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Multi-Panel On-Single-Chip Memristive Biosensing
Multi-Panel On-Single-Chip Memristive Biosensing
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5770 IEEE SENSORS JOURNAL, VOL. 19, NO. 14, JULY 15, 2019
techniques. This biosensing scheme paves the way for indi- B. Multiplexing Approach for Sensing
vidualized biomarker sensing in a complex matrix, as well
A single-chip, detection in a multi-panel mode is for
as for multiple biomarkers/drugs detection on the same chip
the first time attempted implementing memristive biosen-
and application in theranostics. Sensing chips of memristive
sors. The memristive devices included in the same 1 cm ×
biosensors conjugated with metallic extension electrodes are
1 cm chip are first subjected to a silanization step with
validated for successful sensing performance and then imple-
GPTES (440167, Sigma-Aldrich). Then the devices are sep-
mented for multi-panel sensing.
arately bio-functionalized with different antibody solutions.
Antibodies either from mouse, goat or rabbit were diluted
II. M ATERIALS AND M ETHODS
within PBS to get a final concentration of 0.5 mg/ml with
A. Memristive Biosensing Chips 0.5% trehalose (T9449, D-(+)-Trehalose dihydrate from corn
Two-terminal silicon (Si) nanofabricated wire-arrays starch = 99%, Sigma). Multiplexed biosensing requires post-
exhibiting memristive electrical properties are acquired fabrication nanowire functionalization with probe molecules,
through a top-down fabrication process using Silicon-on- (though robotic spotting [24], ink-jet printing [25], selec-
Insulator (SOI) wafers as described in [7], by implementing tive heating [26], electrochemically [27], or via dip-pen
electron beam lithography and Deep Reactive Ion Etching nanolithography [28]). In this work, a micro-spotter (GeSim
(DRIE). The wires are suspended and anchored between NanoPlotter 2.1) is used for specific and separate bio-
Nickel Silicide (NiSi) pads forming Schottky-barriers and functionalization of each nanodevice with the different anti-
serving for the electrical characterization of the nanodevices. bodies under consideration, with a drop of 400 pL antibody-
Metal lines are integrated on top of the already fabricated solution of an initial diameter set at 150 µm. A rabbit antibody
nanostructures and serve as extension line electrodes to the is used as a positive-control bio-functionalization reagent and
NiSi pads of the devices, as described in [9]. The metal is spotted to various devices of the sensing chip. Meanwhile,
lines are designed taking into consideration the fact that three different negative controls are taken into consideration:
the hysteretic properties of the memristive biosensors are a mouse and a goat antibody both applied at the same
sensitive to additional resistive elements introduced in series concentrations as the positive control as well as plain buffer
to the nanofabricated devices, eliminating or even completing solution (PBS).
masking the memristive properties of the nanodevices. For The micro-spotting procedure finally results in discrete
this reason, the extension lines design follows the outcomes of bio-functionalization of different devices (Fig. 1). Following
a relevant computational study and calculations [9], [13] for the bio-functionalization the chip remains in a humid
respecting the resistance tolerance of the system. A common chamber for 1 h in order to ensure the attachment of the
source and separated drains are considered for individual bio-functionalization regents on the nanodevice surface. The
measurement of twelve integrated sensors (as seen in Fig. 1) additional humidity introduced, aims at the conservation of
that allow possibility for adequate statistical analysis to the liquid antibody-solution drops which are very sensitive to
determine average value and standard deviation. This evaporation issues. In order to prevent non-specific binding
statistical analysis is required for obtaining reliable results of proteins during the detection measurement, the remaining
and overcome the variability of the sensors originating from active GPTES-derived groups are passivated by applying
the DRIE process. In addition, such realization of multiple 10 mM ethanolamine in PBS solution at room temperature.
sensors allows the possibility for simultaneous sensing (multi- Washing process is further performed to remove unreacted
panel sensing), that consists the main aim of the present study. molecules.
To confirm the sound functionality of the metal lines the An additional blocking step is carried out with PBS
memristive devices were functionalized by exposure of the containing 3% gelatin from cold water fish skin. This further
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TZOUVADAKI et al.: MULTI-PANEL, ON-SINGLE-CHIP MEMRISTIVE BIOSENSING 5771
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5772 IEEE SENSORS JOURNAL, VOL. 19, NO. 14, JULY 15, 2019
Fig. 5. Illustration of the memristive bio-sensing platform, implemented for theranostics in combination with specially designed microfluidic circuit.
binding (Fig. 4). However, no significant difference is recorder designed memristive sensing chips offering disposable sens-
for the case of the negative-control bio-functionalized devices ing modules by integrating memristive wires and extension
(Fig. 4). Further study of the analytical performance of the metal electrodes are validated for successfully providing the
nanodevices at the different antigen steps, demonstrates a electrical characteristics of the devices before and after the
decreasing behavior only for the nanodevices functionalized bio-functionalization and the target uptake steps. Most impor-
with the positive control antibody while the voltage gap for the tantly, this novel sensing paradigm demonstrates successful
other nanodevices remains constant at the levels of the values implementation of the memristive biosensors in a multi-panel
after the bio-functionalization. This finding brings proof of sensing scheme in both labeled and label-free sensing. The
the possibility for efficient individualized bio-functionalization system presented in this work paves the way for advanced
of different nanodevices on the same chip. Moreover, these bio-detection with memristive biosensors, allowing individu-
results demonstrate the capability of the memristive biosensors alized detection of specific targets in complex matrixes in the
to provide discrete sensing of a target molecule against other presence of non-specific molecules or in even more complex
non-specific molecules through a tailored functionalization of schemes i.e. cells, tumor extracts, holding also great promise
individual devices. Therefore, this study consists a very impor- for fast and ultrasensitive sensing in theranostics.
tant step towards future implementation of the memristive
biosensors for the detection of a specific biomarker or drug
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Authorized licensed use limited to: University of Gent. Downloaded on November 17,2022 at 08:24:12 UTC from IEEE Xplore. Restrictions apply.
TZOUVADAKI et al.: MULTI-PANEL, ON-SINGLE-CHIP MEMRISTIVE BIOSENSING 5773
[8] F. Puppo, F. L. Traversa, M. Di Ventra, G. De Micheli, and S. Carrara, Ioulia Tzouvadaki received the B.Sc. degree in
“Surface trap mediated electronic transport in biofunctionalized silicon physics from the National and Kapodistrian Uni-
nanowires,” Nanotechnology, vol. 27, no. 34, 2016, Art. no. 345503. versity of Athens (UOA), Greece, the M.Sc. degree
[9] S. Naus, I. Tzouvadaki, P.-E. Gaillardon, A. Biscontini, G. De Micheli, in microsystems and nanodevices from the National
and S. Carrara, “An efficient electronic measurement interface for Technical University of Athens (NTUA), and the
memristive biosensors,” in Proc. IEEE Int. Symp. Circuits Syst. (ISCAS), Ph.D. degree from EPFL, Switzerland. During her
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[10] I. Tzouvadaki, A. Tuoheti, G. De Micheli, D. Demarchi, and S. Carrara, Room Laboratory of the National Center for Sci-
“Portable memristive biosensing system as effective point-of-care device entific Research (NCSR) Demokritos in the field
for cancer diagnostics,” in Proc. IEEE Int. Symp. Circuits Syst. (ISCAS), of experimental construction processes concerning
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[11] F. Puppo, M. Di Ventra, G. De Micheli, and S. Carrara, “Memris- zation process of nanomaterials and nanodevices. Her M.Sc. thesis concerned
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pp. 76–79, Jun. 2014. als, within the Computational Materials Science and Engineering (CoMSE)
[12] F. Puppo, M.-A. Doucey, M. Di Ventra, G. De Micheli, and S. Carrara, Research Group, School of Chemical Engineering, NTUA. Her Ph.D. work
“Memristor-based devices for sensing,” in Proc. IEEE Int. Symp. Circuits with the Integrated System Laboratory (LSI) of EPFL, focused on the
Syst. (ISCAS), Jun. 2014, pp. 2257–2260. fabrication and characterization of nanostructures and their implementation
as ultrasensitive nano-bio-sensors in both diagnostics and therapeutics.
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Abuduwaili Tuoheti was born in Xinjiang, China,
[15] A. Vallero et al., “Memristive biosensors integration with microfluidic in 1984. He received the B.Sc. degree in micro-
platform,” IEEE Trans. Circuits Syst. I, Reg. Papers, vol. 63, no. 12, electronic physics from Nanjing University, China,
pp. 2120–2127, Dec. 2016. in 2008 and the M.Sc. degree from the Politecnico
[16] C. Yang, C. Xu, X. Wang, and X. Hu, “Quantum-dot-based biosensor di Torino in 2014. He was a Teacher with the
for simultaneous detection of biomarker and therapeutic drug: First Shufu High School, China, from 2008 to 2011.
steps toward an assay for quantitative pharmacology,” Analyst, vol. 137, He was an Embedded Systems Engineer with ST
no. 5, pp. 1205–1209, 2012. Microelectronics in 2014. Then, he was a Software
[17] S. Carrara, A. Cavallini, V. Erokhin, and G. De Micheli, “Multi-panel Developer with the National Research Council of
drugs detection in human serum for personalized therapy,” Biosensors Italy from 2015 to 2016. He is currently a Doc-
Bioelectron., vol. 26, pp. 3914–3919, May 2011. toral Assistant with the Department of Electronics
[18] M. Lee, K. Y. Baik, M. Noah, Y.-K. Kwon, J.-O. Lee, S. Hong, and Telecommunications, Politecnico di Torino, under the supervision of
“Nanowire and nanotube transistors for lab-on-a-chip applications,” Lab D. Demarchi and S. Carrara. His main research interests include embedded
Chip, vol. 9, no. 16, pp. 2267–2280, 2009. intelligent system design for bio-medical applications, analog front-end design
[19] Y. Zhang, R. Chen, L. Xu, Y. Ning, S. Xie, and G.-J. Zhang, “Silicon for bio-sensors, machine-learning based algorithm development for sensor data
nanowire biosensor for highly sensitive and multiplexed detection of processing, and cloud-based software development for healthcare.
oral squamous cell carcinoma biomarkers in saliva,” Anal. Sci., vol. 31,
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[20] M. Khairy, B. G. Mahmoud, and C. E. Banks, “Simultaneous deter-
mination of codeine and its co-formulated drugs acetaminophen and
caffeine by utilising cerium oxide nanoparticles modified screen-printed
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[21] M. Khairy, A. A. Khorshed, F. A. Rashwan, G. A. Salah, Séverine Lorrain received the Ph.D. degree in plant
H. M. Abdel-Wadood, and C. E. Banks, “Simultaneous voltammetric science from the University of Toulouse, France,
determination of antihypertensive drugs nifedipine and atenolol utilizing for the work centered on the molecular pathways
MgO nanoplatelet modified screen-printed electrodes in pharmaceuticals deregulated in an Arabidopsis lesion mimic mutants,
and human fluids,” Sens. Actuators B, Chem., vol. 252, pp. 1045–1054, in 2004. She then moved to the University of
Nov. 2017. Lausanne in the Group of Prof. C. Fankhauser to
[22] P. N. Manikandan and V. Dharuman, “Electrochemical simultaneous decipher the molecular pathways involved on the
sensing of melatonin, dopamine and acetaminophen at platinum doped plant responses to their changing light environment.
and decorated alpha iron oxide,” Electroanalysis, vol. 29, no. 6, Since 2016, she has been in charge of the Reverse
pp. 1524–1531, 2017. Phase Protein Array platform at the Protein Analysis
[23] C. Wang, Z. Sun, L. Ma, and M. Su, “Simultaneous detection of Facility of the University of Lausanne.
multiple biomarkers with over three orders of concentration differ-
ence using phase change nanoparticles,” Anal. Chem., vol. 83, no. 6,
pp. 2215–2219, 2011.
[24] R. P. Auburn, D. P. Kreil, L. A. Meadows, B. Fischer, S. S. Matilla, and
S. Russell, “Robotic spotting of cDNA and oligonucleotide microarrays,”
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Manfredo Quadroni graduated with a degree in
[25] A. Bietsch, J. Zhang, M. Hegner, H. Lang, and C. Gerber, “Rapid
biochemistry and then received the Ph.D. degree
functionalization of cantilever array sensors by inkjet printing,” Nan-
from ETH Zürich for work centered on calcium-
otechnology, vol. 15, no. 8, pp. 873–880, 2004.
mediated signaling and protein phosphorylation,
[26] I. Park, Z. Li, A. P. Pisano, and R. S. Williams, “Selective surface where he also received his training in proteomics in
functionalization of silicon nanowires via nanoscale joule heating,” Nano the laboratory of Dr. P. James. During a post-doctoral
Lett., vol. 7, no. 10, pp. 3106–3111, 2007. stay at the University of British Columbia, Canada,
[27] Y. L. Bunimovich, G. Ge, K. C. Beverly, R. S. Ries, L. Hood, he also became familiar with signal transduction in
and J. R. Heath, “Electrochemically programmed, spatially selective immunology. A second post-doctorate brought him
biofunctionalization of silicon wires,” Langmuir, vol. 20, no. 24, back to ETH Zürich, where he continued working on
pp. 10630–10638, 2004. the development of methods for proteome analysis.
[28] K. Salaita, Y. Wang, and C. A. Mirkin, “Applications of dip-pen He joined the University of Lausanne in 2000, where he has been in charge
nanolithography,” Nature Nanotechnol., vol. 2, pp. 145–155, Feb. 2007. of the setup and operation of the Protein Analysis Facility since 2002.
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5774 IEEE SENSORS JOURNAL, VOL. 19, NO. 14, JULY 15, 2019
Marie-Agnès Doucey received the Engineering Danilo Demarchi (M’10–SM’13) received the mas-
degree in biotechnology from the Ecole Supéerieure ter’s and Ph.D. degrees in electronics engineering
de Biotechnologies de Strasbourg (ESBS), France, from the Politecnico di Torino (Polito), Italy, in 1991
and the Ph.D. degree in biochemistry from and 1995, respectively. He is an Associate Professor
the Friedrich-Miescher Institute, Novartis, Basel, with Polito, where he is leading the Micro and Nano
Switzerland. She is currently a Project Leader with Electronic Systems Group, Department of Electron-
the Ludwig Institute for Cancer Research, University ics and Telecommunications. He is also coordinating
of Lausanne. She has more than 20 publications the microelectronics research line at the Center for
in peer-reviewed international journals. Her cur- Space Human Robotics, Istituto Italiano di Tecnolo-
rent work focuses on the tumor microenvironment gia, Polito. He has authored and co-authored two
and the role of monocytes in breast cancer tumor patents and over 150 scientific publications.
development.
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