Laboratory of Biosensors

and Bioelectronics

Biosensors and Bioelectronics

Janos (Vörös), Tomaso (Zambelli)

Laboratory of Biosensors and Bioelectronics

Institute for Biomedical Engineering,

voros@ethz ch zambelli@ethz.ch
voros@ethz.ch, zambelli@ethz ch

http://lbb.ethz.ch/

Phones:
Janos: +41 44 632 59 03
Tomaso: +41 44 632 4575
 Program of Today’s Class

• Goal of the lecture

g
• Program plan
p for the semester
• Basic definitions
• Biosensing:
– Concept, Applications, Sensitivity
• Bioelectronics:
– Applications,
Applications Some history
history, Electrolysis
• Discussions
 Goal of the Lecture

You will
• learn the motivations behind biosensing and
bioelectronics
• learn the basic concepts in biosensing and
bioelectronics
• be able to solve typical problems in biosensing and
bioelectronics
• learn to locate information fast
 Definitions

Biosensor:
1. A device that detects, records, and transmits information
regarding a physiological change or process.
2 A d
2. device
i that
h uses bi
biological
l i l materials
i l to monitor
i the
h presence
of various chemicals in a substance.

Bioelectronics:
1 The application of the principles of electronics to biology and
1.
medicine.
y of
2. The study f the role of
f intermolecular
m electron transfer
f in
physiological processes.

Source: http://www.thefreedictionary.com
 P bl
Problems
Solved with Biosensors
Today
 Glucose Monitoring

From a drop of blood Implantable

 High Throughput Analysis

Diagnostics Drug-
Drug-discovery

>800 blood tests per hour >1000 microtiter plates per day
 Biosensors
os nsors in
n Everyday
E ry ay Life
Lf

Hospitals
– 8000 samples
– p
per hour

Home
– 1 sample
– within 5 s
OWLS Biosensors for Research DWI

http://www.microvacuum.com
p http://www.farfield-sensors.co.uk

ZeptoReader QCM--D
QCM

http://www.zeptosens.com http://www.q-sense.com
 Cell–based Sensing

Neurons--on-
Neurons on-a-chips Multi-
Multi-channel recording

Physiology of brain tissue slices

 DNA Microarrays

Electronic DNA Array (Nanogen) DNA chip (Affymetrix)

 Nanobiosensors

Nanowire Arrays
y Ion--channel based sensing
Ion

Nat. Biotechnol. 23, 1294-1301 (2005) Nature 387, 580-583 (1997)

 Main Players on the Market

Total volume in 2006 ~7 billion US$

 Bio-
Re
ecogn sample
nition
n Elem
ment

Trransd
ducerr

Signa
S al Pro
ocesssing
Biosensor Principle

Signal
 Biosample (Proteins)

Bio-
sample
 What is in Blood?
Total protein = 50 mg/mL
Interesting
g proteins
p <1 ng/mL
g
 Recognition Elements

ment
n Elem
Bio-
sample nition
ecogn
Re
 Suitable Immobilization of Receptors

Biotin NTA-Ni2+-6his

IgG IgG
Oriented receptor
his-tag
his tag
Suitable linker
Low non-specific binding
Ni
PEG-NTA
NTA
Stable binding

Sensor Surface Sensor Surface

 Bio-
Re
ecognsample
nition
n Elem
ment

Trransd
ducerr
Transducers
 Transducers by Signal Type

• Radioactivity
p
• Optical
– waveguides, ellipsometry, SPR, fluorescence, …
• Electrical
– enzymatic, amperometric, potentiometric,…
• Mechanical
– Quartz crystal microbalance,
microbalance SAW device,
device
cantilever devices, …
 Bio-
Re
ecogn sample
nition
n Elem
ment

Trransd
ducerr

Signa
S al Pro
ocesssing
Signal Processing
 Genomics

3 1 billi
3.1 billion
n b
bases
30’000 genes

Proteins, their function

and connections
-> Proteomics

-> High Throughput

Screening
Summary of Biosensing Concept
 Sensitivity of a Biosensor

Source:
Roger P. Ekins
Clinical Chemistry 44(9):2015-2030 (1998)
– See pdf at the course web-site
 “Langmuir”
Langmuir Isotherm

1200

1000

Equilibrium Values
800

Model: Langmuir
Mmax 1081 ±60.7011
600
K 38 ±7.28906

400

E
200

0
0 100 200 300 400 500 600 700
Conc [nM]
Conc.
Bi l t
Bioelectronics
i
 Definition

Bioelectronics:
1. The application of the principles of electronics to biology and
medicine.
2 Th
2. The study
d of
f the
h role
l of
f iintermolecular
l l electron
l transfer
f iin
physiological processes.

Source: http://www.thefreedictionary.com
 Applications of Bioelectronics

Biochips (Biosensors)
Implantable Medical Devices
Prosthetic Devices ((Hearing
g Aids,, Artificial Sight,
g , Limb Replacement)
p )
Artificial Organs
Electronic Pills
Surgical and Medical Devices
Biofuel Cells
Molecular Motors
Molecular Electronics
Biofabrication Templates

h
http://www.bioelectronicscorp.com/
// bi l i /
http://www.sric-bi.com/Explorer/NGT-BE.shtml
Some History
The Electric Ray Torpedo torpedo
The “Birth” of Electrophysiology
 Animal Electricity=Bioelectronics

26th January
J 1781 L
Luigi
i i G
Galvani
l i
 Galvani’s Experiments

Galvani
G l i (1791) fi
first showed
h d that
h f frog muscle
l contracts whenh the
h muscle l or the
h iinnervating
i nerve iis
touched with a metal arch. Galvani interpreted his observation in analogy to the electric organ as a
discharge of electrical energy stored in the muscle cells.
 Dispute with Volta

http://www.answers.com/topic/voltaic-pile
 “Animal(=Bio) Electricity”

"Le docteur Ure galvanisant le corps de l'assassin Clydsdale."

An engraving from Les merveilles de la Science (1867) by Louis Figuier
Bioelectricity
 Let’s See Who was Right!

Cu2+ + 2e− → Cu (E = +0.34 V)

Zn2+ + 2e− → Zn (E = −0.76 V)
Fe (E= -044 V)

Thus the reaction that is going on is really:

Cu2+ + Zn → Cu + Zn2+

Th electric
The l potentiall is then
h +0.34
0 34 V −(−0.76
( 0 6 V) = 11.10
10 V

http://www.answers.com/topic/galvanic-cell
 Conductivity of Tissue

M1 – connective tissue, buffer zone between various

tissues - ρ1 = 10 Ωm;

M2 – tendons - ρ2 = 5 Ωm;

M3 – extracellular space
p - ρ3 = 6 Ωm;

M4 – blood vessels - ρ4 = 2.5 Ωm;

M5 – nerves - ρ5x = ρ5y = 10 Ωm ; ρ5z = 1 Ωm;

M6 – bone - ρ6 = 160 Ωm;

g - ρ7 = 40 Ωm;
M7 – cartilage

M8 – skin and fat - ρ8 = 20 Ωm;

M9 – articular
u disc - ρ9 = 60 Ωm;

M10 – muscles - ρ10x = ρ10y = 13.2 Ωm; ρ10z = 1.9 Ωm;

weight of adult frog: 173 ± 12.5 g;
mean leg weight:
weight 38 ± 5.1 g;
mean leg length : 16.8 ± 3.2 cm
http://cat.inist.fr/?aModele=afficheN&cpsidt=13935485
Under typical cellular conditions, ΔG of ATP is approximately −57 kJ/mol (−14 kcal/mol).
The ATP concentration inside the cell is typically 1 - 10 mM.

http://en.wikipedia.org/wiki/Adenosine_triphosphate
Basics of Memory
 Examples for Electrodes in Bio-Medicine
Deep Brain Pacemakers
Electrodes

Microelectrode
Arrays Glucose Sensor
What Makes the Monkey Smile?
 What Happens on the Electrode?

Ion binding

Electrolysis Potential

pH change
RE
Ionic
I i
strength
change
 Understanding the Bio-(Electronic)
Interface at the Nanoscale

Y Y Y
Surface

Bulk

Controlling
g the Function
 Useful Sources

Course material:
http://www.lbb.ethz.ch/Education/Biosensors

On the Web:

http://www corrosion-doctors
http://www.corrosion doctors.org/Biographies/GalvaniBio.htm
org/Biographies/GalvaniBio htm

http://www.answers.com/topic/galvanic-cell

http://en.wikipedia.org/wiki/Ion
http //en.wikipedia.org/wiki/Ion_gradient
gradient

http://www.informaworld.com/smpp/content~db=all~content=a751087996~tab=content
 Program 2010
22-Feb Introduction/Overview of the field and applications (Ekins paper, Galvani) Janos

01-March
01 March Basic notions of molecular adsorption and electron transfer Tomaso

08-March Potentiometric biosensors TZ

15-March Amperometric biosensors TZ
22 M
22-March h Opti l S
Optical Sensors
s s (OWLS Th Theory)) JV
29-March AFM for biosensing TZ
12-April Basics of electrophysiology Peter
Niederer
19-April Ion channels and patch clamp TZ
26-April Synapse and neuromuscular junction TZ
03-May Protein and DNA high throughput screening methods: Microarrays and JV
electrophoresis
l t h i (Fundamentals)
(F d t l )

10-May Acoustic Sensors (QCM theory) JV

17-May NanoBiosensors (Nanowire Sensors by Thomas Helbling, Plasmons by Takumi JV
S
Sannomiya)
i )

31-May Outlook, philosophy and risks (Related reading), Research highlights and JV
brain machine interface

HOMEWORK Exercises (2007 Excercises, 2007 Exam Questions) JV, TZ

 L2: Molecular Adsorption and Electron Transfer

Transition State Theory

A + B → [AB] → C
activated transition
complex state k
(precursor)
Gibbss energy G

activation
ΔG energy
(barrier)

reactants ΔG0 overall released

energy
products

reaction coordinate q

ΔG
Electron Transfer through a
kB
k∝e RT R=
NA
= 8.3 J ⋅ K −1 ⋅ mol−1
25
Molecular Wire

k el (x) = k 0ele−βx
β

Φ, barrier
4π 2mΦ in vacuum
β≈
h
 −1eV −1/2 Φ
≈ 1.02Α

34
 L3: Potentiometric Biosensors

Chemical Potential
dU = dq + dw (1st law of thermodynamics)

dq dw
Atkins

G = U + pV - TS (definition of Gibbs free energy)

 ∂U   ∂G 
μj ≡   μj ≡  
 ∂n   ∂n 
 j S,V,n'  j  p,T, n'
4 pH Electrode
thin glass
thi l membrane
b hi
highly l ti tto H+
hl selective
22% Na2O, 6% CaO, 72% SiO2
V
REind SiO − Na + + H + SiO − H + + Na +
REref
ppH ≡ −loga
g H+
ΔV = K'+0.059loga H + (V )
K'−ΔV
pH =
0.059

solution to be tested
16
► ion-selective electrodes (ISE)
 L4: Amperometric Biosensors
glucose
anode
2Fe(Cp)2 GODOx glucose

2e-

2Fe+(Cp)2 GODRed gluconolactone + 2H+

glucose + GODox → gluconolactone + GOD R + 2H +

GODR + 2Fe+ → GODox + 2Fe
2Fe - 2e- → 2Fe+
Three Electrodes Cell
Three-Electrodes
22
two - electrodes → E appl = E RE −sol + IR − E WE−sol

both depending on I
ee –electrodes
tthree e ec odes
cell E WE −sol too → bad

U RE U
= − in
R2 R1 U=0
U out
I=
R3

U=0

U WE = 0

27
 L5: Optical Sensors
Newton’s Optics 1704

n2

n1

Diffraction and Interference

Cover
C A

Waveguide θ F
Support S
α

lλ
N := nF sinθ = nair sinα +
Λ

2Π m = ΦF + ΦFS + ΦFAC
 L6: AFM for Biosensing
AFM Invention

FIG. I . Description of the principle operation of G. Binnig et al., Phys. Rev. Lett. 56 (1986) 530
an AFM. The tip follows contour B, to maintain
constant force between tip and sample (AFM,
sample, and tip either insulating or conducting).

problem of the tip approach to the substrate → how to avoid the crash of the tip!
3

AFM & Molecular Recognition

approach pulling away

(adhesion)

(jump to ccontact)
recognition
problem → how many molecules of the tip bind with the complementary
ones of the surface? Only one couple is formed or more? 17
L7: Basics of Electrophysiology
(Prof em.
(Prof. em Peter Niederer)

Model for propagation of

action potentiall
 L8: Ion channels and patch clamp
Ion Channels

Differentt ffrom aqueous pores

Diff
↳ ion selectivity
↳ not continuously open → gated 9
Patch-Clamp Recording

< 1 μm

12
E. Neher and B. Sakmann, 1991 Nobel Prize in Medicine
 L9: Synapse and neuromuscular junction

Electrical Synapse
Each gap junction (aka nexus junction)
contains numerous gap junction
channels which cross the membranes
of both cells. With a lumen diameter of
about 1.2 to 2.0 nm, the pore of a gap
junction channel is wide enough to
allow ions and even medium sized
molecules like signaling molecules to
flow from one cell to the next, thereby
connecting the two cells' cytoplasm.
Thus when the voltage of one cell
changes, ions may move through from
one cell to the next, carrying positive
charge with them and depolarizing the
postsynaptic cell.
from “wikipedia”

Gap junction channels are composed of two hemi-channels called

connexons in vertebrates, one contributed by each cell at the synapse.
Connexons are formed by six 7.5 nm long, four-pass membrane-
spanning protein subunits called connexins, which may be identical or
slightly different from one another. 10
Chemical Synapse
When an action potential reaches the nerve
terminal in a presynaptic cell, it stimulates
the terminal to release its neurotransmitter.
The neurotransmitter molecules are
contained in synaptic vesicles and are
released to the cell exterior when the
vesicles fuse with the plasma membrane of
th nerve tterminal.
the i l Th
The released
l d
neurotransmitter binds to and opens the
transmitter-gated ion channels concentrated
in the plasma membrane of the postsynaptic
target cell at the synapse. The resulting ion
flows alter the membrane potential of the
target cell, thereby transmitting a signal from
the excited nerve
nerve.

11
L10: Microarrays and electrophoresis
Microarray Readers

www zeptosens com

www.zeptosens.com

Sequencing with Gel Electrophoresis

Traditional Capillary
 L11: Acoustic Sensors
The QCM-D Technique

Decay-based QCM readout DNA hybridization

Larsson, L. et al., Anal Chem, 2003, 75:5080-5087.

QCM-D, optical readout and AFM

Rodahl M. et al., Rev Sci Instr, 1995, 66, 3924.

Höök F
Höök, F. et al
al., PNAS,
PNAS 1998,
1998 14,
14 1729
Reimhult et al., Anal Chem, 2004, 76, 7211
 L12: Nanobiosensors
Nanobiosensors: Ion-Channel Based Sensing Nanobiosensors: Nanowire Arrays

Nat. Biotechnol. 23, 1294-1301 (2005)

B.A. Cornell et al. Nature 387, 580
580-583
583 (1997)

Nanobiosensors: Transmission Plasmon Biosensor Immobilization of Single Vesicles on Nanopatterns

Au Ag

100nm 100nm

Angewandte Chemie International Edition 42, (45), 5580-5583, 2003

 L13: Brain Machine Interfaces
Examples for Electrodes in Bio-Medicine

Deep Brain Pacemakers

Electrodes

Microelectrode Arrays
Glucose Sensor
 Thank You!

Janos (Vörös),
(Vörös) Tomaso (Zambelli)

Laboratory of Biosensors and Bioelectronics

Institute for Biomedicall Engineering,
E

voros@ethz ch zambelli@ethz.ch
voros@ethz.ch, zambelli@ethz ch

http://lbb.ethz.ch/

Phone:
Janos: +41 44 632 59 03
Tomaso: +41 44 632 4575