the original images to visually

compare likeness. The software was found to accurately identify 9,932/10,020 dots,

giving it an accuracy of 99.12% for feature identification. Of these 88 were

misidentified features, 20 were false positives, 27 were false negative, and 41 were

shifted due the false positives and negatives. Of these shifted 41, 28 occurred in a

single grid that had an extra dot.

Once feature identification was established, feature accuracy was assessed. That is,

whether the geometric values of said features match manual measurements given

the set of identified features. The diameter of the first dot (top left) of the first 100

grids (in alphabetical order of the file names) of the 334 grids used in the previous

test of feature identification was manually measured using optical profilometry

height maps. To minimize the influence of any geometric anisotropy on the

measurements, the diameter of each dot was taken to be the average of

measurements across both the x-axis and the y-axis. The diameter given by the

software was calculated from the area assuming a perfect sphere. These two

diameters were then compared for each of the 100 dots from 100 grids. The

average measured diameter for the population was 23.23 ± 1.60μm compared to

the average 23.50 ± 1.64μm. The software average is 1.17% larger than the

measured average, with a Mean Absolute Percentage Error (MAPE) of 3.00%. It is

Ryan P. Sullivan 107

interesting to note that this MAPE is lower than for x vs y axis measurements, which

have a MAPE of 7.67%. While the manually measured features have high circularity

(between 0.948 ± 0.032 and 0.952 ± 0.007); this difference may be attributed to the

manual measurements serving as only 2 bisections per feature, while the software

effectively tests all possible bisections.

Given an identification error of 0.87% and a measurement error of 3.00%, we have

shown that our software is highly accurate when analyzing features even in

automatic mode, requiring minimal human intervention for analyzing large

datasets.

3.3.5 Example Data Set

Now that we have explained how the program works, example data is shown below

to demonstrate the quantitative output with actual optical images. 12 sets of grids

were printed using identical waveforms, but with different voltages (Figure 3-11).

The top set of grids was printed sequentially at 25 V, while the bottom set of grids

was printed at 30 V. Since the inkjet used prints one line at a time, the first rows

were printed for all six grids, followed by the second rows and so on. The first grid

in each set is highly ordered. As each printing pass continues, instability builds

when using the 25 V parameter; this can be seen visually as the dots start deviating

from their ideal positions in the grids in the top row.

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Figure 3-11: Optical images of inkjetted grids. Each row represents a set of specially aligned grids.
The top row uses parameters of 8 μs - 25 V, 3 kHz while the bottom uses 8 μs – 30 V, 3 kHz. Printing
was performed on a per line basis, e.g. the first row in 1-6, followed by the second row in 1-6, etc.
The images clearly show that while it is easy to obtain a single grid for publication, differences in
parameters change the viability of long term printing. While the 25 V waveform starts breaking
down, the 30V waveform does not (this will be explored in detail in Chapters 4 and 5).

The average θ values for each column from the twelve grids in Figure 3-11 are

graphed in Figure 3-12. While the grids printed with 30 V maintain high θ values,

deviation from 90° can already be seen as early as the first grid of 25 V grids. It

quickly breaks down further as printing continues. While in this example instability

in jetting builds rather quickly, in many cases it builds much more slowly. By having

quantitative data on the grids, degradation in print quality may be detected before

it becomes obvious on visual inspection. This quantitative data allows moving

beyond “good enough” grids, which are seen in many publications, towards the

high reproducibility needed for large scale fabrication.

Ryan P. Sullivan 109

Figure 3-12: Average theta values of each column in the two sets of grids seen in Figure 3-11.

3.4 CONCLUSION

To our knowledge, this is the first open source software implementation for inkjet

pattern analysis. It can be run without the need for proprietary equipment, using

any brightfield microscope and any modern Operating System that supports Java,

eliminating the barrier to entry for labs to start analyzing novel ink jets, inks, and

waveforms. Depending on the applied settings, the software we have developed is

capable of: (i) automatically detecting and removing uneven illumination before

separating features from the background through Otsu Thresholding, (ii) automatic

removal of artifacts such as substrate contamination, (iii) converting the image to

binary, and (iv) sorting the resultant binary image using one of three methods

(insertion sort, grid sort, or nearest neighbor sort), depending on how non-ideal the

grid is expected to be. The sorted data then form the basis for the geometric and

order data which quantify the grids and are automatically output into sorted tables

along with all the settings that were used to perform the analysis. The automation

Ryan P. Sullivan 110

of this software solution significantly increases reproducibility in data analysis as it

removes the issues of user bias and fatigue, and allows users to replicate any and all

steps exactly, and manually change individual parameters in extreme cases.

Furthermore, it allows for the rapid quantitative analysis needed to reach the high

level of reproducibility required in order to fabricate on a large scale.

While the software is currently limited to analyzing a single grid per image, it may

be extended to images, stitched or otherwise, containing multiple grids. When

combined with the Contact Angle and Analyze Stripes plugins (see Chapter 5,

section 5.5.3), this ImageJ plugin provides a complete and automated analysis

solution for rapidly quantifying any inkjet performance as will be demonstrated in

Chapters 4 and 5.

3.5 APPENDIX

3.5.1 Source Code for Inkjet Plugin

Below is the code described in this chapter. Notepad++ was used to color the code

to a custom code set designed to mimic the Fiji macro builder for readability. The

active repository can be found at

https://github.com/RyanPSullivan7/Inkjet_Pattern_Analyzer.

Ryan P. Sullivan 111

 1 /*
2 Inkjet_Pattern_Analyzer is an ImageJ plugin that analyzes optical
3 images of calibration grids
4 and determines the reliability of the printing process.
5 Copyright (C) 2015 Ryan P. Sullivan
6 This program is free software: you can redistribute it and/or modify
7 it under the terms of the GNU General Public License as published by
8 the Free Software Foundation, either version 3 of the License, or
9 at your option) any later version.
10 This program is distributed in the hope that it will be useful,
11 but WITHOUT ANY WARRANTY; without even the implied warranty of
12 MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE. See the
13 GNU General Public License for more details.
14 You should have received a copy of the GNU General Public License
15 along with this program. If not, see
16 <http://www.gnu.org/licenses/>.
17 */
18
19 import ij.*;
20 import ij.io.*;
21 import ij.gui.*;
22 import ij.measure.ResultsTable;
23 import ij.plugin.*;
24 import ij.plugin.filter.BackgroundSubtracter;
25 import ij.process.*;
26 import java.awt.Color;
27 import java.awt.Font;
28 import java.awt.Label;
29 import java.util.ArrayList;
30 import java.util.Arrays;
31 import java.util.List;
32 import java.io.File;
33 import java.io.FilenameFilter;
34 import java.io.IOException;
35 import ij.process.ImageConverter;
36 import ij.util.Tools;
37 import ij.gui.PolygonRoi;
38 import ij.gui.PointRoi;
39
40 public class Inkjet_Pattern_Analyzer implements PlugIn {
41
42 int rows, columns;
43 int countExpected, countActual, countSortedData; //Tracks
44 expected number of features (rows * columns), the found number of
45 features, and sorted number of features + buffers
46 double radius;
47 double distance, ActiveDistance;
48 double deviations = 3;
49 double[][] Data, SortedData, OrderData;
50 protected ImagePlus Image, ActiveImage, BinaryImage;
51 ImageProcessor ActiveImageProcessor;
52 ImageConverter ActiveImageConverter;

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 53 ResultsTable rt, rtPassthrough;
54 double minSize, minSize_Default = 200.0, maxSize,
55 maxSize_Default = Double.POSITIVE_INFINITY;
56 boolean intelligent, binary, outline, showResults, batch;
57 //Variables for dialog window
58 boolean logging = true, Progresslogging = true;
59 String[] BackgroundRemovalMethods =
60 {"None","Manual","Automatic","Manual+Automatic"};
61 String[] ThresholdingMethods = {"None","Otsu"};
62 String[] SortingMethods = {"Insertion","Grid","Neighbor"};
63 String BackgroundRemovalMethod, ThresholdingMethod,
64 SortingMethod;
65
66 String InputDirectory, OutputDirectory;
67 String[] imagelist;
68 String imageName;
69 int filenum;
70
71 //First method called by ImageJ when run. Main() isn't used in
72 ImageJ plugins.
73 public void run(String arg) {
74 if(!showSetup()){
75 return;
76 }
77 if(batch) {
78 setDirectories();
79 for(int i=0; i<filenum; i++) {
80 getImage(i);
81 if(!ProcessImage()){
82 IJ.log("Error: Could not analyze..."+imageName);
83 }
84 else{
85 //CalculateOrder();
86 writeOutputs(i);
87 IJ.log(imageName + " Processed");
88 }
89 }
90 }
91 else {
92 Image=IJ.getImage();
93 imageName = Image.getTitle();
94 reset();
95 if(imageName.endsWith(".tiff")){
96 imageName =
97 imageName.substring(0,imageName.indexOf(".tiff"));
98 }
99 if(imageName.endsWith(".tif")){
100 imageName =
101 imageName.substring(0,imageName.indexOf(".tif"));
102 }
103 if(Progresslogging){

Ryan P. Sullivan 113

104 IJ.log("Working on " +imageName);
105 }
106 if(!ProcessImage()) {
107 IJ.log("Error: Could not analyze");
108 return;
109 }
110 displayOutputs();
111 }
112 IJ.log("Finished");
113 return;
114 }
115 //Setup method that displays a GUI and asks for parameters
116 public boolean showSetup() {
117 GenericDialog gd = new GenericDialog("Grid Analyzer (c) 2015
118 R. Sullivan");
119 gd.addChoice("Background Removal:
120 ",BackgroundRemovalMethods,BackgroundRemovalMethods[0]);
121 gd.addChoice("Thresholding:
122 ",ThresholdingMethods,ThresholdingMethods[1]);
123 gd.addChoice("Sorthing Method:
124 ",SortingMethods,SortingMethods[0]);
125 gd.addStringField("Size (pixel^2):", minSize_Default+"-
126 "+maxSize_Default,10);
127 gd.addNumericField("Colums:",6,0);
128 gd.addNumericField("Rows:",5,0);
129 gd.addNumericField("Diameter",15,0);
130 gd.addNumericField("Distance",80,0);
131 gd.addCheckbox("Intelligent?", true);
132 gd.addCheckbox("Show Binary?", false);
133 gd.addCheckbox("Show Outlines?", true);
134 gd.addCheckbox("Show Results?", true);
135 gd.addCheckbox("Batch Process?", false);
136 gd.showDialog();
137 if (gd.wasCanceled()){
138 return false;
139 }
140 if (gd.invalidNumber()) {
141 IJ.error("Invalid Number!");
142 return false;
143 }
144 BackgroundRemovalMethod = gd.getNextChoice();
145 ThresholdingMethod = gd.getNextChoice();
146 SortingMethod = gd.getNextChoice();
147 columns = (int)gd.getNextNumber();
148 rows = (int)gd.getNextNumber();
149 Double DiameterInput= gd.getNextNumber();
150 distance = gd.getNextNumber();
151 ActiveDistance = distance;
152 //min-max
153 String size = gd.getNextString();

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154 String[] MinMax= Tools.split(size,"-");
155 double min = (MinMax.length >= 1) ?
156 gd.parseDouble(MinMax[0]) : 0.0;
157 double max = (MinMax.length == 2) ?
158 gd.parseDouble(MinMax[1]) : Double.NaN;
159 minSize = min;
160 maxSize = max;
161 //minSize = Double.isNaN(min) ? minSize_Default :
162 mins/unitSquared;
163 //maxSize = Double.isNaN(max) ? maxSize_Default :
164 maxs/unitSquared;
165 //if (minSize<DEFAULT_MIN_SIZE) minSize = DEFAULT_MIN_SIZE;
166 //if (maxSize<minSize) maxSize = maxSize_Default;
167 radius = Math.round(DiameterInput.intValue()/2);
168 countExpected = rows*columns;
169 intelligent = gd.getNextBoolean();
170 binary = gd.getNextBoolean();
171 outline = gd.getNextBoolean();
172 showResults = gd.getNextBoolean();
173 batch = gd.getNextBoolean();
174 return true;
175 }
176 //Prompts user to choose input and output directories, and creates
177 a list of all valid .tiff files in the input directory
178 public void setDirectories() {
179 DirectoryChooser directory = new DirectoryChooser("Choose
180 input directory");
181 InputDirectory = directory.getDirectory();
182 directory = new DirectoryChooser("Choose output directory");
183 OutputDirectory = directory.getDirectory();
184 File folder = new File(InputDirectory);
185 File[] files = folder.listFiles(new FilenameFilter() {
186 @Override
187 public boolean accept(File dir, String name) {
188 //ignore outline.tiff & outline.tif
189 if((name.toLowerCase().endsWith(".tiff") &&
190 !name.toLowerCase().endsWith("outline.tiff"))||
191 (name.toLowerCase().endsWith(".tif") &&
192 !name.toLowerCase().endsWith("outline.tif"))){
193 return true;
194 } else {
195 return false;
196 }
197 }
198 });
199 filenum=files.length;
200 imagelist = new String[filenum];
201 for(int i=0; i<filenum; i++)
202 imagelist[i]=files[i].getName();
203 return;
204 }

Ryan P. Sullivan 115

205 //Reset ActiveImage to current Image
206 public void reset() {
207 ActiveImage=Image.duplicate();
208 ActiveImageProcessor = ActiveImage.getProcessor();
209 ActiveImageConverter = new ImageConverter(ActiveImage);
210 return;
211 }
212 //Sets Image + global parameters based on directory
213 public void getImage(int index) {
214 Opener opener = new Opener();
215 Image = opener.openImage(InputDirectory +
216 imagelist[index]);
217 reset();
218 imageName = Image.getTitle();
219 if(imageName.endsWith(".tiff")){
220 imageName =
221 imageName.substring(0,imageName.indexOf(".tiff"));
222 }
223 if(imageName.endsWith(".tif")){
224 imageName =
225 imageName.substring(0,imageName.indexOf(".tif"));
226 }
227 if(Progresslogging){
228 IJ.log("Working on " + imageName);
229 }
230 return;
231 }
232 //Main method to process image
233 public boolean ProcessImage(){
234 if (BackgroundRemovalMethod=="Manual" ||
235 BackgroundRemovalMethod=="Manual+Automatic"){
236 BackgroundSubtracter subtracter = new
237 BackgroundSubtracter();
238 subtracter.rollingBallBackground(ActiveImageProcessor,
239 radius, false, true,true,false,true);
240 if(Progresslogging)
241 IJ.log("Removed Background with radius " + radius);
242 }
243 if (BackgroundRemovalMethod=="Automatic"||
244 BackgroundRemovalMethod=="Manual+Automatic") {//interative
245 //check if area is * 10 larger than average
246 ActiveImageConverter.convertToGray16();
247 IJ.run(ActiveImage, "Auto Threshold", "method=Otsu
248 white");
249 setMeasurementArray();
250 rtPassthrough = ResultsTable.getResultsTable();
251 int count = StDevFilter(6);
252 IJ.log("Removed " + count + " Feret Outliers, " +
253 countActual + " features left.");
254 radius = CalculateMax(Data[6])*1.5;

Ryan P. Sullivan 116

255 reset();
256 BackgroundSubtracter subtracter = new
257 BackgroundSubtracter();
258 subtracter.rollingBallBackground(ActiveImageProcessor,
259 radius, false, true,true,false,true);
260 if(Progresslogging)
261 IJ.log("Removed Background with radius " + radius);
262 }
263 if(ThresholdingMethod=="Otsu"){
264 ActiveImageConverter.convertToGray16();
265 IJ.run(ActiveImage, "Auto Threshold", "method=Otsu
266 white");
267 //ActiveImage.updateImage();
268 BinaryImage = ActiveImage.duplicate();
269 IJ.log("Thresholded");
270 }
271 setMeasurementArray();
272 //rtPassthrough = ResultsTable.getResultsTable();
273 //check number of expected
274 if(countActual>countExpected) { //if excess number of dots
275 try to filter by area
276 StDevFilter(0);
277 if(countActual>countExpected){
278 IJ.log("Error: Unexpected excess of dots.");
279 return false;
280 }
281 }
282 if(!intelligent) {//Do exactly as user specifices
283 if(SortingMethod=="Insertion"){
284 InsertionSort();
285 }
286 else if(SortingMethod=="Grid"){
287 GridSort();
288 }
289 else{
290 NeighborSort();
291 }
292 }
293 else{//Try and figure optimal sorting
294 if(countActual==countExpected) {
295 if(SortingMethod=="Insertion"){
296 InsertionSort();
297 }
298 else if(SortingMethod=="Grid"){
299 GridSort();
300 }
301 }
302 //consider checking for dots twice the volume
303 else if(countActual < countExpected) {
304 GridSort(); //used for setting distances
305 removeDistanceOutliers();

Ryan P. Sullivan 117

306 if (!GridSort())
307 return false;
308 }
309 }
310 CalculateOrder();
311 return true;
312 }
313 //Display any output windows
314 public void displayOutputs() {
315 if(outline){
316 Outline();
317 ActiveImage.setTitle(imageName + "_outline");
318 ActiveImage.show();
319 }
320 if(binary){
321 BinaryImage.setTitle(imageName + "_binary");
322 BinaryImage.show();
323 }
324 if(showResults){
325 OutputToResults();
326 rt.show("Results");
327 }
328 rtPassthrough.show("Results Passthrough");
329 return;
330 }
331 //Writes any output windows to disk
332 public void writeOutputs(int index) {
333 if(outline){
334 Outline();
335 FileSaver saver = new FileSaver(ActiveImage);
336 saver.saveAsTiff(OutputDirectory+imageName+"_Outline.tiff
337 ");
338 }
339 if(showResults){
340 OutputToResults();
341 try{
342 rt.saveAs(OutputDirectory+imageName+"_Results.csv");
343 }catch(IOException e){
344 IJ.log("Could not write results for "+imageName);
345 }
346 }
347 return;
348 }
349 //Insertion Sort of grid with expected number of dots
350 public void InsertionSort() {
351 countSortedData = countActual;
352 SortedData = Data;
353 for(int i=0; i<rows; i++) {//step by row
354 for(int j=i*columns+1; j<columns*(i+1); j++) {//insertion
355 sort
356 double areatemp = SortedData[0][j];

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357 double xtemp=SortedData[1][j];
358 double ytemp=SortedData[2][j];
359 double perimetertemp = SortedData[3][j];
360 double circularitytemp = SortedData[4][j];
361 double roundnesstemp = SortedData[5][j];
362 double FeretTemp = SortedData[6][j];
363 int k;
364 for(k=j-1; (k>=columns*i) && (SortedData[1][k] >
365 xtemp); k--) {
366 SortedData[0][k+1] = SortedData[0][k];
367 SortedData[1][k+1] = SortedData[1][k];
368 SortedData[2][k+1] = SortedData[2][k];
369 SortedData[3][k+1] = SortedData[3][k];
370 SortedData[4][k+1] = SortedData[4][k];
371 SortedData[5][k+1] = SortedData[5][k];
372 SortedData[6][k+1] = SortedData[6][k];
373 }
374 SortedData[0][k+1]=areatemp;
375 SortedData[1][k+1]=xtemp;
376 SortedData[2][k+1]=ytemp;
377 SortedData[3][k+1]=perimetertemp;
378 SortedData[4][k+1]=circularitytemp;
379 SortedData[5][k+1]=roundnesstemp;
380 SortedData[6][k+1]=FeretTemp;
381 }
382 }
383 if(Progresslogging)
384 IJ.log("Sorted grid using Insertion Sort");
385 return;
386 }
387 //Neighbor Sort of grid
388 public boolean NeighborSort() {
389 if(Progresslogging)
390 IJ.log("Attempting Neighbor Sort...");
391 int[][] RelativeCoordinates = new int[2][countActual];
392 RelativeCoordinates [0][0]=0; //Set initial dot x pos at 0
393 RelativeCoordinates [1][0]=0; //Set initial dot y pos at 0
394 int[][] relativeNeighborPosition = {{1,0},{1,-1},{0,-1},{-
395 1,-1},{-1,0},{-1,1},{0,1},{1,1}};
396 int errorScalar = 2;
397 for(int i =0; i<countActual; i++) {//step through each dot
398 double[][] idealNeighbors = new double[2][8];
399 List<Integer> neighborList = new ArrayList<Integer>();
400 for(int j =0; j<8; j++){//set ideal neighbor positions
401 idealNeighbors[0][j] = Data[1][j] +
402 ActiveDistance*relativeNeighborPosition[j][0];
403 idealNeighbors[1][j] = Data[2][j] +
404 ActiveDistance*relativeNeighborPosition[j][1];
405 }

Ryan P. Sullivan 119

406 for(int j = 0; j<i; j++) {//find all neighbors within
407 distance*errorScalar (2 by default)
408 if(Data[1][j]>Data[1][i]-ActiveDistance*errorScalar &&
409 Data[1][j]<Data[1][i]-ActiveDistance*errorScalar){
410 if(Data[2][j]>Data[2][i]-ActiveDistance*errorScalar
411 && Data[2][j]<Data[2][i]-
412 ActiveDistance*errorScalar){
413 neighborList.add(j);
414 }
415 }
416 }
417 int index = 0;
418 for(int j=0; j<neighborList.size(); j++) {//compare
419 neighbors actual position to ideal neighbors 0,6,7
420 int num = neighborList.get(j);
421 double difference =
422 CalculateDistance(Data[1][num],Data[2][num],idealNeig
423 hbors[0][0],idealNeighbors[1][0]);
424 for(int k=1; k<8; k++) {//compare given neighbor to ideal
425 positions, finding the closest match
426 double euclidianDistance =
427 CalculateDistance(Data[1][num],Data[2][num],idealNei
428 ghbors[0][k],idealNeighbors[1][k]);
429 if(euclidianDistance<difference) {
430 difference = euclidianDistance;
431 index = k;
432 }
433 }
434 //set relative coordinate of new dot
435 RelativeCoordinates[0][j]=RelativeCoordinates[0][index
436 ]+relativeNeighborPosition[index][0];
437 RelativeCoordinates[1][j]=RelativeCoordinates[1][index
438 ]+relativeNeighborPosition[index][1];
439 }
440 }
441 return false;
442 }
443 public boolean GridSort() { //used for initial alignment to use
444 when calculating distances for second run.
445 if(Progresslogging)
446 IJ.log("Attempting Grid Sort...");
447 int[][] RelativeCoordinates = new int[2][countActual];
448 RelativeCoordinates [0][0]=0; //Set initial dot x pos at 0
449 RelativeCoordinates [1][0]=0; //Set initial dot y pos at 0
450 int minXShift = 0; //min relative X position
451 int maxXShift = 0; //max relative X position
452 int minYShift = 0; //min relative Y position
453 int maxYShift = 0; //max relative Y position
454 for(int i =1; i<countActual; i++) { //constant time O(1)
455 double xReal = Data[1][i]-Data[1][0]; //Calculates x-
456 distance given dot and initial dot

Ryan P. Sullivan 120

457 double yReal = Data[2][i]-Data[2][0]; //Calculates y-
458 distance given dot and initial dot
459 int xGrid = (int) Math.round(xReal/ActiveDistance);
460 //Divides by expected distance and rounds to integer
461 int yGrid = (int) Math.round(yReal/ActiveDistance);
462 //Divides by expected distance and rounds to integer
463 RelativeCoordinates[0][i]=xGrid; //Sets x-grid offset for
464 given drop to array
465 RelativeCoordinates[1][i]=yGrid; //Sets y-grid offset for
466 given drop to array
467 if (minXShift > xGrid) minXShift = xGrid;
468 if (maxXShift < xGrid) maxXShift = xGrid;
469 if (minYShift > yGrid) minYShift = yGrid;
470 if (maxYShift < yGrid) maxYShift = yGrid;
471 }
472 int maxXCount = maxXShift-minXShift+1;
473 int maxYCount = maxYShift-minYShift+1;
474 IJ.log("Grid of " + maxXCount + " x " + maxYCount + "
475 formed");
476 if(minXShift < 0 || minYShift <0) {//shift grid so top row,
477 and left column are 0th
478 IJ.log("Shift!");
479 for(int i =0; i<countActual;i++) {
480 if (minXShift < 0){
481 RelativeCoordinates[0][i] =
482 RelativeCoordinates[0][i] - minXShift;
483 }
484 if (minYShift < 0){
485 RelativeCoordinates[1][i] =
486 RelativeCoordinates[1][i] - minYShift;
487 }
488 }
489 }
490 countSortedData = (maxXCount*maxYCount > countActual) ?
491 maxXCount*maxYCount:countActual;
492 SortedData = new double[7][countSortedData];
493 int count=0;
494 for(int i =0; i<countActual; i++) {
495 int position =
496 columns*RelativeCoordinates[1][i]+RelativeCoordinates[0]
497 [i];
498 SortedData[0][position]=Data[0][i]; //Set Area
499 if(SortedData[0][position] !=0){
500 if(count<position){
501 count=position;
502 }
503 }
504 SortedData[1][position]=Data[1][i]; //Set X
505 SortedData[2][position]=Data[2][i]; //Set Y
506 SortedData[3][position]=Data[3][i]; //Set Perimeter

Ryan P. Sullivan 121

507 SortedData[4][position]=Data[4][i]; //Set Circularity
508 SortedData[5][position]=Data[5][i]; //Set Roundness
509 SortedData[6][position]=Data[6][i]; //Set Feret
510 }
511 count++;
512 //countSortedData = count;
513 if(Progresslogging){
514 IJ.log(countActual + " features mapped to " +
515 countSortedData +" places.");
516 }
517 return true;
518 }
519 //finds distances Depricated
520 public void findDistance() {
521 int length=(countActual*(countActual-1))/2;
522 int[] Lengths = new int[length];
523 int count =0;
524 for(int i =0; i < countActual-1; i++) {
525 for(int j = i+1; j<countActual; j++) {
526 int dist =(int) Math.pow(Math.pow(Data[2][j]-
527 Data[2][i],2) + Math.pow(Data[1][j] –
528 Data[1][i],2),0.5);
529 Lengths[count]=dist;
530 count++;
531 }
532 }
533 Arrays.sort(Lengths);
534 for(int i =0;i<count;i++)
535 IJ.log(""+Lengths[i]);
536 }
537 //filters outliers from an array
538 public int StDevFilter(int index) {//need to add interation in
539 cases where count isn't 0
540 if(Progresslogging){
541 if(index == 0)
542 IJ.log("Attempting to remove Area Outliers...");
543 else if(index == 6)
544 IJ.log("Attempting to remove Feret Outliers...");
545 }
546 double[] tempArray = Data[index];
547 int count = 0;
548 double mean = CalculateMean(tempArray);
549 double stDev = CalculateStandardDev(tempArray, mean);
550 double[] tempArrayFilter = new double[countActual];
551 for(int i=0; i<countActual; i++) { //sets up temperorary
552 array 1 or 0 values to act as a mask for the next loop
553 if(Math.abs(tempArray[i]-mean) > stDev*deviations){
554 tempArrayFilter[i]=1;
555 count++;
556 }
557 }

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558 if(count>0){
559 double[][] tempData = new double[7][countActual-count];
560 int j =0;
561 for(int i=0; i<countActual; i++) {
562 if(tempArrayFilter[i] == 0) {
563 tempData[0][j]= Data[0][i];
564 tempData[1][j]= Data[1][i];
565 tempData[2][j]= Data[2][i];
566 tempData[3][j]= Data[3][i];
567 tempData[4][j]= Data[4][i];
568 tempData[5][j]= Data[5][i];
569 tempData[6][j]= Data[6][i];
570 j++;
571 }
572 }
573 Data = new double[7][countActual-count];
574 Data = tempData;
575 }
576 countActual = countActual-count;
577 return count;
578 }
579 //calculates the median value of an array
580 public int CalculateMedian(double[] data){
581 double Median;
582 Arrays.sort(data);
583 int dataCount = data.length;
584 if(dataCount % 2 ==0) {//even
585 int num = (int) (dataCount/2 + .5);
586 Median = (data[num]+data[num+1])/2;
587 }
588 else {
589 Median = data[dataCount/2];
590 }
591 int median = (int) Median;
592 return median;
593 }
594 //uses sortedData to determine distance distribution
595 public void removeDistanceOutliers() {
596 if(Progresslogging)
597 IJ.log("Attempting to remove Distance Outliers...");
598 double[][] DistanceData = new
599 double[2][countSortedData];
600 DistanceData[0]=CalculateX();
601 DistanceData[1]=CalculateY();
602 double[] DistanceData2 = new double[countSortedData*2];
603 System.arraycopy(DistanceData[0],0,DistanceData2,0,countSo
604 rtedData-1);
605 System.arraycopy(DistanceData[1],0,DistanceData2,countSor
606 tedData,countSortedData-1);
607 ActiveDistance = CalculateMean(DistanceData2);

Ryan P. Sullivan 123

608 //DecimalFormat df = new DecimalFormat("#.00");
609 if(Progresslogging)
610 IJ.log("Distance: " + distance + " changed to " +
611 ActiveDistance);
612 return;
613 }
614 //Grabs data from ParticleAnalyzer's result table and sets it as
615 an array
616 public void setMeasurementArray() {
617 if(Progresslogging)
618 IJ.log("Running Analyze Particles...");
619 IJ.run("Set Measurements...", "area center perimeter shape
620 feret's redirect=None decimal=3");
621 IJ.run(ActiveImage, "Analyze Particles...",
622 "size="+minSize+"-"+maxSize+" clear include");//include
623 rt = ResultsTable.getResultsTable();
624 countActual = rt.size();
625 Data = new double[7][countActual];
626 Data[0] = rt.getColumnAsDoubles(0);//set Area
627 Data[1] = rt.getColumnAsDoubles(8);//set X
628 Data[2] = rt.getColumnAsDoubles(9);//set Y
629 Data[3] = rt.getColumnAsDoubles(10); //set Perimeter
630 Data[4] = rt.getColumnAsDoubles(18);//set Circularity
631 Data[5] = rt.getColumnAsDoubles(35);//set Roundness
632 Data[6] = rt.getColumnAsDoubles(19);//set Feret
633 IJ.log(countActual + " features found.");
634 return;
635 }
636 //Outlines and labels current image
637 public void Outline() {
638 if(Progresslogging)
639 IJ.log("Outlining...");
640 IJ.run(ActiveImage, "Outline", "");
641 ActiveImageProcessor = ActiveImage.getProcessor();
642 for(int i=0; i<countSortedData;i++) {
643 if(SortedData[0][i] != 0){
644 String stamp = Integer.toString(i+1);
645 int x = (int)Math.round(SortedData[1][i] –
646 ActiveImageProcessor.getStringWidth(stamp)/2);
647 Font font = ActiveImageProcessor.getFont();
648 int y =
649 (int)Math.round(SortedData[2][i]+ActiveImageProcessor.
650 getFontMetrics().getHeight()/2);
651 ActiveImageProcessor.drawString(stamp,
652 x,y,Color.white);
653 }
654 }
655 }
656 //Writes the final data to Result Table using ImageJ's methods
657 public void OutputToResults() {

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658 if(Progresslogging)
659 IJ.log("Outputing Results..");
660 rt = new ResultsTable();
661 for(int i = 0; i<countSortedData; i++) {
662 rt.incrementCounter();
663 rt.addValue("Area", SortedData[0][i]);
664 if(OrderData[0][i] != 0)
665 rt.addValue("Theta", OrderData[0][i]);
666 else
667 rt.addValue("Theta", "");
668 if(OrderData[1][i] !=0)
669 rt.addValue("Delta X",OrderData[1][i]);
670 else
671 rt.addValue("Delta X", "");
672 if(OrderData[2][i] !=0)
673 rt.addValue("Delta Y",OrderData[2][i]);
674 else
675 rt.addValue("Delta Y", "");
676 rt.addValue("Perimeter",SortedData[3][i]);
677 rt.addValue("Circularity",SortedData[4][i]);
678 rt.addValue("Roundness",SortedData[5][i]);
679 rt.addValue(" "," ");
680 rt.addValue(" ", " ");
681 rt.addValue("Average", " ");
682 rt.addValue("St Dev", " ");
683 }
684 double AreaAverage = CalculateMean(SortedData[0]);
685 double AreaStDev =
686 CalculateStandardDev(SortedData[0],AreaAverage);
687 rt.setValue(8, 0, "Area");
688 rt.setValue(9, 0, AreaAverage);
689 rt.setValue(10, 0, AreaStDev);
690 double ThetaAverage = CalculateMean(OrderData[0]);
691 double ThetaStDev =
692 CalculateStandardDev(OrderData[0],ThetaAverage);
693 rt.setValue(8, 1, "Theta");
694 rt.setValue(9, 1, ThetaAverage);
695 rt.setValue(10, 1, ThetaStDev);
696 double XAverage = CalculateMean(OrderData[1]);
697 double XStDev =
698 CalculateStandardDev(OrderData[1],XAverage);
699 rt.setValue(8, 2, "X");
700 rt.setValue(9, 2, XAverage);
701 rt.setValue(10, 2, XStDev);
702 double YAverage = CalculateMean(OrderData[2]);
703 double YStDev =
704 CalculateStandardDev(OrderData[2],YAverage);
705 rt.setValue(8, 3, "Y");
706 rt.setValue(9, 3, YAverage);

Ryan P. Sullivan 125

707 rt.setValue(10, 3, YStDev);
708 double PerimeterAverage = CalculateMean(SortedData[3]);
709 double PerimeterStDev =
710 CalculateStandardDev(SortedData[3],PerimeterAverage);
711 rt.setValue(8, 4, "Perimeter");
712 rt.setValue(9, 4, PerimeterAverage);
713 rt.setValue(10, 4, PerimeterStDev);
714 double CircularityAverage = CalculateMean(SortedData[4]);
715 double CircularityStDev =
716 CalculateStandardDev(SortedData[4],CircularityAverage);
717 rt.setValue(8, 5, "Circularity");
718 rt.setValue(9, 5, CircularityAverage);
719 rt.setValue(10, 5, CircularityStDev);
720 double RoundnessAverage = CalculateMean(SortedData[5]);
721 double RoundnessStDev =
722 CalculateStandardDev(SortedData[5],RoundnessAverage);
723 rt.setValue(8, 6, "Roundness");
724 rt.setValue(9, 6, RoundnessAverage);
725 rt.setValue(10, 6, RoundnessStDev);
726 }
727 //Calculates the geometric order values and saves them to
728 OrderData array
729 public void CalculateOrder() {
730 if(Progresslogging)
731 IJ.log("Calculating Order Values...");
732 OrderData = new double[3][countSortedData];
733 OrderData[0]=CalculateTheta();
734 OrderData[1]=CalculateX();
735 OrderData[2]=CalculateY();
736 }
737 //Calculates the theta values for a given sorted array of features
738 public double[] CalculateTheta() {
739 double theta[] = new double[countSortedData];
740 for(int i =0; i<rows-1;i++) { //set row
741 for(int j=i*columns;j<(i+1)*columns-1;j++) { //set dot
742 if(SortedData[0][j] != 0 && SortedData[0][j+1] !=0 &&
743 SortedData[0][j+columns] != 0) {
744 double angle1 = Math.atan((SortedData[2][j+1]-
745 SortedData[2][j])/(SortedData[1][j+1]-
746 SortedData[1][j]));
747 double angle2 = Math.atan((SortedData[2][j]-
748 SortedData[2][j+columns])/(SortedData[1][j]-
749 SortedData[1][j+columns]));
750 double result = Math.abs((angle1+angle2)*57.2958);
751 //double check
752 theta[j]= result;
753 }
754 }
755 }
756 return theta;

Ryan P. Sullivan 126

757 }
758 //Calculates the deltaX values for a given sorted array of
759 features
760 public double[] CalculateX() {
761 double x[] = new double[countSortedData];
762 for(int i =0; i<rows;i++) { //set row
763 for(int j=i*columns; j<(i+1)*columns-1;j++) {//set dot
764 if(SortedData[0][j] != 0 && SortedData[0][j+1] != 0)
765 x[j] = SortedData[1][j+1] - SortedData[1][j];
766 }
767 }
768 return x;
769 }
770 //Calculates the deltaY values for a given sorted array of
771 features
772 public double[] CalculateY() {
773 double y[] = new double[countSortedData];
774 for(int i=0;i<rows-1;i++) {//set row
775 for(int j=0; j<(i+1)*columns;j++) {//set dot
776 if(SortedData[0][j] != 0 && SortedData[0][j+columns]
777 !=0)
778 y[j] = SortedData[2][j+columns] - SortedData[2][j];
779 }
780 }
781 return y;
782
783 }
784 //Calculates the Euclidian distance between two points given their
785 coordinates
786 public double CalculateDistance(double x1, double y1,
787 double x2, double y2) {
788 double euclidianDistance = Math.sqrt((y2-y1)*(y2-y1)+(x2-
789 x1)*(x2-x1));
790 return euclidianDistance;
791 }
792 //Calculates the Euclidian distance between two points given their
793 indices
794 public double CalculateDistance(int i, int j){
795 double euclidianDistance = Math.sqrt((Data[1][j]-
796 Data[1][i])*(Data[1][j]-Data[1][i])+(Data[2][j]-
797 Data[2][i])*(Data[2][j]-Data[2][i]));
798 return euclidianDistance;
799 }
800 //Calculates the mean of an array
801 public double CalculateMean(double[] data) { //ignores 0
802 values
803 double sum = 0, mean = 0;
804 int count = 0;
805 for(int i = 0; i<data.length; i++){
806 if(data[i] != 0){
807 sum = sum + data[i];
808 count++;

Ryan P. Sullivan 127

809 }
810 }
811 mean = sum/count;
812 return mean;
813 }
814 //Calculates the standard deviation of an array given the mean
815 public double CalculateStandardDev(double[] data, double
816 mean) { //ignores 0 values
817 double sum = 0, stDev = 0;
818 int count = 0;
819 for(int i = 0; i<data.length; i++) {
820 if(data[i] != 0){
821 sum = sum + Math.pow(data[i]-mean,2);
822 count++;
823 }
824 }
825 stDev = Math.sqrt(sum/count);
826 return stDev;
827 }
828 //Finds the maximum value of an array
829 public double CalculateMax(double[] data) {
830 double max =0;
831 for(int i =0; i<countActual; i++) {
832 if(data[i]>max)
833 max = data[i];
834 }
835 return max;
836 }
837 }

Ryan P. Sullivan 128

 CHAPTER FOUR: 10 PICOLITRE INKJETTING
OF PLATINUM INKS

4.1 INTRODUCTION

Inkjet printing is an important additive fabrication technique seeing many advances

for printed electronics (Fribourg-Blanc et al., 2013; Layani et al., 2012; Reinhold et

al., 2009). It uses active formation and deposition of spheres of ink, instead of

relying on the passive movement of DPN which was explored in Chapter 2. While

inkjet cannot match the resolution of DPN into the nanoscale, it is considerably

faster. For features around 10 μm in size, which is the limit of most piezo Inkjet

printers with 1 pL chambers, a DPN tip must dwell on the order of 2-10 s depending

on the ink, tip, and substrate parameters, not including approach, withdrawal, and

lateral movement before the next feature (O’Connell et al., 2013). Even ignoring the

other movement steps, this results in at best, a rate of 0.1-0.5 Hz, which is five

orders of magnitude slower than the 10,000-15000 Hz of Inkjet printers.

Additionally recent advances in traditional inkjet technology has resulted in Super

Inkjet Technology (SIJ), previously referred to as fine-inkjet, a new commercially

available technique that increases inkjet resolution to submicron levels, ~500 nm

with 0.1 fL droplets (Murata et al., 2005). It is not yet clear what the limit of the rate

of deposition of this technique is.

Ryan P. Sullivan 129

Metals such as silver or copper, in the form of nanoparticle inks, are widely used

due to their relative ease of synthesis, low cost, high conductivity, and low

annealing temperatures (Fribourg-Blanc et al., 2013; Jeong et al., 2008; Zhang et al.,

2014). However these metals are unsuitable for use in medical bionics due to their

degradation and subsequent neurotoxicity. Platinum and Iridium are of particular

interest for their applications in bionic implants and neural electrodes, such as the

Cochlear implant or Argus II retinal implant, due to their long term biostability (Yue

et al., 2016). However it has been considerably harder to produce working platinum

inks and conductive traces, which require higher temperatures or stronger reducing

agents. Nanoparticle inks however may aggregate in print head nozzles, becoming

more problematic as size is scaled down. As recent research has pushed jetting

volumes down from 1-10 pL to 0.1-1 fL (Murata et al., 2005; Sekitani et al., 2008),

particle size and aggregation have become much more important as they should

generally be 1/100th of the size of the nozzle (Cummins and Desmulliez, 2012).

Furthermore the stabilizing agents used to prevent this precipitation and

agglomeration can themselves impede sintering of the printed nanoparticles,

creating insulating layers (Wu et al., 2007). While recent work has been done to

extend organometallic inks for platinum, removing clogging issues, these still

require high temperatures (350°C) to reduce the inks (Cummins et al., 2011; Goldie

et al., 2014).

In this Chapter we continue the work from Chapter 2, and our previous work from

O’Connell (2013) on polyol inks, in which a metal salt (H2PtCl6) is dissolved in a

polyol (ethylene glycol), which serves both as reducing agent and ink carrier. As

Ryan P. Sullivan 130

polyol inks do not contain any particles which could aggregate, the clogging of

smaller jets becomes a non-issue, allowing scalability down to the levels of SJT in

the future. To our knowledge, this is the first case of using polyol inks in inkjet

printing. The printing of conductive inks is more demanding than other inks, as even

small deviations or errors in printing have the potential to cause breaks and shorts

in the resulting electronics (Cummins and Desmulliez, 2012). As such we have used

the software developed in Chapter 3 to rigorously evaluate jetting waveforms.

Using medical grade PDMS similar to that used in commercial cochlear and retinal

implants as the substrate, the jetting of polyol inks has also been studied and

optimized for maximum stability and firing rate in this Chapter. Print lifetime, the

allowable time after deposition of the first feature before subsequent treatments

must be performed, has also been analyzed through conductivity, volume, and

elemental characterization of deposited features.

4.2 EXPERIMENTAL DETAILS

4.2.1 Ink Formulation

Polyol inks were created by dissolving chloroplatinic acid hydrate (H2PtCl6 (520896

Aldrich) in ethylene glycol (EG). Inks were made as outlined in Table 4-1, to produce

1 g of ink based on the desired metal loading. The inks in Table 4-1 have been

expressed in terms of Pt weight instead of metal salt weight in order to make it

directly comparible to loading of nanoparticle inks (see Appendix 4.5.1). A 10%

loading was chosen for this study over the higher loadings (25%) used in Chapter 2

and our previous work (O’Connell et al., 2013) in order to: (i) lower costs of large

Ryan P. Sullivan 131

scale testing, and (ii) extend the lifetime of the deposited ink, by increasing the time

before there is insufficient EG to complete the reaction due to evaporation.

Table 4-1: Metal loading lookup table for ink formulations based on desired Pt content. Masses are
such that the total weight of the ink is 1g so that the table can easily be scaled. Each column
represents a given metal loading in 5% increments to allow for comparisons with other metal inks.
The weights of platinum salt and ethylene glycol (EG) for 1ml of ink are given in grams. The vapor
pressure of each ink is given in Pascals. The stochiometric ratios are given as EG: H2PtCl6. See
Appendix 4.5.1 for calculations.

Loading 5% 10% 15% 20% 25% 30% 35% 40% 45%

H2PtCl6 (g) 0.11 0.21 0.32 0.42 0.53 0.63 0.74 0.84 0.95
EG (g) 0.89 0.79 0.68 0.58 0.47 0.37 0.26 0.16 0.05
Pvap (Pa) 7.37 7.21 7.01 6.76 6.43 5.96 5.28 4.18 2.08
Ratio 56.26 24.83 14.35 9.11 5.97 3.87 2.38 1.26 0.38

4.2.2 In Flight Jetting

Ink Jetting was performed with a Diamatix Materials Printer (DMP-2800 Series). 1.5

mL of ink was loaded into Diamatix cartridges and jetted using 10 pL jets. The

waveforms used can be seen in Figure 4-1. The voltage and duration of each

segment were adjusted until quasi-stable jetting was achieved. Ink jets require a

negative pressure to keep the meniscus in place, so the typical vacuum of 4.0 inches

H2O was used (FUJIFILM Dimatix, Inc., 2010). Using the printer’s built in optics,

images and video were captured of drops in flight at the exit of the jets. These

images are composites of many different drops using a μsec flash bulb. Velocity,

shape, size, and time until coalescence were measured as functions of applied

voltage and waveform duration to optimize the waveform.

Ryan P. Sullivan 132

Figure 4-1: Five phases of inkjet stimulation waveform. 0: Baseline; 1: Contraction of piezo and
expansion and filling of chamber; 2: Expansion of piezo, contraction of chamber, expulsion of ink; 3:
Partial expansion of pizeo, contraction of chamber to “nip” the ink jet; 4: Return to baseline.

4.2.3 PDMS Substrate Creation

As described previously in Chapter 2, glass slides were etched using a diamond

tipped pen, and then broken into 1 cm x 0.5 cm pieces. They were then cleaned

using filtered water and sonicated for 10min, then dried using N2 stream.

The following is a time-sensitive protocol and was performed exactly in the order

described.

Glass petri dishes (3 cm in diameter) were cleaned, and immersed in water with

Palmolive Original Dishwashing Liquid (Colgate-Palmolive, USA) for 1 hr to allow a

surfactant film to build on the glass. This was necessary to allow easy removal of

final PDMS product. The water was drained from each petri dish and the tip of a

Kimwipe (Kimtech Science*, USA) was used to absorb the small amount of

remaining water in the dishes and rupture any bubbles that had formed. The dishes

were then air-dried for a further 2 hr.

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1 g MED-4234 Part A (NuSil, USA) was added to a glass vial with screw top lid, and

dissolved in 10 mL in n-hexane by alternating between 30 s of vortexing and manual

stirring with a metal spatula until there completely dissolved. The solution was

subsequently sonicated for 30 s to remove any dissolved gases.

Glass pieces were cleaned by rinsing and 10 min sonication with water, following by

rinsing and 10 min sonication with IPA, followed by drying under N2 stream.

Cleaned glass pieces were placed in a Plasma Cleaner (PDC-002, Harrick, USA) and

were cleaned using atmospheric plasma at 30 W for 10 min, allowing the creation

of additional hydroxyl groups on the glass to which the PDMS can bind. Glass pieces

were then placed in the dry petri dishes in a fume hood in preparation for the

PDMS solution to be poured.

0.1 g of MED-4234 Part B was added to the MED-4234 Part A/n-hexane solution.

The solution was manually swirled for 10 s to ensure it thoroughly mixed. The

resulting solution was then poured into the petri dishes in the fume hood to a

depth of 1cm, completely covering the prepared glass slides. The solution was left

to evaporate in the fume hood overnight.

The following morning the petri dishes, now coated with a thin film of uncured

PDMS, were placed in a thermal oven at 120°C for 3 hr. The petri dishes were

removed, and left to cool to room temperature. Once cooled, a scalpel blade was

used to cut the now cured PDMS film along the edges of the glass pieces to allow

the removal of the glass-PDMS substrates. Each substrate was marked at one end,

Ryan P. Sullivan 134

cleaned under a stream of distilled water for 10 s, dried with N 2, and then placed in

individual cleaned 1.5 mL Eppendorf tubes for storage until required.

4.2.4 Ink Jetting of Grids

Grids of 6 x 5 dots were printed on PDMS substrates using the 10 pL jets. Total

pulse duration, applied voltage, and firing frequency of the waveform were altered

over ranges of 8-12 μs, 25-40 V, and 1-15 kHz respectively. A single printed 6 x 5

grid was used as the unit for a larger 6 x 6 grid. Each unit grid was then imaged with

a x25 magnification objective on a Wyko NT9100 Optical Profilometer (Veeco, USA).

The resulting images were then processed using the ImageJ plugin described in

Chapter 3. Results were then compared using Mann-Whitney U tests in SPSS to

ascertain statistical significance; the level of significance was set to P<0.01.

4.2.5 Reduction of H2PtCl6

Using the optimized waveform determined in section 4.2.4, which will be discussed

in results section 4.3.2 below, grids of 3 x 2 were printed onto PDMS and Gold-

Mylar. The smaller grid size was chosen due to easier imaging at higher

magnifications for higher resolution height maps. The following methods were

explored for reduction of printed metal inks.

For plasma reduction, samples were placed in a petri dish and placed in the back of

the plasma chamber. The chamber was sealed and the vacuum started, with inlets

closed. When a pressure of 500 mTorr was reached, the chamber was flooded with

Ryan P. Sullivan 135

N2 until a pressure of 1050 mTorr was reached. The plasma chamber was then

turned on at either 5 W or 30 W power setting for durations between 0.5-90 min.

For microwave reduction, samples were placed in a petri dish and stimulated using

a 1000 W microwave for 1, 2, or 3 min.

For thermal reduction, samples were placed in a thermal oven at 200 °C for 90 min.

Samples were measured using the Optical Profilometer immediately before and

after each reduction treatment to capture changes in the morphology of the

printed features.

4.2.6 Conductive AFM

Using an Asylum AFM (MFP-3D Classic, Asylum Research, USA) in ORCA mode with

a conductive cantilever tip, height and conductivity images of the printed features

were obtained. Point IV curves were taken at nine different locations per image

(Figure 4-2), four in contact with the Au substrate at the corners and five on the Pt

feature. Each point was taken at 5 different applied forces to account for

differences in contact area. Additional point IV curves were taken at places of

interest.

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Figure 4-2: Schematic representation of C-AFM measurements. Orange-red is Gold-Mylar substrate.
Orange-yellow is Pt feature. Blue circles are IV locations.

4.2.7 SEM-EDS

SEM images and EDS spectra were obtained using a JOEL SEM-6490LA using an

acceleration voltage of 15 kV and an energy range of 0-20 keV. Due to the low

vacuum nature of the SEM (40 Pa), no modification of the substrates was necessary.

4.3 RESULTS AND DISCUSSION

4.3.1 In Flight Jetting Analysis

4.3.1.1 Drop Volume

Captured images were found to be low contrast with high background noise (Figure

4-3). Drops and the area immediately around them were cropped and Otsu

thresholding performed to separate the drop from the background. Failure to crop

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would often cause background noise to be registered as foreground. While subpixel

edge detection would be the preferred method for analysis, the resolution was not

high enough.

Figure 4-3: Representative Images of droplets (10 μs pulse) at various voltages (22-27 V) with a head
traveling downwards. Images were taken just above the jet, at the same height. Larger voltages
caused larger velocities, which can be seen in the increased distance travelled. At a certain point
two spheres are created which will not coalesce.

A falling droplet with or without a tail can be separated into multiple circular slices

(Figure 4-4) orthogonal to the velocity vector (van der Bos, 2010; van der Bos et al.,

2014).

Figure 4-4: Schematic of a droplet with a tail. A given slice (black), has a thickness of

xk and a diameter of yk

The volume can be obtained by summation over the areas of all the slices:

𝑛
(4-1)
𝑉 = ∑ 𝐴(𝑥𝑘 )𝑥𝑘
𝑘=1

In the case of using single pixel thick slices:

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 𝑦𝑘 2 (4-2)
𝐴(𝑥𝑘 ) = 𝜋 ( )
2

Using slices a single pixel thick,⁡𝑥𝑘 = 1, the equation then becomes:

𝑛
𝜋 (4-3)
𝑉 = ∑ 𝑦𝑘 2
4
𝑘=1

The resulting volumes shown in Table 4-2 were calculated using equation 4-3 by

manually measuring the pixels in each row and summating.

Table 4-2: Average volume data obtained by summating circular slices perpendicular to the travel
vector of droplets in flight with 3 individual ink jets. Lack of standard deviations indicates that only a
single jet produced quantifiable data.

20V 21V 22V 23V 24V 25V

8 μs 7.1 ± 1.0 pL 7.8 ± 1.0 pL 12.2 ± 5.7 pL 12.9 ± 4.3 pL 28.1 ± 22.2 pL 35.7 ± 21.0 pL
9 μs 7.9 ± 1.0 pL 7.2 ± 4.2 pL 7.8 ± 3.9 pL 7.4 ± 3.2 pL 7.0 ± 1.7 pL 11.2 ± 5.1 pL
10 μs 5.3 ± 1.0 pL 5.6 ± 0.7 pL 6.1 ± 0.5 pL 6.3 ± 0.4 pL 7.2 ± 0.4 pL 5.6 ± 0.4 pL
11 μs 8.0 pL 9.1 pL 8.6 pL 8.8 pL 8.3 pL 18.3 pL
12 μs 6.6 pL 7.0 pL 6.9 pL 10.7 pL 16.1 pL 17.2 pl

Table 4-2 shows an increasing trend with voltage as expected (FUJIFILM Dimatix,

Inc., 2010), though there is large variation. The effect of pulse duration is less clear.

A positive relationship would be expected, but a U shaped trend is seen, with 10 μs

having the smallest volumes at a given voltage, with volume increasing both

towards 8 and 12 μs. This is likely due in part to high levels of variation in the

readings obtained. It has previously been determined that refraction, droplet

flattening, and fluid oscillation may lead to volume errors up to 10% in an ideal

system (Cummins and Desmulliez, 2012).

This high level of variability in readings may be due to three factors; (i) the low

magnification (x4) of the optics used to capture the image. x20 is what is commonly

Ryan P. Sullivan 139

used in the literature (van der Bos et al., 2014; O’Connell et al., 2013), (ii) the low

resolution of the optics, 2.63 μm/pixel, relative to droplet cross-sectional area

0.365 μm/pixel (van der Bos, 2010; van der Bos et al., 2014), and (iii) the high levels

of background noise in the captured frames. The diameters of the slices range from

~1-15 pixels, which is equivalent to 2.6-39.5 μm. Due to the small size of the

droplets and high background noise of the images, any error in thresholding from

true values is extremely large. Assuming the edge of droplets separated from

background is off by a single pixel per side of the droplet, the error in slice area

ranges from 900% to -200% for a single pixel, down to 128% to 75% at 15 pixel

diameter (Figure 4-S1). While summation over slices allows calculation of volume

for droplets with tails, the small diameter of tails on the order of 1-3 pixels,

exacerbates the error due to the poor binning.

Over the course of a printing session, the jet shifts between “modes.” Modes are

stable, ideal jetting, or quasi-stable jetting regimes, modes caused by dampening.

At higher frequencies, there are smaller delays between waveforms. If the delay is

shorter than the time it takes for the liquid to return to resting state, perturbations

may build up damping subsequent waveforms, resulting in lower volumes and

droplet velocities. These may also cause changes in jetting angle and even the

number of droplets formed (in cases where two smaller droplets are ejected

instead of one). At higher frequencies and during longer jetting sessions, instability

builds over time. The default or true mode is taken to be the most common; largest

stable jet immediately after cleaning and preparing the jets. In the case of more

stable jets, there is no apparent change in jetting. Droplets which did not form

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properly under the given parameters were not plotted (Figures 4-5 and 4-6), as

seen with 10 μs – 20 V and 10 μs – 21 V.

4.3.1.2 Drop Velocity

Velocities were calculated by measuring the distance traveled between the

different strobe delays. The velocities obtained show a positive linear relationship

with the applied voltage (Figure 4-5). The relationship between pulse width and

velocity is not as clear. Due to the high variation it is hard to see significant trends;

while 10, 11, and 12 μs pulse durations show increasing velocity with voltage as

would be expected, 8 and 9 μs do not clearly show this. We attribute this to the low

magnification, high background noise, and higher instabilities in 8us pulses. While it

could not be readily captured in tables, 8us pulses tended to have relatively short

stable jetting lifetimes on the order of 10's of seconds before printing would

become unviable.

Figure 4-5: Velocities of the droplets as a function of the applied voltage (V) and pulse length (μs).
There is a clear positive linear relationship for voltage.

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4.3.1.3 Drop Coalescence

The distance until coalescence is the distance it takes for a droplet with a tail to

form a single spherical droplet. This provides a minimum working distance between

the inkjet head and substrate. If the inkjet head and the substrate are closer than

the minimum working distance, then the jetted pattern will be more chaotic as the

tail will cause adverse impact interactions. The distance until coalescence was

found to increase strongly with voltage, but generally lowered with increasing pulse

durations (Figure 4-6).

Figure 4-6: Distance until sphere is the distance the droplet needs to travel before forming a perfect
sphere. If the distance between the jet and the substrate is shorter than this distance, the droplet
will impact with the substrate before it forms a sphere, resulting in chaotic impacts (Rioboo et al.,
2002). This distance is determined by altering the strobe delay until the droplet has travelled
sufficiently far to form a complete sphere. Droplets that traveled outside the field of view before
completely coalescing could not be recorded.

Ryan P. Sullivan 142

4.3.2 Inkjetted pattern analysis

As an alternative to in-flight volume and jetting angle analysis and as a complement

to velocity and jet formation analysis, feature geometry and pattern characteristics

were analyzed. Large scale fabrication needs to both be able to reproducibly

produce droplets of identical size, the morphology, and reproducibly place them at

desired location, the order. As discussed in detail in Chapter 3, the morphology of

droplets is characterized by their area, circularity, and roundness. An ideal droplet

will be a perfect spherical cap, with minimized area, and circularity and roundness

values approaching 1. For this to be the case, the droplets must completely

coalesce before hitting the substrate. Additionally, overly high droplet velocities

may cause the droplets to splatter when they hit the substrate, as all momentum

must be absorbed by inelastic collisions. Also discussed in detail in Chapter 3, the

order of droplet jetting is measured by theta, the angle formed by a droplet and its

two neighbors, ideally a right angle, and the vertical and horizontal pitch.

Inkjet droplet vectors are the combination of the ejection velocity, described above

in 4.3.1.2 with a stationary carriage, and carriage velocity, which will be non-zero

during printing. Alteration of either source caused errors in the resulting grids.

Variation in droplet ejection velocity caused in-line shifts of droplet position, i.e.

acute dampening of droplet ejection caused increases in x-axis distance between

droplets. In general printing can be classified into four main phases. The first is

when the jetting is unstable, producing sprays and features of variable size and

position (Figure 4-7A). The second is when droplet formation has stabilized, and all

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features are of roughly equivalent volume, but there are still errors in positioning

(Figure 4-7B). The third phase is when volume and positioning are stabilized

producing highly ordered grids (Figure 4-7C). The last phase is optimization to

minimize the volume and maximize the jetting frequency (Figure 4-7D).

A B

C D

Figure 4-7: Representative optical images of unreduced inkjetted features on PDMS that represent
the four main phases of jetting optimization. A: unstable volume and positioning; B: stable volume
and unstable positioning; C: stable volume and positioning. D: stable volume and positioning, with
minimized volume optimization. Scale bar: 100 μm.

The collated data obtained from images of the grids over a range of durations of 8-

12 μs and a range of voltages of 25-40V can be found in Tables 4-3 to 4-6. Any grids

or features that exhibited leaks or sprays were ignored and printing was stopped to

purge the nozzles of the cartridges. Data shown is from six rows of six grids (6 x 6),

with each unit grid having thirty (6 x 5) features. Averages are derived from these

1080 features per parameter combination, minus any removed due to artifacts. In

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terms of area minimization, lower voltages and pulse durations produce the

smallest features (Table 4-3). Average diameters were determined from area values

assuming perfect circles, which is close given the roundness values (Table 4-5).

Volumes were then calculated from the radii of the features using the formula for

volume of a spherical cap:

𝜋𝑟 3 (4-4)
𝑉= (2 − 3 𝑐𝑜𝑠 𝜃 + 𝑐𝑜𝑠 𝜃 3 )
3

(Winkler et al., 2014). Using the contact angle of 102° yields:

𝑉 = 1.776𝑟 3 (4-5)

The resulting ranges in feature size are 19.73 ± 5.28 to 27.11 ± 9.49 μm for

diameter, 305.49 ± 21.87 to 576.77 ± 70.66 μm2 for area, and 1.70 ± 0.03 to 4.42 ±

0.19 pL for volume. Relative to the smallest feature, this represents a 37% range for

diameter, an 89% range for area, and a 160% range for volume (Table 4-3). Figure

4-8 shows four grids printed with 8 μs pulse at voltages of 25, 30, 35, and 40V

demonstrating increasing size with voltage.

Figure 4-8: Effect of voltage on feature size. All 4 images use an 8 μs pulse with voltages of 25, 30,
35, and 40V respectively.

Ryan P. Sullivan 145

Table 4-3: Diameter, area, and volume averages of features using jets at 3 kHz firing rate. Distance
and volume values were derived from the area data using the roundness and contact angle
relationships.

25 V 30 V 35 V 40 V
19.73 ± 5.28 μm 21.26 ± 4.96 μm 23.24 ± 6.76 μm 24.46 ± 4.82 μm
8 μs 305.49 ± 21.87 μm2 354.97 ± 19.29 μm2 423.99 ± 35.92 μm2 469.60 ± 18.25 μm2
1.70 ± 0.03 pL 2.13 ± 0.03 pL 2.79 ± 0.07 pL 3.25 ± 0.02 pL
21.51 ± 3.79 μm 22.89 ± 4.79 μm 25.56 ± 6.62 μm 26.87 ± 6.37 μm
10 μs 363.04 ± 11.25 μm 411.41 ± 17.98 μm 512.98 ± 34.38 μm 566.93 ± 31.81 μm2
2 2 2

2.21 ± 0.01 pL 2.66 ± 0.02 pL 3.71 ± 0.06 pL 4.31 ± 0.06 pL

21.31 ± 4.80 μm 23.91 ± 3.88 μm 24.22 ± 7.21 μm 27.11 ± 9.49 μm
12 μs 356.60 ± 18.08 μm2 448.71 ± 11.83 μm2 460.40 ± 40.85 μm2 576.77 ± 70.66 μm2
2.15 ± 0.02 pL 3.03 ± 0.01 pL 3.15 ± 0.08 pL 4.42 ± 0.19 pL

While the original intent of this study was to minimize the size of the printed

features, this was found to come at a significant cost to the stability of the features,

measured by theta. Therefore, stability, which is defined as having theta values

closest to 90°, was chosen as the first criteria in choosing a set of parameters to use

as the final waveform (Table 4-4). The parameters 8 μs – 30 V, 8 μs – 40 V, and 10

μs – 30 V were found to be the most stable and had statistically undistinguishable

means (p = 0.052 and 0.114) from one another; however these parameters were

found to be significantly better than all other parameters, with the highest p-value

being p = 0.005 with all other values being p < 2.04*10-9. Of these three sets of

parameters, 8 μs – 30 V was eliminated due to higher standard deviation of theta

compared to 8 μs – 40 V and 10 μs – 30 V, ± 3.8 vs 1.0 and 0.9 respectively. Of the

remaining two sets of parameters of equal maximized stability statistically speaking,

the size of deposited droplets was chosen as the next limiting factor. 10 μs – 30 V

was chosen for having smaller features than 8 μs – 40 V, with diameters of 22.89 ±

4.79 vs 24.46 ± 4.82 μm (p = 1.06*10-204) (Table 4-4).

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Table 4-4: Average theta values of grids using jets at 3 kHz firing rate. Theta is the angle formed
°
between a given drop and two neighbors; in a perfectly placed grid, all theta values would be 90 .
°
Deviation from 90 therefore indicates lower reproducibility in droplet placement.

25 V 30 V 35 V 40 V
8 μs 78.8 ± 16.5° 89.1 ± 3.8° 88.9 ± 4.9° 89.2 ± 0.9°
° ° ° °
10 μs 88.6 ± 1.4 89.1 ± 1.0 87.9 ± 1.6 87.9 ± 1.7
12 μs 88.3 ± 2.1° 88.4 ± 1.7° 87.7 ± 1.7° 87.8 ± 2.6°

There was no strong trend in the circularity and roundness values of the printed

features (Table 4-5). The momentum of droplets must be dissipated upon impact,

usually in the form of inelastic collisions or droplet bouncing. If the kinetic energy is

too high, as could be the case with high velocities, the ink may spread further or

even splatter. The granularity of the perimeters (on the order of 80 pixels) and the

high contact angle of the ink on the PDMS substrate may however mask any such

effects. The latter causes deposited ink to form ideal spherical caps after impact

with no pinning, thus masking any possible issues from higher velocities resulting

from higher applied voltages. It will remain to be seen however if this effect

changes on stiffer and more hydrophilic substrates, such as platinum, as may be the

case in multi-layer deposition in future experiments.

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Table 4-5: Average circularity and roundness of printed features using jets at 3 kHz firing rate.

25 V 30 V 35 V 40 V
0.946 ± 0.020 0.942 ± 0.034 0.933 ± 0.076 0.933 ± 0.021
8 μs
0.946 ± 0.006 0.948 ± 0.032 0.946 ± 0.076 0.952 ± 0.007
0.939 ± 0.015 0.941 ± 0.012 0.932 ± 0.035 0.931 ± 0.026
10 μs
0.948 ± 0.006 0.952 ± 0.005 0.954 ± 0.030 0.956 ± 0.008
0.942 ± 0.018 0.936 ± 0.013 0.922 ± 0.027 0.934 ± 0.065
12 μs
0.949 ± 0.007 0.952 ± 0.005 0.949 ± 0.009 0.953 ± 0.048

The X and Y distances (discussed in section 3.2.1 in the previous chapter) show no

strong trend outside large deviations at 8 μs – 25 V. These distances are typically

more useful for showing alterations in carriage or jetting velocities, in which case X

distances will be stretched or compressed compared to the Y values. Changes in

carriage velocity usually show at the beginning and end of horizontal segments if

the printer’s motor control doesn’t take into account acceleration and deceleration

respectively. In the case of jetting velocity, bubbles in the line along with waveform

instabilities can dampen the velocity. While these were seen while developing

waveforms, none of this is present in the parameters presented.

Table 4-6: Average X and Y distances between features in grids jets at 3kHz firing rate. The set
distance between features was 80 μm.

25 V 30 V 35 V 40 V
86.6 ± 15.3 μm 79.9 ± 3.4 μm 80.3 ± 7.0 μm 79.8 ± 2.0 μm
8 μs
81.7 ± 22.8 μm 78.9 ± 4.4 μm 78.9 ± 4.3 μm 79.0 ± 4.1 μm
80.4 ± 2.1 μm 79.9 ± 2.2 μm 79.9 ± 2.3 μm 79.9 ± 2.6 μm
10 μs
78.5 ± 3.7 μm 79.0 ± 2.8 μm 79.4 ± 2.7 μm 79.1 ± 3.9 μm
80.0 ± 2.7 μm 79.7 ± 2.6 μm 79.8 ± 2.4 μm 79.8 ± 3.2 μm
12 μs
79.5 ± 3.7 μm 78.4 ± 2.8 μm 78.4 ± 3.7 μm 78.8 ± 3.8 μm

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It is important to note that since the data is averaged over 1000s of features, any

data on the time variance in the instability is lost. The parameters of 8 μs – 25V

produce a striking example of the time variance of the instability. Looking at the top

row in Figure 4-9, it can be seen how with each subsequent dot printed in a row,

jetting becomes more unstable. The parameters of 8 μs – 30V however (bottom

row in Figure 4-9), do not show strong time variance. Ultimately the 10 μs – 30 V

parameters were chosen for having high stability, i.e. theta values closest to 90°

with minimal spread, smaller features, and lower velocities, which becomes more

important on harder, less hydrophobic surfaces (Rioboo et al., 2002).

Figure 4-9: Optical images of inkjetted grids. Each row represents a set of specially aligned grids. The
top row uses parameters of 8 μs – 25 V, 3 kHz while the bottom uses 8 μs – 30 V, 3 kHz. Printing was
performed on a per line basis, e.g. the first row in 1-6, followed by the second row in 1-6, etc. The
images clearly show that while it is easy to obtain a single grid for publication, differences in
parameters change the viability of long term printing. While the 25 V waveform starts breaking
down, the 30 V waveform does not.

4.3.3 Morphology of Features based on Reduction

Data were extracted from Optical Profilometry height images of x50 magnification,

using Gwyddion. Images were plane flattened and 10 pixel (9.76 μm) wide manual

traces over the center of features were taken and the distance from peak to trough

was taken as the height of the features (Table 4-7).

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Table 4-7: Average width and height of features after the different treatments represented as a
percentage of before treatment width and height. The number in brackets is the number of minutes
that each of the different treatments was applied. Vacuum without plasma was used as a control to
separate the effects of volume loss to due ink evaporation in the vacuum and volume loss due to
interaction with the plasma. As can be seen, an 11% drop in feature height can be achieved with
only evaporation under vacuum (1100 mTorr) for 10 min, while plasma treatment for the same time
produces a 47% drop in feature height. Continued duration of high wattage plasmas have been
shown to condense metal films (Loo et al., 2002). While there is a downward trend from 10 to 120
min of continuous plasma treatment, the high standard deviation precludes conclusive results.

Treatment Width Height

Vacuum Only (10 min) 101.53 ± 5.97% 89.05 ± 3.68%
Vacuum + Plasma (10 min) 92.66 ± 5.93% 52.24 ± 10.28%
Vacuum + Plasma (25 min) 96.52 ± 7.38% 55.82 ± 18.38%
Vacuum + Plasma (60 min) 100.15 ± 4.10% 47.34 ± 8.66%
Vacuum + Plasma (120 min) 100.34 ± -55.30% 44.7 ± 7.26%
Thermal 200°C (90 min) 98.38 ± 7.55% 40.62 ± 11.61%

Changes in the width of the features are difficult to discern due to the rupturing

nature of the features under plasma treatment. A small layer of ink spreads along

the surface of the PDMS, while the bulk of the ink stays confined to its initial

location before it ruptures (Figure 4-10C, E). We attribute this to the modification of

the PDMS with hydroxyl groups, increasing its hydrophilicity, thereby changing the

contact angle of the features. Vacuum treatments alone, where the sample is put in

the plasma chamber under the normal protocol except that the plasma is not

turned on, do not produce these effects, supporting the hypothesis of plasma as

the cause (Figure 4-10B). Vacuum treated samples have the same morphology as

immediately after deposition, except for lower heights due to evaporation (Table 4-

7, Figure 4-10A,B)

While there was an initial drop in height by roughly half by the 10 min time-point,

there was no further statistically valid drop (Table 4-7). It is known that reduction

Ryan P. Sullivan 150

takes place at the liquid-plasma interface (Koo et al., 2005, 2006; Shim et al., 2007;

Wang et al., 2009). Platinum nanoparticles form by the polyol process due to

plasma stimulation. If the density of formed nanoparticles is high enough, the

plasma may sinter them together by removing the attached EG byproducts and

causing surface diffusion of the Pt (Reinhold et al., 2009). This drop in height

suggests that a reduced Pt shell has formed by the 10 min mark, rendering further

reduction ineffective. The Pt shells formed after 10 min plasma treatment are

visibly observable and can be pierced, resulting in the release of the liquid ink

underneath. Certain plasmas are able to reduce salts on their own, such as 900 W

H2-He plasma on PdCl5, which was found to have a maximum penetration depth of

25 μm achieved in 5 min (Koo et al., 2006). We have previously shown however that

low wattage, 30W in our case, plasma does not reduce H2PtCl6 without the

presence of EG, meaning it is not a direct plasma reduction mechanism. We have

also previously shown that while the surface may reduce, there may be unreduced

pockets/layers of ink depending on geometry (O’Connell et al., 2013). The longer

the plasma duration however, the more ruptures seem to occur. In this case, the

internal ink spreads on the newly hydrophilic substrate, and reacts with the plasma,

producing reflective films. In the case of thermal reduction on PDMS and plasma

treatment on Au-Mylar, there is no spreading as the contact angle stays constant

(Figure 4-10 D, F).

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Figure 4-10: Optical (A–C) and SEM (D-F) images of deposited features. (A) Unreduced ink
immediately after deposition; (B) unreduced ink immediately after 1 hr under vacuum exhibits no
spreading; (C) after 10 min plasma reduction. The presence of oxygen species create hydroxyl groups
on the surface of the PDMS, increasing its hydrophilicity and facilitating spreading of the ink
(Bhattacharya et al., 2005). (D) Thermally treated (200C) samples exhibit no spreading; (E) Plasma
treated samples on PDMS exhibit spread similar to C; (F) Plasma treated samples on Au-Mylar do not
exhibit spread, further supporting surface modification as the cause of spreading. Scale bars: 20 μm
(A-C), 100 μm (D-F).

The atmosphere of the plasma can be seen to have changed based on different

conditions (Figure 4-11). Vaporized EG creates light blue atmosphere (Figure 4-

11A). Normal air plasma is a deep purple (Figure 4-11B), while N2 is known to be

pink-orange in color (Foerch and Hunter, 1992). It can be seen that 5 pump cycling

the chamber before starting reduction reduces the amount of oxygen (Figure 4-

11C); this involved pumping the chamber down to 500 mTorr and back up to 1600

mTorr in cycles, before finally setting the chamber to rest at 1050 mTorr. After 30

min of plasma operation there is still yet another color change (Figure 4-11D),

suggesting there were still non N2 species present, such as oxygen, which may

modify the substrate. Attempts to measure the spectrum with a portable

Ryan P. Sullivan 152

spectrometer though the viewing window were unsuccessful. While having longer

pump cycles may reduce the oxygen species, which is both able to modify the

PDMS surface increasing the spreading and to convert reduced platinum into

platinum oxide, as will be explained in Sections 4.3.5 and 4.3.6, it is not worth the

tradeoff with EG lost due to evaporation.

Figure 4-11: Pictures of Pt-PDMS samples on glass slides and dishes in the plasma chamber during
treatment. A: EG atmosphere created by addition of mL droplets of EG; B: Air; C: The chamber was
pumped down to 500 mTorr and back up to 1600 mTorr 5 times before finally resting at 1050 mTorr;
D: post 30 min N2.

To act as a baseline for the reduced heights in Table 4-7, evaporation in normal

atmosphere was measured over a 22 min period at 1 min intervals (Figure 4-12). As

the contact angle 102° is greater than 90°, evaporation should keep the contact

angle constant, reducing the radius of the features. There was no noticeable

reduction in the area of the features over this time. Additionally histograms of the

heights (not shown) show no change when comparing the data from t = 0 and t = 22

min. Delays in reduction of deposited droplets after deposition impede the

reduction as will be shown in the following sections, which making establishing a

baseline evaporation in normal atmosphere important for determining the cause.

Ryan P. Sullivan 153

Figure 4-12: Areas of six features to measure evaporation of a 22 min interval after printing.
Measurements were made roughly every 1 min.

4.3.4 Penetration Depth of Dropcast Films

Reduction of bulk drop-cast samples was tested to analyze the penetration limit of

the plasma reduction seen in the larger deposited features. While in our previous

work we showed that bulk drop-cast reduced producing highly reflective films, X-

ray powder diffraction (XRD) showed small peaks belonging to unreduced material

among the main 111, 200, and 220 Pt peaks (O’Connell et al., 2013). While glass is

not a useful substrate for bionics, it does allow the material at the substrate-

deposit interface to be viewed directly. 0.5 uL of ink was dropcast on 1 x 0.5 cm

atmospheric plasma treated glass pieces; as can be seen in Figure 4-13 below, the

underside of deposited material did not fully reduce and remains black.

Ryan P. Sullivan 154

Figure 4-13: Reduced and unreduced sides of a 0.5 μL dropcast 10% Pt ink on atmospheric plasma
treated 1 x 0.5 cm glass pieces. The top side, in contact with plasma, is reduced and highly reflective.
The bottom side, as seen through the glass substrate, is brown/black and non-reflective.

When testing samples on non-plasma treated glass, the same volume was thicker

due to difference in contact angle. As can be seen in Figure 4-14A, N2 plasma

treatment produced the black material along most of the feature except for the

very edge. Successive N2 plasma treatments continued the reduction process until

the black material was all reduced leaving a highly reflective Pt sample (Figure 4-

14E).

Ryan P. Sullivan 155

Figure 4-14: Optical images of progression of reduction of 0.5 μL dropcast platinum ink on non-
plasma treated glass over repeated N2 plasma treatments. Duration listed is total duration. A) 20
min total plasma treatment produced mostly black material with metallic edges. B) 40 min total
plasma treatment caused islands of black material to recede, increasing the size metallic edges. C)
60 min total plasma treatment caused most of the material to reduce. D) 80 min total plasma
treatment completely reduced the top sample while leaving a small island of partially reduced
material in the center of the bottom material. E) H 2O washed sample using flash photography shows
the high reflectivity of the reduced Pt.

4.3.5 Conductivity

Conductivity measurements, performed using Conductive-AFM, were done on Au-

Mylar substrates instead of PDMS, as a conductive substrate is required to perform

the measurements. Time between deposition and reduction was found to strongly

affect conductivity. As seen in Figures 4-15 and 4-16, a delay in reduction after

fabrication leads to incomplete reduction of the ink. In the time it takes to conduct

optical profilometry measurements on deposited features (to analyze changes

before and after various plasma intensities and durations) changes to the deposited

features take place resulting in incomplete reduction. This is most likely either due

to evaporation of the reducing agent EG or a reaction with the atmosphere that

impedes the polyol process, though Figure 4-12 suggests evaporation is minimal.

Figure 4-15A is an AFM height image show highly heterogeneous and amorphous

structures with deposits of unreduced H2PtCl6 cubic salts (as confirmed by SEM-

EDS), pointed out by the blue arrows. Figure 4-15B is the corresponding

conductivity map for the same region. Yellow represents regions of high

conductivity, while dark red-black represent regions of low or no conductivity. The

blue arrows point out the unreduced H2PtCl6 cubic salts which show up as insulating

Ryan P. Sullivan 156

black regions in. Additionally, there is lower conductivity as a whole on the disk as

well as large height inhomogeneity as compared to the immediately reduced

sample (Figure 4-15D). The dark ring that can be seen around Figure 4-15D is most

likely ink residue remaining from when the droplet deforms upon initial impact,

before retracting to form the spherical cap (Rioboo et al., 2002). We have

previously showed that the low 30W N2 plasma will not reduce H2PtCl6 on its own

unlike higher wattage H2/He plasmas (Koo et al., 2006; O’Connell et al., 2013; Shim

et al., 2007; Wang et al., 2009). This residue from the receding droplet then forms

an insulating layer on the Au-Mylar substrate, causing a large contrast in the image.

Figure 4-15: Representative C-AFM images of reduced features on Au-Mylar. The black and white
images (A, C) are height maps of two different features; the yellow-red images (B, D) are current
maps of those same features. Samples with a delay of around 30 min between deposition and
reduction, 10min 30W plasma, (as shown in panels A and B) show low and heterogeneous
conductivity. Blue arrows point out unreduced Pt salt. Samples that were reduced immediately after

Ryan P. Sullivan 157

deposition, 10 min 30W plasma, (as seen in panels C and D) showed high homogenous conductivity
and no remaining Pt salt. Scale bars 10 μm.

Individual IV plots of the two features in Figure 4-15 can be seen in Figure 4-16. The

insulating region pointed out by the blue arrows corresponds to Figure 4-16A,

where pA capacitive charging can be seen (effectively horizontal IV response). The

general low conductivity of the delayed plasma reduced feature can be seen in

Figure 4-16B. The sample which was reduced immediately after printing however

shows high conductivity, almost vertical, similar to Au-Mylar (Figure 4-16C, D).

Figure 4-16: Representative C-AFM IV plots. A: capacitive charging of an unreduced insulating region
(depicted in Figure 4-15B by the blue arrows); B: reduced region that while conductive, has a
relatively high resistance (depicted in Figure 4-15B); C: highly conductive Au-Mylar substrate
(depicted in Figure 4-15C and D); D: highly conductive platinum sample reduced immediately after
deposition (depicted in Figure 4-15D).

Ryan P. Sullivan 158

Figure 4-17 shows average conductivity measurements on Au-Mylar substrates and

Pt features over a range of set points over 252 IV measurements. As can be seen in

Figure 4-17, the standard deviations are high compared to the actual

measurements. Previous measurements produced standard deviations an order of

magnitude higher than the average. It was found that high levels of noise were

caused by other equipment due to an unshielded mains; a second series of

experiments, as presented in Figure 4-17 and below, was undertaken after hours or

on weekends when there was no other usage on the same mains reducing the

noise. Taking all IVs for either the Au substrate or Pt feature gives an average

system resistance of 8.23 ± 3.31 and 8.54 ± 3.70 μΩ respectively. In order to

estimate conductivity as previously shown by (O’Hayre et al., 2004) (see Section

4.5.3), the bulk substrate resistance is subtracted from the measured resistance of

the substrate + feature, which gives an average resistance of 0.31 ± 3.51 μΩ for

each platinum feature. Assuming a cylindrical geometry as seen in the disk

morphology in Figure 4-15D, the final conductivity results in 78 nΩ*m, which is less

than bulk of 105 nΩ*m. The extremely high conductivity of solid metals pushes the

capabilities of the equipment, causing the op-amps to clip the current (Figure 4-16C

and D).

Ryan P. Sullivan 159

 16
14
12

Resistance (μΩ)
10
8
Au
6
Pt
4
2
0
0 0.1 0.2 0.3 0.4 0.5 0.6
Setpoint (V)

Figure 4-17: Plot of average total system resistance obtained from averages of C-AFM IV curves at
various set points. Error bars represent ±1 standard deviation.

A two sided ANOVA was run with metal (Au or Pt) and setpoint (0.1 - 0.5 V) as the

independent variables. The difference in resistance between the Au substrate and

the Au substrate + Pt features was found to meet our standard of statistical

significance (p <0.01), having an F value of 7.86 compared to F crit of 3.86, along

with a p value of 0.005.

4.3.6 Elemental Composition

Successful reduction should remove all chlorine in the form of HCl gas that

evaporates off (Bonet et al., 1999; Koo et al., 2005; Sun et al., 2010). In thermally

stimulated polyol processes, the main technique in the literature, the EG

byproducts are aldehydes, glycolic acid, oxalic acid, CO2 and carbonate, with

glycolaldehyde acting as the major reducing component (Fievet et al., 1989;

Skrabalak et al., 2008; Sun et al., 2010). Formation of glycolaldehyde is temperature

Ryan P. Sullivan 160

dependent; at temperatures above 140°C it will be the dominant reducing agent

while at temperatures below this EG is be the primary redundant (Fung et al.,

2010). As EG is converted into these smaller molecules, it should also evaporate off

due to higher vapor pressures. The elemental map seen in Figure 4-18, shows a high

concentration of Pt and Cl in the feature, indicating incomplete reduction. Oxygen is

lower in the location of the feature, and carbon is roughly the same. Oxygen is

found in both the reducing agent EG and in the substrate PDMS. Similar amounts of

carbon and the drop of oxygen suggest that there is negligible EG remaining in the

feature. This would explain the termination of reduction, as plasma of this power

has not been shown to reduce without EG except on single nanometer scales

(O’Connell et al., 2013).

Table 4-8 below shows the percentage masses of each element over multiple

features along with their elemental ratio with respect to Pt. The boxes in red

highlight the ratio of Cl to Pt. Unreduced, this ratio is 6:1. Thermal reduction fares

better than plasma at both 5 W and 30 W, and microwave (1 kW), but still shows a

1:1 ratio of Pt:Cl. There is improvement on Au-Mylar substrates, most likely due to

the different contact angle, which results in reduced feature thickness for the same

volume of deposited material, thereby reducing the effects of any penetration limit.

When comparing Au-Mylar substrate samples, there is a large difference in the final

ratio when the reduction was performed immediately after deposition, without

optical profilometry measurements. In this case the ratio dropped to 0.14:1 in

certain regions, supporting the conductivity data in Figures 4-15 and 4-16,

demonstrating that increased time between deposition and reduction strongly

Ryan P. Sullivan 161

impedes reduction of the ink. The 5 W plasma settings were used in an attempt to

more evenly reduce salts in the ink without forming a shell. The results here

indicate this did not fix the issue.

Figure 4-18: Lowvac SEM EDS Maps of a Pt feature on PDMS The top left image is the SEM image.
The other five images are elemental maps of (clockwise) Pt, Cl, C, O, and Si. High amounts of Cl
indicate unreduced material. Equal amounts of C and lower amounts of O relative to the substrate
indicated absence of EG. Together, this suggests that evaporation of reducing agent halted Pt
reduction. Scale bars: 10 μm.

Table 4-8: Average EDS values of 7 features per parameter, taken using Low-Vac SEM of 10 pL
deposited 10% Pt weight inks on PDMS and Au-Mylar. Cl ratios are highlighted in red. Complete
reduction should have Cl ratio of 0:1 while incomplete reduction should have a Cl ratio of 6:1.

Ryan P. Sullivan 162

 Reduction Method - Substrate C O Cl Pt Au Si
% Mass 13.23 14.90 15.08 32.39 24.40
Plasma (5 W 10 min) - PDMS
Ratio:Pt 6.64 5.61 2.56 1.00 5.25
% Mass 12.87 14.51 19.95 30.04 22.62
Plasma (5 W 20 min) - PDMS
Ratio:Pt 6.96 5.88 3.65 1.00 5.24
% Mass 11.53 16.80 20.14 30.81 20.72
Microwave (1 kW 3min) - PDMS
Ratio:Pt 6.08 6.65 3.59 1.00 4.68
% Mass 8.08 10.62 9.58 54.30 17.41
Thermal (200C 90 min) - PDMS
Ratio:Pt 2.42 2.38 0.97 1.00 2.23
% Mass 28.54 9.02 7.71 29.95 26.43
Plasma (30 W 10 min) - Au
Ratio:Pt 15.49 3.67 1.42 1.00 0.87
% Mass 30.79 11.57 0.59 22.83 34.22
Plasma (30 W 10min) - Au Thin
Ratio:Pt 21.92 6.18 0.14 1.00 1.48

4.3.7 Ink-Inkjet Head Interactions

Allowing the filled ink cartridge sit for an hour or two before printing was found to

increase the jetting stability (FUJIFILM Dimatix, Inc., 2010), allowing for increases in

printing frequency up to 10 kHz. In order reduce the setup time for successive

printing sessions, filled cartridges were let sit vertically outside the printer, thereby

allowing immediate printing at faster firing frequencies. However it was found that

the ink can damage the hydrophobic coating on the inkjet head, causing ink to build

up at the edges, disrupting further jetting, as seen in Figure 4-19 below. At the end

of every print session, the ink cartridges were manually purged with pure EG until

the orange color of the H2PtCl6 salt was no longer visible. At the beginning of the

subsequent print sessions, the cartridges were again purged, but with platinum ink,

until the ink output was consistently orange. In cases where there was a long

interim period between using a given cartridge, e.g. at least a week, the cartridges

were purged with IPA. The high volatility of IPA allowed the cartridges to

completely dry out so that they could be stored long term. This manual purging is

tactically demanding. If the cartridges were purged too quickly by applying too

Ryan P. Sullivan 163

much pressure, the paths to the inkjet array ruptured causing ink to be expelled

from the side of the inkjet plate. At the end of purging, the applied pressure needed

be lowered slowly; if it was lowered too fast, it would create back pressure, and air

would be pulled into the openings of the inkjets, destabilizing or completely

inhibiting proper droplet formation.

Figure 4-19: Optical image of ink jet head. Damage around the edge of the square head can be seen
where the ink has caused the surface to break away. When this occurs, droplets formed during
jetting can be pulled into this now hydrophilic sections, altering jetting angle, causing split droplets,
or even stopping jetting altogether. Scale bar: 20 μm.

4.4 MEDICAL READINESS

Ryan P. Sullivan 164

There are two main potential hurdles that polyol reduced features will need to

overcome to be used in medical implants. The first is to ascertain that there are no

toxic byproducts left from the polyol reduction, while the second is ensuringstrong

homogenous adhesion to the substrate. Delamination of thin film electrodes has

been shown to be a large area of concern (Ordonez, et al., 2015) During constant

cycling, the surface of electrodes may become oxidized and react with organic

molecules. This change in surface chemistry may penetrate the electrode on the

nanometer scale, changing the lattice constants of the film. As the lattice constants

are changed, this induces stress in the film. In thin films, this changed lattice may

affect come to dominate the internal stresses, whereas with traditional electrodes

these stresses may be viewed as negligible (Lacombe et al., 2006). One of the

reasons that the PDMS used in this chapter was cured at a lower temperature was

to preserve some of the reductive capacity of its precursors. The PDMS precursor

formulation uses a platinum based reduction mechanism; it has been shown that

metal salts, particularly platinum, gold, and silver, that are dispersed in this PDMS

precursor are reduced as the PDMS crosslinks producing fully reduced

nanoparticles. By taking a similar approach, the reduced platinum on at the PDMS

interface should bond using coordination chemistry, where each platinum atom

may create 2 bonds each to the PDMS below. This should serve to anchor the film

and reduce the effect of lattice changes as been shown previously (Green et al.,

2014) as the bulk substrate absorbs some of the stress.

A second large issue to delamination is induced stress from the delta between high

deposition temperatures and the final resting temperature of the film and substrate

Ryan P. Sullivan 165

(Ordonez, et al., 2015). As polyol reduction does not have elevated temperatures

during metal film formation, the differences in the Coefficient of Thermal between

film and substrate becomes neglibable.

Our experiments showed that while unreduced material is easily dislodged and

removed during contact AFM or isopropanyl sonication, reduced features are not

(Figure 2-13). In fact, the reduced features adhered sufficiently strongly, that they

were causing the platinum coated tips to break, requiring conductive diamond tips

to be used for consistent current mapping. Future in depth studies will need to be

performed to fully quantify delamination potential, but it is of more importance to

test the scalability of the technique to line and electrode formation before large

films can be tested.

In terms of byproduct toxicities, the main molecules of concern are

unreduced/partially reduced salt, HCl, EG, EG products such as actyl groups. Platinic

acid and its intermediates are corrosive my nature. HCl LD50 238 mg/kg, a

byproduct of platinic acid decomposition, has a high solubility in water and will

rapidly dissociate for form hydronium ions, which can cause cell death

(AquaPhoenix Scientific 2015). EG is neurotoxic due to how it is metabolized and is

classified as Toxicity Class III, LD50 4700 mg/kg (ScienceLab.com 2013). In particular

it is transformed into glycolaldehdye, and further oxidized to glycolic acid, which

may lead to edema (Brent et al.,2001, Maier et al., 1983). The crystallization of

PDMS from plasma treatment may create SiO2, which has a LD50 (oral) of 3160

mg/kg compared to PDMS 40,000 mg/kg. Based on the elemental data in Table 4-8,

Ryan P. Sullivan 166

there are most likely byproducts of some form from both chlorides and EG. Future

work to achieve more complete reduction, such as using higher powered plasma or

direct thermal reduction with lasers, may more completely remove these traces.

4.5 CONCLUSIONS

In this chapter we have successfully extended the use of polyol inks from DPN to

Inkjet printing. We have developed and optimized jetting waveforms to produce

highly ordered and reproducible platinum patterns 22.89 ± 4.79 μm diameter in

resolution at rates of 48,000 features per second with 16 jets. While the stable 3

kHz deposition rate that we were able to achieve is considerably higher than that of

DPN, it is only 20% of the maximum firing rate of the particular jets used on the

inkjet printer (FUJIFILM Dimatix, Inc., 2010). While we were able to achieve firing

rates of up to 10 kHz (160,000 features per second), this was not a reproducibly

stable firing rate between multiple jets. We were also unable to determine exactly

at which firing rate the instability sharply increases for the chosen waveform. While

continued work sampling the different frequencies may elucidate this limit, it may

be more productive to alter components of the waveform to reduce the time until

the pressure waves have dissipated and returned to baseline.

We have shown that printing lifetime is a large constraint on the successful

reduction of polyol inks to metal. We have demonstrated that the high conductivity

of platinum features is on the same order as bulk platinum, 78 nΩ*m vs 105 nΩ*m

respectively, based on the assumption of a simple cylinder. However, due to the

large noise and deviation of measurements, and the fact that nanoscale resistivity is

Ryan P. Sullivan 167

often lower than resistivity of larger scales, further work needs to be carried out to

determine conductivity of lines, such as a Greek Cross structure for 4-point probe

tests. Additionally, there is a limit to the thickness of the printed material that

plasma will reduce. While Pt polyol inks can be highly controlled for stable jetting

needed for large scale fabrication, further reduction parameters needs to be

examined to overcome penetration depth of plasma reduction. Alternative

reduction approaches such as localized thermal reduction via lasers, as used in

other ink jet setups, may overcome these limits, as well as reduce the total

fabrication time, as currently the 10 min plasma treatment is considerably longer

than the seconds needed to deposit patterns. As the high contact angle of the ink

on PDMS leads to formation of spherical caps, modification of the surface will be

vital to eventually form conductive lines. Such an approach will be the focus of the

next chapter.

Ryan P. Sullivan 168

4.6 APPENDIX

4.6.1 Vapor Pressure of Ink

The following is the derivation for vapor pressure of platinum inks depending on

loading. We start with Raoults Law:

𝑃𝑣𝑎𝑝 = ∑ 𝑝𝑖 𝜒𝑖 (1)
𝑖

where P is the vapor pressure, 𝜒𝑖 is the mole fraction, and 𝑝𝑖 is the partial pressure.

In the case of two part polyol inks:

𝑃𝑣𝑎𝑝 = 𝑝𝐸𝐺 𝜒𝐸𝐺 + 𝑝𝑠𝑎𝑙𝑡 𝜒𝑠𝑎𝑙𝑡 (2)

As the salt is a solid, its partial pressure effectively goes to zero, therefore:

𝑚𝐸𝐺 (3)
𝑃𝑣𝑎𝑝 = (7.5𝑃𝑎)
𝑚𝐸𝐺 + .318𝑚𝑃𝑡

𝑚𝑃𝑡
+ (4.47 ∗ 10−3 𝑃𝑎)
𝑚𝑃𝑡 + 3.14𝑚𝐸𝐺

1 − 2.101𝑚𝑃𝑡
= (7.5𝑃𝑎)
1 − 2.101𝑚𝑃𝑡 + .318𝑚𝑃𝑡

𝑚𝑃𝑡
+ (4.47 ∗ 10−3 𝑃𝑎)
𝑚𝑃𝑡 + 3.14(1 − 2.101𝑚𝑃𝑡 )

1 − 2.101𝑚𝑃𝑡 1 𝑚𝑃𝑡
= (7.5𝑃𝑎) + (4.47 ∗ 10−3 𝑃𝑎)
1 − 1.783𝑚𝑃𝑡 3.14 1 − ⁡ 1.783𝑚𝑃𝑡

𝑚𝑃𝑡 1
= [(7.5)(1 − 2.101𝑚𝑃𝑡 ) + (4.47 ∗ 10−3 ) ]
3.14 1 − ⁡ 1.783𝑚𝑃𝑡

1
= [7.5 − 15.758𝑚𝑃𝑡 + 1.422 ∗ 10−3 𝑚𝑃𝑡 ]
1 − ⁡ 1.783𝑚𝑃𝑡

Ryan P. Sullivan 169

 7.5 − 15.757𝑃𝑡% (4)
𝑃𝑣𝑎𝑝 = 𝑃𝑎
1 − ⁡1.783𝑃𝑡%

4.6.2 Error

1000.00%

800.00%
Area (% of actual)

600.00%

400.00%

200.00%

0.00%

-200.00%
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
Diameter (pixels)

Figure 4-S1: Graph of error in area (light blue shading) as a percentage of the true area (dark blue
line) as a result of single a pixel difference on thresholded edges. In the case of the spherical
geometry of the droplets cross-sections, the diameter changes ±2 as they have two edges.

4.6.3 Conductivity

Assuming a circular contact, the spreading resistance RSR is

𝜌
𝑅𝑆𝑅 =
4𝑟

where r is the radius, and 𝜌 is the resistivity of the sample.

Faradaic resistance and double layer capacitance scale with the electrode.

electrolyte contact area:

Ryan P. Sullivan 170

 𝑅𝑓0
𝑅𝑓 = 𝐴 𝐶𝑑 = 𝐶𝑑0 𝐴𝑐𝑜𝑛𝑡𝑎𝑐𝑡
𝑟𝑥𝑛

where 𝑅𝑓 is Faradaic resistance (Ω), 𝑅𝑓0 is the specific Faradaic resistance (Ω cm2),

Arxn is the available reaction surface area (cm2), Cd is the double layer capacitance

(F), 𝐶𝑑0 is the specific interfacial capacitance (F/cm2), and Acontract is the true

electrode/electrolyte contact area.

Contact Area from Contact force

Assume the hardness (H) of a material is constant (O’Hayre et al., 2004):

𝑃
𝐻=
𝐴

where H is hardness (N/m2), P is the applied force (N), and A is surface area of

contact (m2). H is constant for noncrystalline materials and geometrically similar

probes (O’Hayre et al., 2004). H is constant for crystalline materials with small

indentation size effect ISE (O’Hayre et al., 2004). H may be based either on project

area of contact (e.g., Knoop and Berkovich Hardness) or the surface area of contact

(e.g., Brinell, Vickers Hardness) (O’Hayre et al., 2004). H is related to mean

pressure:

P
Pm =
Aproj

Where Pm is the mean pressure (N/m2), P is the applied force (N), and A is the

projected area of contact (m2).

Ryan P. Sullivan 171

 CHAPTER FIVE: 1 PICOLITRE INKJETTING OF
PLATINUM INKS

5.1 INTRODUCTION

Neuroelectrodes contain different types of materials that are difficult to

manufacture together. While current FDA approved commercial implants are

mostly limited to platinum electrodes, or alloys thereof, with silicone rubber

insulators, there are many additional materials and techniques on the horizon.

These range from conducting polymer coatings for the electrode surfaces and

patterned biologics to direct regrowth, to hydrogel encased cell populations and

controlled release factors for targeted population regeneration and growth

(Thompson et al., 2016). Inkjet is a technique that has been demonstrated to be

able to print all of these various materials (Fribourg-Blanc et al., 2013; Layani et al.,

2012; Reinhold et al., 2009). As such, it holds promise for being an all-in-one

additive fabrication approach for neural implants tailored for the patient. This is

important, as the scala tympani for instance, where part of cochlear implants are

inserted, differs largely in both size and shape from patient to patient (Rebscher et

al., 2007, 2008). As such, it is important to help complete the picture by optimizing

platinum line printing for the electrode components of neural implants.

Ryan P. Sullivan 172

Ink-substrate interactions are critical for successful inkjet printing of lines. Control

of the surface energy and wetting of a substrate is important in order to prevent

both bleeding at high contact angles, and blebbing or excessive spreading at low

contact angles (Moon et al., 2014). The roughness and porosity of substrates also

play strong roles in pinning the edges of the ink (Alava and Dubé, 2012).

Additionally the adhesion between the deposited material and the substrate is

important to prevent delamination of the deposited material from the substrate in

the long term. PDMS, a silicone which is often used in medical implants such as the

Cochlear and Argus II for its chemical inertness and long term biocompatibility, is

unfortunately very hydrophobic, smooth, non-porous, and has poor adhesion to

metals, which make printing on a PDMS substrate extremely difficult (Wu et al.,

2014). As seen in Chapters 2 and 4, these properties are responsible for the

production of near ideal spherical caps regardless of ejection velocity and

coalescence into larger spheres.

Plasma treatment is often used on substrates to encourage proper line formation

due to its ability to increase wettability and surface energy (Fleischman et al.,

2012). The effects of various plasma species and parameters on PDMS have been

the subject of much study over the last two decades (Bacharouche et al., 2013;

Barbier et al., 2006; Bhattacharya et al., 2005; Bodas et al., 2008; Chu et al., 2002;

Fuard et al., 2008; Hillborg and Gedde, 1999; Hino et al., 2008; Kim and Jeong,

2011; Meincken et al., 2005; Morent et al., 2007; Sohbatzadeh et al., 2013; Williams

et al., 2004). Surface modification with adhesion promoters such as polydopamine

(PDA) have also been used to modify PDMS and other substrates (Chuah et al.,

Ryan P. Sullivan 173

2015; Jun et al., 2014; Leung et al., 2015; Lin et al., 2013; Meng et al., 2016; Zhang

et al., 2013). During the period from January to May of 2016 (the time of this

writing), 207 peer reviewed papers were published on PDA. The use of PDA for

biomaterials as well as its ability to reduce metal salts doubly improves upon its

uses explored in Chapter 2 where it facilitated metal line printing (Liang et al., 2011;

Meng et al., 2016).

In this Chapter, we therefore extend upon the surface modification treatments

used in Chapter 2, for use with Inkjet printing. Additionally, we have continued our

work on inkjet waveform design from Chapter 4, to reduce the size of deposited

features, moving from 10 pL to 1 pL ink jets.

5.2 EXPERIMENTAL DETAILS

5.2.1 Ink Creation, In-Flight Jetting, and PDMS Substrate Creation

The creation of the ink, the in-flight jetting and PDMS substrate creation used for

this Chapter were performed according to the protocols described in detail in

Chapter 4 sections 4.2.1 to 4.2.3, with the exception being that 1 pL jets were used

in place of 10 pL jets.

5.2.2 Inkjetting of Grids

1.5 mL of the prepared metal ink was loaded into Diamatix cartridges and grids of 5

x 4 dots were printed on PDMS substrates using the 1 pL jets. Total pulse duration,

applied voltage, and firing frequency of the waveform were altered over ranges of

8-12 μs, 25-40 V, and 1-15 kHz respectively. The printed 5 x 4 grids were used as the

Ryan P. Sullivan 174

unit for a larger 5 x 5 grid. Each unit grid was then imaged at x27.3 magnification on

a Wyko NT9100 Optical Profilometer (Veeco, USA). The resulting images were then

processed using the ImageJ plugin described in Chapter 3. Results were then

compared using Mann-Whitney U tests in SPSS to ascertain statistical significance,

with significance set to p < 0.01.

Grids of 6 x 5 dots were also printed on glass substrates to compare impact

characteristics on harder, less hydrophobic substrates.

5.2.3 PDMS Plasma Modification

To facilitate line printing, PDMS samples were plasma treated (PDC-002, Harrick,

USA) using atmospheric plasma at 30 W for 10, 30, and 120 s to modify the surface

with hydroxyl groups, increasing the hydrophilicity of the PDMS substrate.

5.2.4 PDMS Modification with Polydopamine

To facilitate line printing, PDMS samples were plasma treated (PDC-002, Harrick,

USA) for 10 min using atmospheric plasma at 30 W and 1100 mTorr to add hydroxyl

groups onto which the dopamine could react and anchor. The PDMS samples were

then placed in a solution of dopamine in Tris Buffer pH 8.5 (1 mg/mL) and were

gently agitated on an orbital shaker for either 15 or 60 min. The PDMS samples

were subsequently removed, rinsed under deionized water for 10 s, and dried

under N2 flow.

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5.2.5 Inkjetting of Lines

Lines of platinum ink were printed on untreated PDMS, plasma treated PDMS, and

dopamine treated PDMS. The printed samples were then reduced with a N2 plasma

treatment for 10 min at 30 W immediately after (within 10 s) the completion of the

ink deposition. Resulting features were characterized using the Optical Profilometer

at various magnifications. The intensity channels were extracted and analyzed with

the Grid Analyzer and Analyze Stripes ImageJ plugins.

5.2.6 Multimeter Conductivity

The resistance of 3 cm long PDMS-PDA-Pt samples were characterized using a

multimeter by placing leads at each end of the line. Leads were moved along the

lines in order to account for contact resistance.

5.2.7 Scanning Electrochemical Microscopy

PDMS-PDA-Pt samples were characterized using a SECM (CHI SECM) using raster

scanning with a 10 μm diameter Pt microelectrode in 0.1 M KCl with 1 mM

ferrocene methanol as the redox mediator.

5.2.8 Conductive AFM

PDMS-PDA-Pt samples were characterized with C-AFM (MFP-3D Classic, Asylum

Research, USA) in Orca mode with a conductive cantilever tip. Samples were biased

at the ends of deposited platinum lines and measurements taken with the AFM tip

in contact with parts of the line. Topographical and current images were obtained,

along with point IV plots.

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5.2.9 Sonication Adhesion Test

PDMS-PDA-Pt samples that had undergone 10 min N2 plasma treatment lines were

immersed in water and sonicated for 10 min to test the adhesion of printed

features to their substrates.

5.3 RESULTS AND DISCUSSION

5.3.1 In Flight Jetting Analysis

As in Chapter 4, droplet formation and flight immediately adjacent to the jets was

observed using a μsec strobe to obtain still images of droplets.

5.3.1.1 Drop Volume

The issues of low contrast and high background noise of the previous 10 pL images

(Figure 4-3) were found to be exacerbated for the 1 pL images (Table 5-1). This was

likely due to the same optics being used to capture the images, despite the much

smaller volume of the droplets being jetted. The diameters of the slices range from

~1-9 pixels for the 1 pL droplets, which is equivalent to 2.6-23.7 μm. There was no

significant trend with either pulse duration or applied voltage, unlike that found for

10pL results in Table 4-2. For many of the parameters, it was extremely difficult to

capture images. At many parameters jetting was only transiently stable, become

erratic after only a short time. Blank cells in Table 5-1 or those with no recorded

standard deviation are such because either no samples were able to be obtained, or

only one sample was able to be obtained for the respective parameters.

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Table 5-1: Average volume data (pL) obtained by summating circular slices perpendicular to the travel vector of droplets in flight with 3 individual ink jets. Blank cells
indicate no data could be obtained for given parameters in any of the 3 jets. Lack of standard deviations indicates that only a single jet produced quantifiable data.

Pulse Width 20 V 21 V 22 V 23V 24V 25V 26V 27V 28V 29V 30V
8 μs 4.8 6.4 6.0 5.0 ± 2.9 5.1 ± 0.4 7.1 ± 1.1 4.5 ± 0.5 8.8 ± 6.1
9 μs 2.4 ± 3.4 8.0 ± 5.0 10.2 ± 9.6 6.2 ± 1.1 5.1 ± 0.7 5.4 ± 2.1 5.4 ± 1.4 5.6 ± 2.3 5.7 ± 1.9 5.7 ± 2.0 5.1
10 μs 7.9 ± 3.6 5.6 ± 0.2 5.8 ± 1.2 5.7 ± 0.6 7.6 ± 2.6 4.9 ± 0.6 4.9 ± 0.5 4.2 ± 0.5 5.6 ± 1.7 4.4 ± 0.3 5.1 ± 0.8
11 μs 4.7 5.6 6.6 5.4 4.6 ± 0.3 5.7 ± 2.0 5.5 ± 2.1 6.8 ± 3.7 5.0 ± 0.4 5.0 ± 0.8 4.2 ± 0.6
12 μs 4.4 5.2 4.9 5.3 5.5 3.5 7.9 4.4 4.6 4.3 3.0

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5.3.1.2 Drop Velocity

The velocity profiles for the 1 pL droplets were very similar to those of the 10 pL

droplets. There was a clear trend with increasing voltage, but no such trend in

terms of pulse duration (Figure 5-1). 11 μs was an outlier, having roughly half the

velocity of the other pulse durations at 20 V. At 26 V and above, it became difficult

to measure many of the droplets due to tail formation and positioning of the optics,

though summation of circular slices (Figure 4-4) still allowed for the calculation of

droplet volume. This means that 1 pL jetting would be expected to produce similar

impact phenomena to 10 pL, which may become important for multiple layer

printing in future experiments.

Figure 5-1: Velocities of 1 pL droplets of 10% Pt weight ink as a function of the applied voltage (V)
and pulse length (μs). There is a clear positive relationship for voltage. With the exception of the 11
μs outlier, all pulse durations tend to group together.

5.3.1.3 Drop Coalescence

In terms of distance until coalescence, 1 pL droplets express the same trends as

found with 10 pL droplets. Higher applied voltages cause droplets to travel further

Ryan P. Sullivan 179

before they coalesce into a sphere as seen in Figure 5-2. Increased duration also

decreases the distance until drops coalesce. As with the 10 pL experiments, the ink

jet head was set above the range of drop coalescence so there were no issues

impacting before droplets had completely coalesced.

Figure 5-2: Distance until coalescence of 1 pL droplets of 10% Pt weight ink as a function of the
applied voltage (V) and pulse length (μs). Excluding the 11 μs pulse length, which is an outlier in this
parameter as well as velocity (Figure 5-1), there is a negative relationship between both pulse
duration and velocity with the distance traveled until the droplet forms a sphere.

5.3.2 Inkjetted pattern analysis

While a large space was explored in flight, the resulting patterns were highly

disordered. All combinations at 10 and 15 kHz resulted in spraying of the ink. While

combinations at 3 kHz produced normal droplets instead of spray, the position of

the droplets were highly erratic to the point that values for theta, X, and Y could not

be calculated for most of the parameters. 10 μs - 30 V, which was the most stable

set of parameters for 10 pL in the previous chapter, was the only parameter

combination to reliably produce measurable grids on the scale of 1000 s of features

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per print (Figure 5-3D). While voltages higher than 26V resulted in difficulties

measuring the inflight properties of droplets due to leaving the viewing area, the

pattern analysis does not suffer from this deficiency, allowing analysis up to the

maximum voltage of 40V.

As seen in Figure 5-3A and C, higher magnification images were taken of the 1pL

grids due to their smaller size. Additionally grids were set at 5 x 3 instead of 6 x 5 to

better fit inside the field of view to stay robust in cases where grids had high

disorder, which would stretch the bounds of the grids.

Figure 5-3: A and C are pattern schematics for the calibration grids shown in optical images B and D
respectively. A and B represent grids jetted using 10 pL jets while C and D represent grids jetted
using 1 pL jets. Both jets used the same parameters of 10 μs 30 V 3 kHz on PDMS substrates. Scale
bars: 100 μm.

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Compared to features printed with 10 pL jets in the study in Chapter 4 (Figure 5-

3B), features printed using 1 pL jets in the present study (Figure 5-3D) were shown

to have a 66.3% decrease in diameter, 22.89 ± 4.79 μm vs 10.08 ± 2.21 μm (Table 5-

2). As discussed in the previous two chapters, the stability of jetting is measured

through the parameter theta, which is the angle formed by a given droplet and its

two neighbors. An ideal grid has all right angles; therefore stability is said to

decrease as the average theta value deviates from 90°. The smaller size of features

produced using 1 pL jets was found to come at a slight cost to stability, with theta

dropping roughly 1.6° on average, p-value = 3.49x10-14, (an equivalent 289%

increase in deviation from 90°) and tripling the standard deviation (from 89.1° ±

1.0° to 87.4° ± 3.0°). While the deviations of the X and Y distances (discussed in

Chapter 3, section 3.3) are proportionately larger for 1 pL jetting than they are for

10 pL jetting (0.1 ± 3.0% vs 10.0 ± 6.3% for the X distance and 1.3 ± 3.5% vs 5.7 ±

13.4% for the Y distance), it is important to note that part of the deviation in these

values is attributed to the mechanical accuracy of the actual cartridge. While the

average recorded difference in distance is lower for the 10 pL cartridge (0.1 vs 1.97

μm and 1 vs 1.14 μm), the spread, i.e. standard deviation, is actually higher (± 2.2 vs

± 1.25 μm for X distances and ± 2.8 vs ± 2.68 μm for Y distances). Relative to the

magnitude of the individual pitches though, 1 pL has a larger effect. Looking at

Figure 5-3, the vertical deviation, seen as Y values in Table 5-2, is more apparent in

the 1 pL grid (Figure 5-3D) than the 10 pL grid (Figure 5-3B) even though they are on

average roughly equal, being 1.14 μm and 1.0 μm off from the set pitches

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respectively. In terms of percentage, the difference is 5.7% of the pitch for 1 pL but

only 1.3% of the pitch for 10 pL.

Table 5-2: Geometric values of printed drops obtained from the ImageJ plugin. Both size jets use the
same waveform parameters. Other parameter combinations are not shown as they were too
unstable to adequately quantitate order values. 1 pL and 10 pL interdot distances were 20 μm and
80 μm respectively. N=3240

Volume Size Theta X Y

10.08 ± 2.21 μm
1 pL 79.86 ± 3.82 μm2 87.4° ± 3.0° 21.97 ± 1.25 μm 21.14 ± 2.68 μm
0.23 ± 0.00 pL
22.89 ± 4.79 μm
10 pL 411.41 ± 17.98 μm2 89.1° ± 1.0° 79.9 ± 2.2 μm 79.0 ± 2.8 μm
2.66 ± 0.02 pL

Grids were printed on a cleaned glass substrate to act as a comparison for harder

and less hydrophobic substrates. Since PDMS is relatively soft and highly

hydrophobic, any issues caused by a high jetting velocity on impacts were masked,

as the drops receded to near perfect spherical caps with negligible pinning. By

analyzing glass samples, we can better understand how the ink and waveform

perform in the face of pinning. We observed a decrease in the circularity of printed

features on glass compared to those printed on PDMS (Table 5-3), though it was

within the standard deviation. However there was a large difference in terms of the

observed roundness, with features printed on glass showing decreased roundness

compared to features printed on PDMS, with an increase in the standard deviation

by a factor of 20 (Table 5-3). This indicates that while features on PDMS and glass

are equally compact, the perimeter heterogeneity is significantly increased on glass

substrates. This can be attributed to an increased prevalence of pinning on the glass

surface compared to PDMS. When comparing 10 pL and 1 pL drops on PDMS, the

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lower circularity of 10 pL droplets may be partially explained by induced anisotropy

of the images from the interference pattern when they were taken, as can

minimally be seen in Figure 5-3 B and D.

Table 5-3: Average circularity and roundness values for features. Circularity values are roughly the
same for PDMS and glass, and are larger for 1 pL compared to 10 pL on the same substrate. There is
a much larger difference in roundness however between the PDMS samples and the glass samples.
This is attributed because roundness is more a measure of perimeter uniformity compared to
compactness of circularity. The glass substrate has more pinning on the edges after impact, which
shows up as a decrease in the roundness value. N = 3240

Circularity Roundness
10 pL PDMS 0.909 ± 0.16 0.962 ± 0.004
1 pL PDMS 0.941 ± 0.012 0.952 ± 0.005
10 pL Glass 0.894 ± 0.44 0.853 ± 0.097

5.3.3 Substrate Modification

Use of the 10 pL parameters developed in Chapter 4 were used over the 1 pL

parameters developed in this chapter due to the aforementioned stability. 10 μs

30V waveforms were used for all line printing discussed in this section. As stable

jetting had been achieved, the next step of interest is how multiple droplets

interact when they overlap. This is dictated by the droplet-substrate interaction,

mainly hydrophilicity in our case, as well as the deposited patter, with parameters

of drop overlap, the thickness of lines in terms of drops, and the number of layers

of droplets deposited.

Attempting line printing on PDMS was unsuccessful due to the extreme

hydrophobic nature of the PDMS substrate. Even at scales of half a centimeter in

width, the ink would bead up into discrete spherical caps as seen in Figure 5-4B.

Plasma treatment is a commonly used technique used to modify the surface

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hydrophilicity of PDMS (Zhou et al., 2010). PDMS surfaces were modified for 10, 30,

and 120 s under atmospheric plasma. 120 s of treatment caused the ink to spread

multiple millimeters and rapidly evaporate due to the increase in surface area.

When shorter treatment times were used, the PDMS rapidly regained its

hydrophobicity, causing the ink to initially spread and then subsequently bead as

seen in Figure 5-4D. This is a known issue with PDMS modification as the newly

hydrophilic strands will reorient away from the surface and towards the bulk (Zhou

et al., 2010). Low curing temperatures were used because it has been previously

shown that PDMS oligomers can reduce metal salts (Goyal et al., 2009), which

would increase the adhesion of the resulting metal features. However, the low

curing temperature would increase the mobility of the strands, exacerbating the

issue of hydrophilic strands migrating towards the bulk.

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Figure 5-4: A and C are inkjet pattern schematics of printed “lines” on PDMS in B and D respectively.
Circles in A and C represent individual droplet placements, with each circle being 20 μm in diameter.
Deposited ink in both B and D is unreduced to examine the substrate-ink interactions. B is an
untreated PDMS substrate while D is a PMDS substrate treated with atmospheric plasma for 30 s
before deposition. In the case of C, the true size of individual features is unknown due to spreading.
Any lines printed on cured PDMS bead up to individual drops at all sizes attempted even at the mm
scale. In the case of 30 s atmospheric plasma treatment, the surface becomes highly hydrophilic
facilitating spreading of the ink. However, it is unstable and as it regains hydrophobiciity the spread
ink starts to bead up again as seen in B. Scale bars (A and B): 200 μm; (C and D): 500 μm.

Polydopamine (PDA) modification has been shown to reduce metal salts, including

H2PtCl6 specifially (Liu et al., 2013). PDA has also been commonly used as an

adhesion layer (Zangmeister et al., 2013). To overcome the limits of ink beading due

to the high contact angle on PDMS, PDA thin films were synthesized for a duration

of 15 min, as in Chapter 2. It was found that while the polydopamine film strongly

increased adhesion and decreased the contact angle, line printing resulted in

excessive blebbing. A treatment duration of 60min however produced considerably

improved line printing as seen in Figure 5-5. This may be due to changes in the

Ryan P. Sullivan 186

resulting film as the polymer chains lengthen, or due to increasing roughness with

duration (Zangmeister et al., 2013). In the literature some protocols call for multiple

hours, even up to 24 hr of synthesis time (Kang et al., 2011; Yang and Zhao, 2011),

so both durations used in this study fall on the shorter side of treatment periods.

Figure 5-5: Photograph of reduced printed platinum features on PDMS-PDA substrates with ruler for
scale. PDA synthesis durations were 15 min for samples A and B, and 60 min for samples C and D.
Sample A had 5-pump cycling before plasma reduction while sample B had a single pump cycle
before reduction. While the extra pump cycling would increase the percentage of N 2 species and
reduce all other species presents, the extra vacuum time inhibited reduction by allowing extra
evaporation of the solvent. Sample A delaminated immediately upon contact with water droplets
while sample B survived 10 min sonication. Increasing from 15 min PDA synthesis duration (B) to 60
min (for both samples C and D) reduced blebbing and allowed straighter line formation. Sample C
had lines with thickness set to 500 μm and was printed at 10 kHz while sample D had lines with

Ryan P. Sullivan 187

thickness set to, left to right, 100, 200, 100, and 100 μm (the fourth line is printed along the edge of
the sample and may be difficult to see) and was printed at 3 kHz.

The reduced Pt lines were reflective to the point that the image contrast caused

PDMS substrates to appear black when using the optical profilometer. This can be

highlighted by comparing the reduced features in Figure 5-6 B and D below, vs

unreduced features in Figure 5-4 B and D above. The amount of drop overlap, line

thickness, and therefore height of the line strongly influence the reduction. The

dark crack-like features throughout Figure 5-6D and at the edges on the right side

of Figure 5-6B are raised features, according to optical profilometry and AFM height

maps (not shown). We attribute this to the thickness of deposited lines limiting the

plasma facilitated reduction.

Figure 5-6: A and C are schematics of the inkjet patterns used to print the lines shown in B and D
respectively. Circles in A and C represent individual droplet placements, with each circle being 125
μm in diameter. A has width of 2 droplets with a 50 μm inter-drop spacing, producing a total width

Ryan P. Sullivan 188

of 175 μm. C has a width of 5 droplets with a 40 μm inter-drop spacing, producing a total width of
325 μm. Thicker lines, produced with smaller inter-drop spacing and higher drop widths have regions
of unreduced material that can be seen in black (D). Ends of lines will naturally have slightly more
material due to the ends being pulled in due to capillary action. As seen in B, the small amount of
additional material at the end of the lines pushes the feature height up enough that unreduced
segments remain as seen in black on the right edge of (B). Scale bars 500 μm.

5.3.4 N2 Plasma Reduction

Lines for which pressure cycling was performed to ensure higher N 2 atmosphere

and remove O2 (Section 4.3.3, Figure 4-11) produced dark/grey black material,

indicating issues with reduction of the Pt ink. These samples immediately

delaminated upon coming into contact with water drops (Figure 5-5A). Pressure

cycling takes roughly 90 s, during which time enough EG may have evaporated to

impede reduction. Lines for which the plasma chamber was turned on immediately

upon insertion of the sample, produced highly reflective metallic structures (Figure

5-5B), the same as those observed in the conductivity images in Chapter 4 section

4.3.5. Adhesion of Pt to the substrate was also demonstrated as there was no

noticeable degradation/delamination after 10 min of sonication in water.

5.3.5 Conductivity

Using a multimeter, we demonstrated that current was able to be passed over 2cm

of line, however the readings were transient. Moving the leads closer together

along the line would reinitiate current, before stopping again. We attribute this to

the fact that the PDMS is more deformable than the Pt. Each time the leads were

manually pressed against the Pt, it caused the lines to crack as the PDMS

Ryan P. Sullivan 189

underneath the lines deformed and stretched. These cracks were evident using

optical profilometry immediately after the attempts at measuring conductivity were

performed (not shown).

Conductive-AFM was attempted but was also unsuccessful. Neither direct contact

with the lines nor indirect contact using copper tape as an intermediate connector

produced a current. This may be due to the cracks caused by the PDA films which

form islands that are too small to bias and measure directly (Figure 5-7).

5.3.6 Polydopamine film

While a concentration of 1 mg/mL with a 60 min synthesis resulted in PDA films

which were adequate for depositing lines with minimal blebbing, the PDA film

started to show cracks (Figure 5-7). We attribute this to dehydration and

subsequent contraction of the film as evidenced by recent literature (Zhang et al.,

2013), which would be accelerated by the vacuum conditions of plasma treatment.

Additionally the mechanical mismatch between the glassy PDA and soft PDMS can

cause cracks in the substrate (Yang and Zhao, 2011).

Ryan P. Sullivan 190

Figure 5-7: AFM Images of PDMS-PDA surface (A, B), PDMS-PDA having undergone 10 min N2 plasma
treatment (C); Reduced Pt on PDMS-PDA after 10 min N2 plasma treatment (D). Panel A shows the
homogenous nature of the PDA film, while the zoomed in image B shows the nanoporous nature of
the PDA film, agreeing with images in literature (Zhang et al., 2013). Panel C shows changes in the
morphology of the PDA film now covered in cracks. This is attributed to dehydration of the PDA film
due to the vacuum of plasma treatment (Zhang et al., 2013). Panel D shows the platinum line on the
PDMS-PDA substrate has cracked and formed islands breaking conductivity. Scale bars: (A) 10 μm,
(B) 2 μm, (C) 10 μm, (D) 10 μm.

5.3.7 Medical Readiness

The issues of delamination and potential effects byproduct were discussed in the

previous chapter, section 4.4. While the data in this chapter has not affected the

outlook of delamination, the surface modification, particularly plasma treatment of

the PDMS surface along with polydopamine modification, is an important aspect to

consider. Plasma treatment has been known to change the PDMS surface both

Ryan P. Sullivan 191

chemically and physically. It can become roughened and stiffer, increasing both

crystallinity and hydrophilicity (Chen 2017, Phan et al, 2017). The increased rigidity

and crystallinity may improve neurite outgrowth and cell movement (Lo et al.,

2000, Leach et al., 2007). The hydroxyl motifs may increase neuronal adhesion

while decreasing fibroblast adhesion as previously shown (Green et al., 2014), but

may also be short lived depending on the crosslinking (Hillborg et al., 2004).

Previous studies have shown however that PDMS samples that underwent plasma

treatment maintained their hydroxyl presentation and increase hydrophilicity when

immediately immersed in polar media.

Dopamine is a neurotransmitter used in many parts of the brain. PDA films have

been shown to interact with neuronal cell cultures positively as coatings for

cochlear implants (Schendzielorz et al., 2017) or when coupled with other

molecules such as lysine (Kim et at., 2012, Kang et al., 2011). They have more

generally been shown to improve cell adhesion, serve as bioactive scaffolds while

having long term anti-erosion abilities. (Won et al., 2012, Li et al., 2014) While more

work needs to be undertaken to understand any side effects PDA may present, it is

a promising solution for further modification of implant surfaces.

5.4 CONCLUSION

We have fabricated platinum lines down to widths of 175 μm at speeds of 75

mm/sec (lines 150 mm long, 2 dots wide, in a single layer) using a single jet at 50

μm spacing. To put this in perspective, if all 16 jets of the inkjet cartridge were used

at 3kHz, they would deposit 48,000 features per second; the basic electrode layout

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of the Nucleus CI512 (Cochlear Ltd., 2016) would then take 7.4 s per layer (see 5.S2

for details) to deposit.

While PDA modification of PDMS provided sufficient surface morphologies and

energies to facilitate line printing, further optimization of the PDA synthesis may be

required to prevent the dehydration and cracking from plasma treatment.

Alternatively, localized reduction methods such as laser or photonic sintering may

bypass these morphological changes. Future work should further examine surface

modification strategies for PDMS in more detail in combination with line printing.

Ryan P. Sullivan 193

5.5 APPENDIX

5.5.1 Calculating the effect of pixel resolution on circularity and roundness

To determine the effect of image resolution on circularity and roundness

measurements taken at different magnifications, calibration grids (Figure 5-S1)

were created. Circles with diameters from 1-175 pixels were created using GIMP

(GNU Image Manipulation Program 2.8.10) with a hardness of 1. ImageJ particle

analysis was then used to obtain circularity and roundness values as a function of

the diameter (Figure 5-S2). As the diameter approaches 1, the average circularity

increases approaching unity, though the variability of the measurements increases

dramatically.

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Figure 5-S1: A calibration image of spheres creating using GIMP (GNU Image Manipulation Program)
with diameters from 1 – 139 pixels with a hardness of 1. The calibration image was fed into ImageJ
to determine the effect of pixel size of a perfect circle on the circularity.

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Figure 5-S2: Circularity of circles from calibration image (Figure 5-S1) as a function of diameter in
pixels.

5.5.2 Cochlear CI512 Electrode Calculations

A cochlear implant has 22 platinum electrodes, with leads 800 μm at the base end,

tapering to 500 μm at the apical electrode band (Cochlear Ltd., 2009). There are

three main sections of lengths 60, 13, and 19 mm (Figure 5-S3).

Figure 5-S3: Image of Cochlear implant.

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 60𝑚𝑚 ∗ 9𝑤𝑖𝑑𝑒 13𝑚𝑚 ∗ 9𝑤𝑖𝑑𝑒 19𝑚𝑚 ∗ 8𝑤𝑖𝑑𝑒
𝑇𝑖𝑚𝑒⁡𝑝𝑒𝑟⁡𝑒𝑙𝑒𝑐𝑡𝑟𝑜𝑑𝑒⁡𝑙𝑎𝑦𝑒𝑟 = ( + + )
150𝑚𝑚/𝑠 150𝑚𝑚/𝑠 150𝑚𝑚/𝑠

= 5.4⁡𝑠

5.4𝑠 ∗ 22𝑒𝑙𝑒𝑐𝑡𝑟𝑜𝑑𝑒𝑠
𝑇𝑜𝑡𝑎𝑙⁡𝑡𝑖𝑚𝑒⁡𝑝𝑒𝑟⁡𝑙𝑎𝑦𝑒𝑟⁡(1.1𝜇𝑚) = = 7.4⁡𝑠
16⁡𝑗𝑒𝑡𝑠

800μm⁡thick
𝑇𝑜𝑡𝑎𝑙⁡𝑇𝑖𝑚𝑒⁡(𝑒𝑥𝑙𝑢𝑑𝑖𝑛𝑔⁡𝑃𝐷𝑀𝑆⁡𝑓𝑎𝑏⁡&𝑟𝑒𝑑𝑢𝑐𝑡𝑖𝑜𝑛) = ⁡7.4𝑠 ∗ = 12𝑚𝑖𝑛
1.1μm⁡per⁡layer

5.5.3 Source code for Analyze Stripes Plugin

Below is source code for the ImageJ plugin Analyze Stripes based on the macro by

Justin Bickford (2013). It has been made to be fully automatic with optional pop-ups

for additional user input and is now properly callable from other plugins.

Notepad++ was used to color the code to a custom code set designed to mimic the

Fiji macro builder. It functions as an adjunct to the software covered in Chapter 3.

While the grid analysis software in Chapter 3 provides a way to determine how

reliable ink jetting is in both placement on morphology of droplets, this software is

used to quantitatively determine the quality of line printing patterns. That is,

knowing you can accurately place droplets, what pattern of droplets produces the

best lines with minimal blebbing. The active repository can be found at

https://github.com/RyanPSullivan7/Analyze_Stripes.

Ryan P. Sullivan 197

 1 /*
2 Copyright 2016 Ryan P. Sullivan.
3 Copyright 2013 Justin R. Bickford.
4
5 Permission is hereby granted, free of charge, to any person
6 obtaining a copy of this software and associated documentation files
7 (the “Software”),
8 copy, modify, merge, publish, or otherwise alter this software for
9 educational or academic purposes subject to the following
10 conditions:
11
12 The above copyright notice and this permission notice shall be
13 included in all copies or substantial portions of the Software.
14 The copyright holders of other software modified and included in the
15 Software retain their rights and the licenses on that software
16 should not be removed.
17 Cite the authors (above) of this plugin in any publication that
18 relies on the Software. Also cite those projects on which the
19 Software relies when applicable.
20
21 THE SOFTWARE IS PROVIDED “AS IS”, WITHOUT WARRANTY OF ANY KIND,
22 EXPRESS OR IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF
23 MERCHANTABILITY,
24 FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT
25 SHALL THE AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM,
26 DAMAGES OR OTHER LIABILITY,
27 WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING FROM,
28 OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER
29 */
30
31 import ij.*;
32 import ij.io.*;
33 import ij.gui.*;
34 import ij.measure.Calibration;
35 import ij.measure.Measurements;
36 import ij.measure.ResultsTable;
37 import ij.macro.Interpreter;
38 import ij.plugin.*;
39 import ij.plugin.frame.RoiManager;
40 import ij.plugin.ContrastEnhancer;
41 import ij.plugin.filter.Analyzer;
42 import ij.plugin.filter.ParticleAnalyzer;
43 import ij.process.*;
44 import java.util.ArrayList;
45 import java.util.Arrays;
46 import java.util.List;
47 import java.io.File;
48 import java.io.FilenameFilter;
49 import java.io.IOException;
50 import ij.process.ImageConverter;
51 import ij.process.ImageProcessor;
52 import ij.util.Tools;
53 import ij.gui.PolygonRoi;
54 import ij.gui.PointRoi;

Ryan P. Sullivan 198

 55 import ij.gui.WaitForUserDialog;
56 import ij.gui.Roi;
57
58 public class Analyze_Stripes implements PlugIn {
59
60 protected ImagePlus Image, ActiveImage;
61 ImageProcessor ActiveImageProcessor;
62 ImageConverter ActiveImageConverter;
63 ResultsTable rt;
64 double averageangle, linewidth, RMS_edge_roughness,
65 pkpk_edge_roughness;
66 int nResults;
67 int similarity = 5; // controls how wide to cast the net of
68 similarity
69 int options, measurements;
70 List<Double> group1, group2;
71 double[][] Data;
72 String unit = "pixels";
73 RoiManager ActiveROIManager;
74 Roi[] RoiArray, RoiArray1, RoiArray2;
75 Roi Roi1, Roi2;
76 boolean userInput = false;
77
78 public void run(String arg) {
79 Image=IJ.getImage();//Opened Image
80 ActiveImage=Image.duplicate();//Image on which all the
81 processing is done
82 ActiveImageProcessor = ActiveImage.getProcessor();
83 ActiveImageConverter = new ImageConverter(ActiveImage);
84 unit = ActiveImage.getCalibration().getUnits();
85 ActiveROIManager = new RoiManager(true);
86 rt = new ResultsTable();
87 options = ParticleAnalyzer.CLEAR_WORKSHEET +
88 ParticleAnalyzer.ADD_TO_MANAGER;
89 measurements = Measurements.AREA + Measurements.MEAN +
90 Measurements.STD_DEV + Measurements.MIN_MAX +
91 Measurements.CENTROID + Measurements.ELLIPSE;
92 IJ.run("Set Measurements...", "area mean standard min centroid
93 fit redirect=None decimal=3");
94 //Convert image to binary edges
95 findEdgesAndThreshold();
96 //measure edges
97 ParticleAnalyzer ActiveParticleAnalyzer = new
98 ParticleAnalyzer(options,measurements,rt,0,Double.POSITIVE
99 _INFINITY,0.00,0.10);
100 ActiveImageConverter.convertToGray8();
101 ActiveParticleAnalyzer.setRoiManager(ActiveROIManager);
102 ActiveParticleAnalyzer.analyze(ActiveImage);
103 //Apply rotation to edges
104 calculateAngleAndRotate();
105 //collect Data

Ryan P. Sullivan 199

106 ActiveROIManager.runCommand("Measure");
107 setMeasurementArray();
108 sortDataByMean();
109 mergeROIs();
110 analyzeData();
111 ActiveROIManager.moveRoisToOverlay(Image);//Overlays Edges
112 on original image. Stop prompt from next line and keep the Edges
113 if (linewidth==0){
114 IJ.log("Failed to find distinct edges.");
115 return;
116 }
117 outputToResults();
118 rt.show("Results");
119 return;
120 }
121 //Finds the gradient^4 of the image
122 //converts to pixels > mean*8 to binary and skeletonizes
123 public void findEdgesAndThreshold() {
124 ActiveImageConverter.convertToGray16();
125 ContrastEnhancer enhancer = new ContrastEnhancer();
126 enhancer.equalize(ActiveImage);
127 ActiveImageProcessor = ActiveImage.getProcessor();
128 ActiveImageProcessor.findEdges();
129 ActiveImageProcessor.sqr(); // turns the gradient into
130 gradient squared
131 ActiveImageProcessor.sqr(); // further enhances the good
132 edges
133 enhancer.equalize(ActiveImage);
134 double mean = ActiveImage.getStatistics().mean;
135 double max = ActiveImage.getStatistics().max;
136 ActiveImageProcessor.setThreshold(mean*8,max,ActiveImageP
137 rocessor.OVER_UNDER_LUT);
138 if(userInput)
139 {
140 IJ.run(ActiveImage,"Threshold...","");
141 new WaitForUserDialog("OK").show();
142 if (WindowManager.getWindow("Threshold")!= null){
143 IJ.selectWindow("Threshold");
144 IJ.run("Close");
145 }
146 }
147 IJ.run(ActiveImage,"Convert to Mask","");
148 IJ.run(ActiveImage,"Skeletonize","");
149 return;
150 }
151 //Calculates the area weighted angle and rotates the data using v
152 = x*sin(angle) + y*cos(angle)
153 public void calculateAngleAndRotate(){
154 double cumarea = 0;
155 double sina = 0;
156 double cosa = 0;

Ryan P. Sullivan 200

157 double area, angle;
158 for (int n = 0; n < rt.size();n++) {
159 area = rt.getValueAsDouble(rt.getColumnIndex("Area"),n);
160 angle =
161 2*rt.getValueAsDouble(rt.getColumnIndex("Angle"),n);
162 sina = sina + area*Math.sin(angle*Math.PI/180);
163 cosa = cosa + area*Math.cos(angle*Math.PI/180);
164 cumarea = cumarea+area;
165 }
166 averageangle =
167 Math.abs(0.5*(180/Math.PI)*Math.atan2(sina/cumarea,cosa/cu
168 marea)); // this is the area weighted average angle
169 // rotate the data
170 IJ.run(ActiveImage,"Select All","");
171 ActiveImageConverter.convertToGray32();
172 IJ.run(ActiveImage,"Macro...", "code=[v=
173 x*sin(PI/180*"+averageangle+")+y*cos(PI/180*"+averageangle+")
174 ]");
175 return;
176 }
177 //Grabs the mean, standard deviation, max, and min values of the
178 edges and puts them in Data[][]
179 public void setMeasurementArray(){
180 nResults = rt.size();
181 Data = new double[4][nResults];
182 for(int i =1; i<nResults; i++) {
183 Data[0][i]=i;
184 Data[1][i]=rt.getValueAsDouble(rt.getColumnIndex("Mean")
185 ,i);
186 Data[2][i]=rt.getValueAsDouble(rt.getColumnIndex("StdDev
187 "),i);
188 Data[3][i]=rt.getValueAsDouble(rt.getColumnIndex("Max"),
189 i)-rt.getValueAsDouble(rt.getColumnIndex("Min"),i);
190 }
191 return;
192 }
193 //Sort Data[][] by mean
194 public void sortDataByMean() {
195 for(int i = 1; i<nResults-1; i++){
196 double tempIndex = Data[0][i];
197 double tempMean = Data[1][i];
198 double tempStdDev = Data[2][i];
199 double tempextremea = Data[3][i];
200 int j = i - 1;
201 while (j>=0 && Data[1][j] > tempMean){
202 Data[0][j+1]=Data[0][j];
203 Data[1][j+1]=Data[1][j];
204 Data[2][j+1]=Data[2][j];
205 Data[3][j+1]=Data[3][j];
206 }
207 Data[0][j+1]=tempIndex;

Ryan P. Sullivan 201

208 Data[1][j+1]=tempMean;
209 Data[2][j+1]=tempStdDev;
210 Data[3][j+1]=tempextremea;
211 }
212 return;
213 }
214 //Filter and Merge ROIs (edges) to only 2 outer most edges
215 public void mergeROIs() {
216 group1 = new ArrayList<Double>();
217 group1.add(Data[0][0]);
218 for (int i = 1; i < (nResults-1);i++) {
219 if ((Data[1][i]-similarity*Data[2][i]) <
220 (Data[1][0]+similarity*Data[2][0]) ) { // if the lines
221 are
222 similar to the minimum line, add them to group1
223 group1.add(Data[0][i]);//addes new element to group 1
224 with value of roiindex[i]
225 }
226 }
227 int[] group1a = new int[group1.size()];
228 for(int i =0; i < group1.size(); i++){
229 double temp = group1.get(i);
230 group1a[i] = (int)temp;
231 }
232 group2 = new ArrayList<Double>();
233 group2.add(Data[0][nResults-1]);
234 for (int i=(nResults-2);i>1;i--) {
235 if ((Data[1][i]+similarity*Data[2][i]) >
236 (Data[1][nResults-1]-similarity*Data[2][nResults-1]) ) {
237 // if the lines are similar to the maximum line, add them to
238 group2
239 group2.add(0,Data[0][i]);
240 }
241 }
242 int[] group2a = new int[group2.size()];
243 for(int i =0; i < group2.size(); i++){
244 double temp = group2.get(i);
245 group2a[i] = (int)temp;
246 }
247 int count;
248 //IJ.run("Select None");
249 ActiveROIManager.deselect();
250 ActiveROIManager.setSelectedIndexes(group1a);
251 if(group1a.length > 1) {
252 ActiveROIManager.runCommand(ActiveImage,"Combine");
253 //Creates new ROI that is combination of group 2
254 count = ActiveROIManager.getCount();
255 Roi1 = ActiveROIManager.getRoi(count-1);//Selects the
256 combined group1 ROI
257 }
258 else{
259 Roi1 = ActiveROIManager.getRoi(group1a[0]);

Ryan P. Sullivan 202

260 }
261 ActiveROIManager.setSelectedIndexes(group2a);
262 if(group2a.length > 1) {
263 ActiveROIManager.runCommand(ActiveImage,"Combine");
264 //Creates new ROI that is combination of group 2
265 count = ActiveROIManager.getCount();
266 Roi2 = ActiveROIManager.getRoi(count-1); //Selects the
267 combined group2 ROI
268 }
269 else{
270 Roi2 = ActiveROIManager.getRoi(group2a[0]);
271 }
272 ActiveROIManager.reset();
273 rt.reset();
274 ActiveROIManager.add(ActiveImage,Roi1,0);
275 ActiveROIManager.select(ActiveImage,0);
276 Analyzer ActiveAnalyzer = new
277 Analyzer(ActiveImage,measurements,rt);
278 ActiveAnalyzer.measure();
279 ActiveROIManager.add(ActiveImage,Roi2,1);
280 ActiveROIManager.select(ActiveImage,1);
281 ActiveAnalyzer.measure();
282 ActiveROIManager.runCommand(Image,"Show All without
283 labels");// removes the labels, which a user may find confusing
284 in the context of this macro
285 return;
286 }
287 //Calculate Linewidth, RMS edge roughness, and PkPk edge roughness
288 public void analyzeData(){
289 linewidth = rt.getValue("Mean", 1) - rt.getValue("Mean",
290 0);
291 RMS_edge_roughness =
292 Math.sqrt((Math.pow(rt.getValue("StdDev", 1), 2) +
293 Math.pow(rt.getValue("StdDev", 0), 2))/2); // this is the
294 correct form
295 if(rt.getValue("Max",1) - rt.getValue("Min", 1) >
296 rt.getValue("Max", 0) - rt.getValue("Min", 0)){
297 pkpk_edge_roughness = rt.getValue("Max", 1) –
298 rt.getValue("Min", 1);
299 }
300 else{
301 pkpk_edge_roughness = (rt.getValue("Max", 0) –
302 rt.getValue("Min", 0));
303 }
304 return;
305 }
306 public void setUserInput(boolean userInput){
307 this.userInput = userInput;
308 }
309 public void outputToResults(){
310 rt = new ResultsTable();
311 rt.incrementCounter();

Ryan P. Sullivan 203

312 rt.addLabel("Average Angle");
313 rt.setValue(1,0,averageangle);
314 rt.setValue(2,0,"degrees");
315 rt.incrementCounter();
316 rt.addLabel("Line width");
317 rt.setValue(1,1,linewidth);
318 rt.setValue(2,1,unit);
319 rt.incrementCounter();
320 rt.addLabel("RMS Edge Roughness (Rq)");
321 rt.setValue(1,2,RMS_edge_roughness);
322 rt.setValue(2,2,unit);
323 rt.incrementCounter();
324 rt.addLabel("Peak-to-Peak Edge Roughness (Rt)");
325 rt.setValue(1,3,pkpk_edge_roughness);
326 rt.setValue(2,3,unit);
327 return;
328 }

Ryan P. Sullivan 204

 CHAPTER SIX: CONCLUSIONS AND
OUTLOOK

6.1 OVERALL FINDINGS

The series of studies presented in this thesis have for the first time demonstrated

the potential of platinum polyol inks for use in additive fabrication on flexible,

insulating, and biocompatible substrates for medical implants, and have laid the

foundation for deposition control parameters for future studies. The main findings

from these studies are outlined below.

Chapter 2 showed that the approach and withdrawal speeds of DPN cantilevers

significantly affect the deposition of polar inks, specifically H2PtCl6 in EG, on to the

hydrophobic substrate PDMS. Line printing of this ink onto PDMS required a

reactive printing approach using a polydopamine surface modification to react with

the H2PtCl6 in the ink as the tip passed along the path of the line. Plasma treatment

was found to coalesce the particle morphology of lines to continuous flat surfaces.

The line deposition rate and control were found not to be sufficient for large scale

fabrication of electrode leads.

Chapter 3 provides a new analytical method and accompanying software plugin to

overcome the limited existing optics systems for monitoring inkjet drop formation

Ryan P. Sullivan 205

and flight. This ImageJ plugin was demonstrated to be able to automatically filter,

detect, and analyze optical microscopy images of deposited features at 13μs

timescale, allowing for the analysis of the very large datasets used in the

development of inkjet waveforms in Chapters 4 and 5.

Chapter 4 explored the development of inkjet waveforms which allowed for the

rapid deposition of 48,000 platinum features per second with diameters of 22.89 ±

4.79 μm using 16 jets. It was found that the time between deposition and reduction

of the ink plays a critical role in the success of the final reduction. Samples reduced

immediately after deposition (within a 30 s timeframe from the deposition of the

first feature, to the beginning of the reduction phase), had high conductivities

which were comparable to those of bulk platinum, 78 nΩ*m vs 105 nΩ*m. Samples

which were delayed however (i.e. 20 to 30 minutes between the deposition of the

feature and its subsequent reduction), did not fully reduce, and had significantly

lower conductivities as well as regions of unreduced cubic salts.

Chapter 5 translated the inkjetting technique used in Chapter 4 from 10 pL to 1 pL

volumes, allowing for the deposition of 48,000 platinum features per second with

diameters of 10.08 ± 2.21 μm using 16 jets. The high hydrophobicity of PDMS was

found to cause the overlapping features to bead up into discrete spherical caps

rendering line printing untenable. While surface modification of PDMS by plasma

was not successful in allow for the controlled deposition of lines, 1 hour of

polydopamine modification provided a suitable surface energy and chemistry for

successful deposition of platinum lines. While 1 pL jets were found to offer double

Ryan P. Sullivan 206

the resolution of 10 pL jets, the 10 pL jets were ultimately used to deposit platinum

lines due to their higher possible jetting frequency. Higher jetting frequencies

reduce the total printing time, which as shown in Chapter 4 was found to be the

limiting factor on the successful reduction of the ink. Ultimately, 175 μm wide

platinum lines were able to be reproducibly deposited at rates of 75 mm/s using 16

jets at 10 kHz. The polydopamine film was found to contract as it dehydrated from

plasma treatment, ultimately leading to the formation of islands in the deposited

lines.

The following section will be a general discussion of the main findings of this thesis,

demonstrating how it fits into existing literature. As detailed discussions of the

results were included in Chapters 2-5, only the data most pertinent to the future

directions of this additive fabrication technique will be addressed.

6.2 UPSCALING AND DOWNSCALING POLYOL INKS

Polyol inks have been shown to produce reduced individual features ranging from

as low as 50 nm as demonstrate in our previous work (O’Connell et al., 2014), up to

20 μm in the current work presented in this thesis using both DPN and Inkjet

printing. Cells are functionally nanomachines, and the nanoscale control of the

topography and surface chemistry of an implant is important in order to be able to

control the response of the tissue surrounding the implant. However, implants are

fundamentally macroscale objects, and can be up to centimeters in size. This means

that additive fabrication techniques need to be able to be both scaled down so that

the individual feature size approaches the nanometer and submicron ranges so that

Ryan P. Sullivan 207

it can interface directly with individual cells, while also being able to be scaled up so

that throughput can be increased in order to cover the centimeter range sizes that

would be required for an entire implant. Both previous work (O’Connell et al., 2013;

Wagner et al., 2013) and the work contained in Chapter 2 of this thesis, have shown

that DPN can be used to print conductive materials used in implants; however DPN

as a printing technique is fundamentally limited in its ability to be upscaled. While

related techniques like polymer pen lithography are able to scale up the number of

individual tips used, they are limited to the use of predefined stamps or identical

rows of DPN cantilevers depositing identical parallel patterns. The lack of true

upscaling of multiple individually addressable DPN tips is part of the reason that

NanoInk, the largest manufacturer of DPN machines, tips, and consumables, shut

down in February 2013 in the early phases of this PhD (Johnson, 2013).

Conversely, inkjet printing has been shown to be highly scalable, with each jet being

individually addressable. With resolution ranges of 100 nm – 50 μm, and firing rates

of 1-20 kHz, inkjet and Super Inkjet Technology (SIT) are primed for fabrication at

full implant scales. By far, the limiting factor time-wise for the deposition of polyol

inks is the plasma reduction. A series of lines takes only seconds to deposit, but the

subsequent plasma treatment used may take 10 min or more, and is limited in its

penetration depth. Additionally plasma treatment affects the substrate as a whole.

The introduction of additional components to the printer, such as local laser

systems, will allow increased throughput with polyol inks by allowing reduction to

be conducting in the same machine. Additionally, work is already being done using

inkjetting as a technique for printing the other components of neural implants such

Ryan P. Sullivan 208

as PDMS and cell lines (Ferris et al., 2013a, 2013b; Stieghorst et al., 2016; Yamazaki

et al., 2007). Using targeted laser systems, inkjetted PDMS precursors have been

shown to be able to be annealed in 2 s without significantly increasing the

temperature of the surrounding substrate (Stieghorst et al., 2016). Combining this

existing technique with the polyol inks developed in our previous work and the

work contained within this thesis, may allow for the fabrication of complete devices

such as a retinal or cochlear implant, both of which may consist of PDMS and Pt.

Future formulations of polyol inks may be developed to have control of the acidity,

which would result in reduced degradation of the inkjet heads, as well as influence

the nucleation and growth rates, which are known to be affected by –OH species

(Sun et al., 2010). Additionally, combined metal inks, in particular H2PtCl6 and

H2IrCl6, would allow deposition of alloys which are highly desirable for use in

medical implants. Furthermore, combinations of alcohols such as ethanol or

glycerol may be added to ink formulations in order to alter the rheology of the ink

for different print head geometries, or to allow for longer stabilities of the ink on

the substrate.

In conclusion, the work presented in this thesis contributes to the ever growing

body of knowledge of additive fabrication techniques that can be used to deposit

metal for custom neural implants. Further work on the reduction will be required in

order to overcome the observed limitations in plasma penetration depth and to

increase the rate of reduction as it is the limiting factor time-wise, including altering

the acidity and –OH species as well as tight thermal control from targeted

Ryan P. Sullivan 209

stimulation. Additionally, combining polyol inks with existing silicone and hydrogel

inks may allow fabrication of neural implant prototypes with a single inkjet printer.

Ryan P. Sullivan 210

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