Transducers in Biosensors I: Types and Principles

Transducers in Biosensors I
(Week 4)
bansi.malhotra@gmail.com
Centre for NanoBioengineering & Spintronics,

Chungnam National University,Daejeon,Korea
Transducers in Biosensors I
Various types of transducers: Principles and

applications
• Potentiometric/amperometric
conductrometric/resistormetric
• Optical
Calorimetric
Miscellaneous
10/5/2009 WCU Project,CNU,bansi.malhotra@gmail.com 2

1. Brief over view of Transducers
Transducer: a device that converts one form of
energy to another
• Energy forms as input and output
– Thermal: temperature, heat, heat flow,...
– Mechanical: position, velocity, acceleration,
force, pressure, ...
– Chemical: concentration, composition, reaction
rate…
– Optical: intensity, wavelength, phase, polarization
– Magnetic: field intensity, flux, magnetization, etc.
– Electrical: voltage, current, charge, …
Many measuring and sensing devices, as well as loudspeakers, thermocouples, microphones, and phonograph
pickups, may be termed transducers.
10/5/2009 WCU Project, CNU,bansi.malhotra@gmail.com 3

Transducer
IUPAC Definition :
• Transduction of the biosensor signal is a process
that is concurrent,and within the special
environment of the biosensing element.
• The transduction efficiency determines many of

the analytical characteristics of the biosensors,
such as the signal stability, reproducibility,
detection limit, and in many cases the operational
stability and selectivity.
Transducers in biosensors
The transducer converts the biochemical interactions into
measurable electronic signals. Electrochemical, electro-
optical, acoustical, and mechanical transducers are among
the many types used in biosensors. The transducer works
either directly or indirectly.
10/5/2009 Advances in Biosensors, B.D. Malhotra & A. P. F. Turner 5

Rasoolyet al, Biosensors , 37, 1 (2005)
WCU Project,CNU,bansi.malhotra@gmail.com
Responsivity and Detectivity
Responsivity = Output signal / Input signal
( Transducing effciency : if signals have same units )
Detectivity = Signal/Noise of output signal/

size of output signal
= Responsivity/output noise
signal
Sensor and Actuator
Sensor and actuator are kinds of transducers
Sensors:
Magnify/Demagnify an environmental perturbation
(signal/noise) and transform into an observable
energy form
Actuators:
Use small controlled energy to cause an
observable (or controllable) perturbation
(movement/energy radiation) to the environment

Types of Transducers commonly
used in biosensors
Transducers Examples
Enzymes,
Electrochemical Clark oxygen electrode, mediated electrode immunological
systems
1.Amperometric systems
Redox electrodes, ion selective electrodes, Gases, enzyme,
2.
Potentiometric field effect transistors, light addressable organelle, cell or
potentiometric sensors tissue;
electrodesIons in
3.Conductometri Platinum
measurementor gold electrodes for the biological media,
c the solution due to the generation of ionsof
of change in conductivity enzyme electrodes
pH; enzymes;
Optical Photodiode, waveguide systems, intergrate
optical sensors immunological
analytes
Acoustic (mass) Piezoelectric

devices, crystals, surface acoustic Volatile gases and
vapors, antibodies
Enzyme,
Thermistor or thermopile organelle, gases,
Calorimetric pollutants,
10/5/2009
antibiotics, 9
vitamins
1. Electrochemical sensor
Principle: Electrochemical reactions take place at electrode-
electrolyte interfaces and provide a switch for electricity to flow between
two phases of different conductivity, i.e. the electrode (electrons or holes
are the charge carriers) and solid or liquid electrolyte (ions are the main
charge carriers)
There are three types:

1) Potentiometric 2) Amperometric and 3) Conductometric. e.g Oxidation of Fe(CN)6
10/5/2009 Advances in Biosensors, B.D. Malhotra & A. P. F. Turner

1. AMPEROMETRIC BIOSENSORS
• With amperometric sensors, the electrode

potential is maintained at a constant level
sufficient for oxidation or reduction of the
species of interest (or a substance
electrochemically coupled to it).
• The current that flows is proportional to

the analyte concentration.
Id = nFADsC/d

AMPEROMETRIC BIOSENSORS
• Amperometric enzyme electrodes based

on oxidases in combination with hydrogen
peroxide indicating electrodes have
become most common among biosensors.
• With these reactions, the consumption of

oxygen or the production of hydrogen
peroxide may be monitored.
• The first biosensor developed was based

on the use of an oxygen electrode.
Typical Design
Auxiliary Electrode
(e.g. Pt wire)
Working Electrode Reference Electrode
(e.g. Pt, Au, C) (e.g. Ag/AgCl, SCE)
Buffer solution
(e.g. Tris, DPBS, Citrate)
e flow incorporating electrolyte
Stirbar (e.g. KCl, NaCl)
Example
Glucose
Glucose + O2
Oxidase
Gluconic Acid + H2O2
The product, H2O2, is oxidised at +650mV vs a

Ag/AgCl reference electrode.
Thus, a potential of + 650mV is applied and

the oxidation of H2O2 measured. This current
is directly proportional to the concentration
of glucose.

Typical curve
Typical Spectra
I (nA)
150
100
50
0
5 10 15 20
10/5/2009 [Glucose], mM 15
Clark Oxygen Electrode
-
+
Electrode body
Silver anode
KCl soln.
Polyethylene membrane
Platinum cathode
• The drawback of oxygen sensors is that they are
very prone to interferences from exogenous
oxygen.
• H2O2 is more commonly monitored. It is oxidised

at +650mV vs. a Ag/AgCl reference electrode.
• At the applied potential of anodic H2O2 oxidation,

however, various organic compounds (e.g.
ascorbic acid, uric acid, glutathione,
acetaminophen ...) are co-oxidised.
10/5/2009 17
• Various approaches have been taken to

increase the selectivity of the detecting
electrode by chemically modifying it by the
use of:
– Membranes
– Mediators
– Metallised electrodes
– Polymers

1. Membranes.
Various permselective membranes have been
developed which controlled species reaching the
electrode on the basis of charge and size.
Examples include cellulose acetate (charge and size),

Nafion (charge) and polycarbonate (size).
The disadvantage of using membranes is, however,

their effect on diffusion.

2. Mediators
Many oxidase enzymes can utilise artificial electron
acceptor molecules, called mediators.
A mediator is a low molecular weight redox couple

which can transfer electrons from the active site of
the enzyme to the surface of the electrode, thereby
establishing electrical contact between the two.
These mediators have a wide range of structures and

hence properties, including a range of redox potentials.

AMPEROMETRIC BIOSENSORS,
CV of FeCN

• Examples of commonly used mediators are:
– Ferrocene (insoluble)
– Ferrocene dicarboxylic acid (soluble)
– Dichloro-indophenol (DCIP)
– Tetramethylphenylenediamine (TMPD)
– Ferricyanide
– Ruthenium chloride
– Methylene Blue (MB)

3. Metallised electrodes
The purpose of using metallised electrodes is to create

conditions in which the oxidation of enzymatically
generated H2O2 can be achieved at a lower applied
potential, by creating a highly catalytic surface.
In addition to reducing the effect of interferents, due

to the lower applied potential, the signal-to-noise
ratio incresaes due to an increased electrochemically
active area.

Metallization is achieved by electrodepositing the
relevant noble metal onto a glassy carbon electrode
using cyclic voltammetry.
Successful results have been obtained from a few

noble metals - platinum, palladium, rhodium and
ruthenium being the most promising.

Glassy carbon electrode Metallised GCE
Potential
Potential
Glassy carbon electrodes do not catalyse the oxidation of

hydrogen peroxide.
GCEs metallised with ruthenium, rhodium, palladium or

platinum do.

4. Polymers
As with membranes, polymers are used to prevent

interfering species from reaching the electrode
surface. Polymers differentiate on the basis of size
and charge.
An example is that of polypyrrole. A polypyrrole

film has to be in the reduced state to become
permeable for anions. If the film is oxidised, no
anion can permeate.

• Examples of commonly used
polymers are:
– Polypyrrole
– Polythiophene
– Polyaniline
– Diaminobenzene
– Polyphenol

1.1 Amperometric transduction
Amperometry encompasses a group of electroanalytical
techniques. It monitors the current generated at a fixed
bias potential.
Linear relation between the concentration of the analyte

and the current generated is obtained.
Several techniques like cyclic voltammetery, flow

injection analysis studies, etc. are commonly employed.
Most biosensors are based on amperometric-type

detection.
Amperometric biosensors have been at the focus of electroanalytical

research since the first report of the enzyme electrode by Updike
and Hicks in 1967 for the detection of glucose.
Clark-type oxygen electrode
Example of Amperometric
sensor
Clark and Lyons 1962

1.2 Potentiometric transduction
A potentiometric biosensor monitors
the potential under zero current conditions. The potential
generated is directly proportional to the logarithm of the
analyte concentration.
The basis of this type of electrochemical monitoring is the
Potentiometric biosensors - these use ion-selective

electrodes to determine changes in concentration of
chosen ions, e.g., hydrogen ions.

POTENTIOMETRIC BIOSENSORS
• In potentiometric sensors, the zero-
current potential (relative to a reference)
developed at a selective membrane or
electrode surface in contact with a sample
solution is related to analyte
concentration.
• The main use of potentiometric
transducers in biosensors is as a pH
electrode.

• E = Eo + RT/nF ln[analyte]
– Eo is a constant for the system

– R is the universal gas constant
– T is the absolute temperature
– z is the charge number
– F is the Faraday number
– ln[analyte] is the natural logarithm of the
analyte activity.

• The best known potentiometric sensor is
the Ion Selective Electrode (ISE).
• Solvent polymeric membrane electrodes

are commercially available and routinely
used for the selective detection of several
ions such as K+, Na+, Ca2+, NH4+, H+, CO32-)
in complex biological matrices.
• The antibiotics nonactin and valinomycin

serve as neutral carriers for the
determination of NH4+ and K+, respectively.

Typical Cell
Ag/AgCl reference electrode
Internal aqueous
filling solution
Liquid ion exchanger
Membrane/salt bridge
10/5/2009 34
Porous membrane containing ionophore
• ISEs used in conjunction with immobilized
enzymes can serve as the basis of electrodes
that are selective for specific enzyme
substrates.
• The two main ones are for urea and

creatinine.
• These potentiometric enzyme electrodes are

produced by entrapment of the enzymes
urease and creatinase, on the surface of a
cation sensitive (NH4+) ISE.

+ urease + -
Urea + H2O + H 2NH4 + HCO3
creatininase +
Creatinine + H2O N-methylhydantoin + NH4
penicillinase
Penicillin Penicillonic Acid
In contact with pH electrode.

1.3 Conductometric transduction
Conductivity is a measurement of the ability of a solution to
conduct an
electric current. Instruments measure conductivity by placing
two plates of conductive material with known area and distance
apart in a sample. Then a voltage potential is applied and the
resulting current is measured. Using Ohms law
This is a technique where the changes
in ionic concentrations are measured. If the biocatalyst
produces
ionic products, or consumes ions, and the support
solution has
low electrical conductivity, this is often a
convenient and simple
technique.

Present status of electrochemical biosensors

Some electrochemical biosensors for
detection and estimation of analytes
10/5/2009 WCU 39
Project,CNU,bansi.malhotra@gmail.com
2. Optical Transducers
Principle: Optical sensors rely on the optical transduction

of the signal and comprise ultraviolet, visible and infrared
spectrophotometry in transmission or reflectance modes.
The relationship between the incident light intensity and
the transmitted radiation is given by the Lambert-Beer law

Optical Transducers
Optical methods have been used classically to monitor

analyte concentrations. Properties like absorption, refractive
indices, fluorescence, phosphorescence,
chemiluminescence, etc., can be used in order to monitor
the biological recognition in biosensors. The devices can be
miniaturized by using optical fibres, which act as light
guides. The detectors are often semiconductor photodiodes.
These devices are often used for remote analysis as the light
signal is resistant to electrical noise.

Examples of Optical Transducers
• Optical fibers
• Surface plasmon resonance sensors (SPR)
• Waveguide based SPR
• Integrated interferometers (Mach-Zehnder and Young
interferometers)
• Differential mode interferometry
• Resonant mirror
• Grating coupler
• Bidifractive couplers
• Optical waveguide lightmode spectroscopy system
(OWLS)
• Reflactometric interference spectroscopy (RIfS)

OPTICAL BIOSENSORS
• The area of biosensors using optical detection
has developed over the last number of years.
• The basis of these systems is that enzymatic

reactions alter the optical properties of some
substances allow them to emit light upon
illumination.
• Means of optical detection include fluorescence,

phosphorescence, chemi/bioluminescence...

OPTICAL BIOSENSORS
• Advantages of optical biosensors include
z due to fibre optics, miniaturisation is possible
z in situ measurements are possible
z in vivo measurements are possible
z diode arrays allow for multi-analyte detection
z signal is not prone to electromagnetic
interference
• Disadvantages include:
z ambient light is a strong interferent
z fibres are very expensive
z indicator phases may be washed out with time

OPTICAL BIOSENSORS
• Fibre optics are a sub-class of optical
waveguides which operate using the
principle of total internal reflection.
• Light incident on the interface between

two dielectric media is either reflected or
refracted according to Snell’s Law.

Working of SPR
The SPR is an optical phenomenon due to a charge density oscillation at
the interface of a metal and a dielectric, which have dielectric constants of
opposite signs
10/5/2009 WCU 46
Surface Plasmon Resonance
(i) Design: prisms coated with a thin film of metal, usually
gold or silver (~55-nm thick), typically known as the Kretschmann
configuration;by prism coupling: Working: a light wave passes
through a high-refractive-index prism and is totally reflected at the
prismϪmetal layer;
(ii) Design waveguide coupled with a thin film of metal;

Working by optical waveguide: the light wave is guided by an
optical waveguide and, when entering the region with a thin metal
layer, it evanescently penetrates through the metal layer exciting an
SPR at its outer boundary;
(iii) Design metallized diffraction gratings where the metal

thickness can be much greater (up to 150- nm thick)
Working: by light diffraction on a diffraction grating: the
component of the wave vector of the diffracted waves parallel to the
interface is diffraction-increased by an amount that is inversely
proportional
10/5/2009 to the period of the grating and can WCU be matched to that of 47
an SPR. Project,CNU,bansi.malhotra@gmail.com
Typical signal verses time curve
10/5/2009 WCU Project,CNU,bansi.malhotra@gmail.com ISBN: 3-540-4088-6

48
Typical Response Curve

Overview of optical biosensors

Comparison of sensitivities for
different optical biosensors
SPR has been one of the leading transducer techniques due to its extremely high sensitivity,
offering detection limits up to few ppt (pg mL−1 )

Bacteriorhodopsin:Biophotonic Material
Natural Transducer: Converts light into chemical energy
10/5/2009 52
Artificial Retina
4. Calorimetric
Principle
Calorimetric sensors are based on measurement of the heat
produced by
the molecular recognition reaction and the amount of heat
produced is cor-
related to the reactant concentration. Calorimetry can be
used for direct measurment of heat changes associated with
thermochemical proceses ( Grime, 1985).
The metabolic activity of the biocomponent causes an
increase in temperature, which is transformed into a
detectable electrical signal.
• Enzyme-catalysed reactions exhibit the
same enthalpy changes as spontaneous
chemical reactions.
• Considerable heat evolution is noted (5-
100kJ/mol).
• Thus, calorimetric transducers are 53
universally applicable in enzyme sensors.
Calorimetric Transducers
• The thermal biosensors constructed have

been based on:
z direct attachment of the immobilised enzyme or

cell to a thermistor
z Immobilisation of the enzyme in a column in

which the thermistor has been embedded.
ΔT = nΔH/cp

Typical Values of enthalpy
Enzyme Substrate -ΔΗ (kJ/mol)
Catalase Hydrogen peroxide 100.4
Cholesterol Cholesterol 52.9

oxidase
Glucose Glucose 80.0
oxidase
Hexokinase Glucose 27.6
Lactic Pyruvate 62.1

dehydrogenase
β - Lactamase Penicillin G 67.0
Urease Urea 6.6
Uricase Uric acid 49.1

10/5/2009 55
Bridge/Amplifier Recorder
Sample
Thermistor
Buffer
stream Enzyme reactor
Heat exchanger
Aluminium block polyurethane insulation
10/5/2009 56
10/5/2009 57
They are devices used to monitor the enthalpy change
of an enzyme-based reaction. If the enthalpy change
in the biocatalytic process is significant, then the
Thermistor temperature of the transducer (thermistor) is changed
and this change can be monitored. A thermistor is
nothing but a miniature resistance thermometer with
high sensitivity.
Two metals with different coefficient of expansion are

Bimetallic strip adjoined. On heating the entire strip bents toward one
end.
Liquid and gas
expansion
Metal-resistance
Pyroelectric
Thermal transducers
Radiant heat energy
sensing
Semiconductor-based Electrodes
These are transistor-like devices (usually npn type), and the

most common configuration is the field effect transistor
(FET). The biological element can be immobilized at the
surface of the gate of the FET to obtain EnFET (Enzyme
Field Effect Transistor). They operate in the potentiometric
Mode.

Liquid and gas expansion
Bimetallic strip
10/5/2009 WCU 60
Thermocouples
The EMF (in mV) at 100 C different for platinum/metal

thermocouples.

Metal-resistance sensors

Sound, infrasound and
ultrasound
Audio to electrical sensors and transducers
THE CARBON MICROPHONE
THE MOVING IRON (VARIABLE
RELUCTANCE) MICROPHONE
MOVING COIL MICROPHONE
RIBBON MICROPHONE
PIEZOELECTRIC MICROPHONES
CAPACITOR MICROPHONES
10/5/2009 WCU 63
Sound, infrasound and
ultrasound
Electrical to audio transducers
THE MOVING-IRON TRANSDUCER
THE MOVING-COIL TRANSDUCER
RIBBON LOUDSPEAKERS
PIEZOELECTRIC LOUDSPEAKERS
CAPACITOR TRANSDUCERS
ULTRASONIC TRANSDUCERS
INFRASOUND

Summary
• Transducer
• Types of Transducers : Electrochemical, Optical,
Thermal..
• Examples
• Next Week.....
10/5/2009 WCU 65
Transducers in Biosensors II(Week 5 )
Transducers Examples
Acoustic-Wave Microsensors, quartz
Piezoelectric crystal microbalance (QCM), Film bulk
acoustic wave resonators (FBARs)
LED, p-n diodes or bipolar junction
Semiconductor transducers, metal oxide semiconductor
(MOS) capacitor, field effect transistor
(FET)
Impedimetric
Mechanical Cantilevers
Molecular electronics Nanotransducers

based transducers
Key References
Advances in Biosensors: B. D. Malhotra and Anthany P. F.
Turner, Volume 5, 2003, Perspectives in biosensors
Enzyme and microbial biosensors: techniques and protocols ,
Ashok Mulchandani, Kim R. Rogers - 1998 - Science - 264
pages, Principles of Enzyme Biosensors, Ashok Mulchandani
& Kim Rogers, Humana Press
Sensor Technology Handbook, Jon S. Wilson - 2005 -
Technology & Engineering - 691 pages,6.5.Transduction
Mechanisms in Biosensors Conventional Transducers.
Mediated Biosensors,Asha Chaubey and B.D.Malhotra,
Biosensors & Bioelectronics, 2002, 17,Nos.6-7,pp441-456

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Transducers in Biosensors I

Centre for NanoBioengineering & Spintronics,

 Various types of transducers: Principles and

10/5/2009 WCU Project,CNU,bansi.malhotra@gmail.com 2

10/5/2009 WCU Project, CNU,bansi.malhotra@gmail.com 3

• The transduction efficiency determines many of

10/5/2009 Advances in Biosensors, B.D. Malhotra & A. P. F. Turner 5

Detectivity = Signal/Noise of output signal/

10/5/2009 WCU Project,CNU,bansi.malhotra@gmail.com 8

Acoustic (mass) Piezoelectric

There are three types:

10/5/2009 Advances in Biosensors, B.D. Malhotra & A. P. F. Turner

• With amperometric sensors, the electrode

• The current that flows is proportional to

10/5/2009 WCU Project,CNU,bansi.malhotra@gmail.com 11

• Amperometric enzyme electrodes based

• With these reactions, the consumption of

• The first biosensor developed was based

The product, H2O2, is oxidised at +650mV vs a

Thus, a potential of + 650mV is applied and

10/5/2009 WCU Project, CNU,bansi.malhotra@gmail.com 14

• H2O2 is more commonly monitored. It is oxidised

• At the applied potential of anodic H2O2 oxidation,

• Various approaches have been taken to

10/5/2009 WCU Project,CNU,bansi.malhotra@gmail.com 18

Examples include cellulose acetate (charge and size),

The disadvantage of using membranes is, however,

10/5/2009 WCU Project,CNU,bansi.malhotra@gmail.com 19

A mediator is a low molecular weight redox couple

These mediators have a wide range of structures and

10/5/2009 WCU Project,CNU,bansi.malhotra@gmail.com 20

10/5/2009 WCU Project,CNU,bansi.malhotra@gmail.com 21

• Examples of commonly used mediators are:

10/5/2009 WCU Project,CNU,bansi.malhotra@gmail.com 22

The purpose of using metallised electrodes is to create

In addition to reducing the effect of interferents, due

10/5/2009 WCU Project,CNU,bansi.malhotra@gmail.com 23

Successful results have been obtained from a few

10/5/2009 WCU Project,CNU,bansi.malhotra@gmail.com 24

Glassy carbon electrodes do not catalyse the oxidation of

GCEs metallised with ruthenium, rhodium, palladium or

10/5/2009 WCU Project,CNU,bansi.malhotra@gmail.com 25

As with membranes, polymers are used to prevent

An example is that of polypyrrole. A polypyrrole

10/5/2009 WCU Project,CNU,bansi.malhotra@gmail.com 26

10/5/2009 WCU Project,CNU,bansi.malhotra@gmail.com 27

Linear relation between the concentration of the analyte

Several techniques like cyclic voltammetery, flow

Most biosensors are based on amperometric-type

Amperometric biosensors have been at the focus of electroanalytical

Clark and Lyons 1962

Potentiometric biosensors - these use ion-selective

10/5/2009 WCU Project,CNU,bansi.malhotra@gmail.com 30

10/5/2009 WCU Project,CNU,bansi.malhotra@gmail.com 31

– Eo is a constant for the system

10/5/2009 WCU Project,CNU,bansi.malhotra@gmail.com 32

• Solvent polymeric membrane electrodes

• The antibiotics nonactin and valinomycin

10/5/2009 WCU Project,CNU,bansi.malhotra@gmail.com 33

Ag/AgCl reference electrode

Liquid ion exchanger

• The two main ones are for urea and

Various types of transducers: Principles and

THE MOVING IRON (VARIABLE

THE MOVING-COIL TRANSDUCER