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 • Optical

conductrometric/resistormetric

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

. • The transduction efficiency determines many of the analytical characteristics of the biosensors.Transducer IUPAC Definition : • Transduction of the biosensor signal is a process that is concurrent.and within the special environment of the biosensing element. and in many cases the operational stability and selectivity. reproducibility. detection limit. such as the signal stability.

acoustical.malhotra@gmail. 1 (2005) .bansi. Malhotra & A. Turner WCU Project.com 5 Rasoolyet al. B.CNU. 10/5/2009 Advances in Biosensors. P. Biosensors . The transducer works either directly or indirectly. F.D. 37. and mechanical transducers are among the many types used in biosensors. electrooptical. Electrochemical.Transducers in biosensors The transducer converts the biochemical interactions into measurable electronic signals.

10/5/2009 WCU Project.malhotra@gmail.CNU.com 6 .bansi.

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 10/5/2009 WCU Project. CNU.com 7 .bansi.malhotra@gmail.

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

Types of Transducers commonly used in biosensors Transducers Electrochemical 1. mediated electrode systems Redox electrodes. waveguide systems. ion selective electrodes. Calorimetric 10/5/2009 Thermistor or thermopile Enzyme. intergrate optical sensors pH. antibodies Acoustic (mass) Piezoelectric crystals. enzymes.Amperometric 2. antibiotics. Potentiometric 3. immunological systems Gases. light addressable potentiometric sensors Platinum or goldchange in conductivity of electrodes for the measurement of to the generation of ions the solution due Enzymes. electrodesIons in biological media. cell or tissue. enzyme electrodes Optical Photodiode. gases. organelle. pollutants. organelle.Conductometri c Examples Clark oxygen electrode. field effect transistors. 9 vitamins . immunological analytes Volatile gases and vapors. enzyme. surface acoustic devices.

B. Malhotra & A.CNU.D.bansi. P.1.e.malhotra@gmail. 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.g Oxidation of Fe(CN)6 10/5/2009 WCU Project. i. e.com Advances in Biosensors. Turner . F. Electrochemical sensor Principle: Electrochemical reactions take place at electrodeelectrolyte interfaces and provide a switch for electricity to flow between two phases of different conductivity.

1. AMPEROMETRIC BIOSENSORS • With amperometric sensors. • The current that flows is proportional to the analyte concentration.malhotra@gmail.bansi.com 11 .CNU. Id = nFADsC/d 10/5/2009 WCU Project. 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).

bansi. • The first biosensor developed was based on the use of an oxygen electrode.AMPEROMETRIC BIOSENSORS • Amperometric enzyme electrodes based on oxidases in combination with hydrogen peroxide indicating electrodes have become most common among biosensors. • With these reactions.malhotra@gmail. 10/5/2009 WCU Project. the consumption of oxygen or the production of hydrogen peroxide may be monitored.CNU.com 12 .

Citrate) incorporating electrolyte (e. Tris.com Stirbar Buffer solution (e.CNU.g. SCE) e flow 10/5/2009 WCU Project. NaCl) 13 . Pt.bansi.malhotra@gmail.g.Typical Design Auxiliary Electrode (e.g.g. Ag/AgCl. Pt wire) Working Electrode (e. Au. C) Reference Electrode (e.g. DPBS. KCl.

malhotra@gmail. 10/5/2009 WCU Project.bansi. a potential of + 650mV is applied and the oxidation of H2O2 measured. Thus.Example Glucose + O2 Glucose Oxidase Gluconic Acid + H2O2 The product. H2O2.com 14 . CNU. is oxidised at +650mV vs a Ag/AgCl reference electrode. This current is directly proportional to the concentration of glucose.

Typical curve Typical Spectra I (nA) 150 100 50 0 5 10/5/2009 10 15 20 15 [Glucose]. mM .

CNU.bansi.com 16 .malhotra@gmail. Polyethylene membrane Platinum cathode 10/5/2009 WCU Project.Clark Oxygen Electrode + Electrode body Silver anode KCl soln.

10/5/2009 17 .. uric acid. glutathione. however. a Ag/AgCl reference electrode. acetaminophen .) are co-oxidised. • H2O2 is more commonly monitored. It is oxidised at +650mV vs. ascorbic acid. various organic compounds (e.AMPEROMETRIC BIOSENSORS • The drawback of oxygen sensors is that they are very prone to interferences from exogenous oxygen.g.. • At the applied potential of anodic H2O2 oxidation.

AMPEROMETRIC BIOSENSORS • 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 10/5/2009 WCU Project.bansi.malhotra@gmail.com 18 .CNU.

Nafion (charge) and polycarbonate (size).AMPEROMETRIC BIOSENSORS 1. Various permselective membranes have been developed which controlled species reaching the electrode on the basis of charge and size. 10/5/2009 WCU Project.bansi. The disadvantage of using membranes is.malhotra@gmail. Examples include cellulose acetate (charge and size). however. their effect on diffusion.com 19 . Membranes.CNU.

AMPEROMETRIC BIOSENSORS 2. 10/5/2009 WCU Project.malhotra@gmail. 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. These mediators have a wide range of structures and hence properties. including a range of redox potentials. Mediators Many oxidase enzymes can utilise artificial electron acceptor molecules.CNU. called mediators.bansi. thereby establishing electrical contact between the two.com 20 .

com 21 .bansi.AMPEROMETRIC BIOSENSORS.malhotra@gmail. CV of FeCN 10/5/2009 WCU Project.CNU.

bansi.malhotra@gmail.CNU.com 22 .AMPEROMETRIC BIOSENSORS • Examples of commonly used mediators are: – Ferrocene (insoluble) – Ferrocene dicarboxylic acid (soluble) – Dichloro-indophenol (DCIP) – Tetramethylphenylenediamine (TMPD) – Ferricyanide – Ruthenium chloride – Methylene Blue (MB) 10/5/2009 WCU Project.

10/5/2009 WCU Project. the signal-to-noise ratio incresaes due to an increased electrochemically active area. In addition to reducing the effect of interferents.AMPEROMETRIC BIOSENSORS 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. due to the lower applied potential.malhotra@gmail.CNU.bansi.com 23 . by creating a highly catalytic surface.

com 24 .AMPEROMETRIC BIOSENSORS Metallization is achieved by electrodepositing the relevant noble metal onto a glassy carbon electrode using cyclic voltammetry. rhodium and ruthenium being the most promising. palladium.platinum.CNU. Successful results have been obtained from a few noble metals .malhotra@gmail.bansi. 10/5/2009 WCU Project.

10/5/2009 WCU Project. rhodium.Glassy carbon electrode Metallised GCE Potential Potential Glassy carbon electrodes do not catalyse the oxidation of hydrogen peroxide. palladium or platinum do.malhotra@gmail.CNU.com 25 .bansi. GCEs metallised with ruthenium.

polymers are used to prevent interfering species from reaching the electrode surface.com 26 . A polypyrrole film has to be in the reduced state to become permeable for anions. An example is that of polypyrrole. Polymers differentiate on the basis of size and charge. Polymers As with membranes. If the film is oxidised.malhotra@gmail.CNU.bansi. 10/5/2009 WCU Project.AMPEROMETRIC BIOSENSORS 4. no anion can permeate.

com 27 .bansi.CNU.AMPEROMETRIC BIOSENSORS • Examples of commonly used polymers are: – Polypyrrole – Polythiophene – Polyaniline – Diaminobenzene – Polyphenol 10/5/2009 WCU Project.malhotra@gmail.

malhotra@gmail. 10/5/2009 WCU Project. Linear relation between the concentration of the analyte and the current generated is obtained. It monitors the current generated at a fixed bias potential.CNU. Most biosensors are based on amperometric-type detection.bansi.1 Amperometric transduction Amperometry encompasses a group of electroanalytical techniques. flow injection analysis studies.com 28 . 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.1. etc. are commonly employed. Several techniques like cyclic voltammetery.

com 29 .malhotra@gmail.bansi.CNU.Clark-type oxygen electrode Example of Amperometric sensor Clark and Lyons 1962 10/5/2009 WCU Project.

bansi.com 30 .these use ion-selective electrodes to determine changes in concentration of chosen ions. 10/5/2009 WCU Project. hydrogen ions. e.1..malhotra@gmail.CNU. The basis of this type of electrochemical monitoring is the Potentiometric biosensors .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.g.

bansi. 10/5/2009 WCU Project.com 31 .malhotra@gmail. the zerocurrent potential (relative to a reference) developed at a selective membrane or electrode surface in contact with a sample solution is related to analyte concentration.CNU. • The main use of potentiometric transducers in biosensors is as a pH electrode.POTENTIOMETRIC BIOSENSORS • In potentiometric sensors.

10/5/2009 WCU Project.malhotra@gmail.com 32 .POTENTIOMETRIC BIOSENSORS • 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.CNU.bansi.

• Solvent polymeric membrane electrodes are commercially available and routinely used for the selective detection of several ions such as K+. respectively 10/5/2009 WCU Project.com 33 . H+.CNU. Na+.malhotra@gmail. determination of NH4 and K . NH4+. • The antibiotics nonactin and valinomycin serve as neutral carriers for the + + . Ca2+.bansi. CO32-) in complex biological matrices.POTENTIOMETRIC BIOSENSORS • The best known potentiometric sensor is the Ion Selective Electrode (ISE).

Typical Cell Ag/AgCl reference electrode Internal aqueous filling solution Liquid ion exchanger Membrane/salt bridge 10/5/2009 Porous membrane containing ionophore 34 .

POTENTIOMETRIC BIOSENSORS • ISEs used in conjunction with immobilized enzymes can serve as the basis of electrodes that are selective for specific enzyme substrates.bansi.CNU. • The two main ones are for urea and creatinine. • These potentiometric enzyme electrodes are produced by entrapment of the enzymes urease and creatinase. 10/5/2009 WCU Project.com 35 . on the surface of a + cation sensitive (NH4 ) ISE.malhotra@gmail.

POTENTIOMETRIC BIOSENSORS Urea + H2O + H Creatinine + H2O Penicillin + urease 2NH4 + HCO3 + - creatininase penicillinase N-methylhydantoin + NH4 Penicillonic Acid + In contact with pH electrode.malhotra@gmail.bansi. 10/5/2009 WCU Project.com 36 .CNU.

Using Ohms law This is a technique where the changes in ionic concentrations are measured.1.malhotra@gmail. Then a voltage potential is applied and the resulting current is measured.bansi. 10/5/2009 WCU Project.com 37 .3 Conductometric transduction Conductivity is a measurement of the ability of a solution to conduct an electric current.CNU. this is often a convenient and simple technique. and the support solution has low electrical conductivity. If the biocatalyst produces ionic products. or consumes ions. Instruments measure conductivity by placing two plates of conductive material with known area and distance apart in a sample.

bansi.com 38 .Present status of electrochemical biosensors 10/5/2009 WCU Project.CNU.malhotra@gmail.

malhotra@gmail.com 39 .Some electrochemical biosensors for detection and estimation of analytes 10/5/2009 WCU Project.bansi.CNU.

malhotra@gmail.2. visible and infrared spectrophotometry in transmission or reflectance modes.com 40 . Optical Transducers Principle: Optical sensors rely on the optical transduction of the signal and comprise ultraviolet.bansi. The relationship between the incident light intensity and the transmitted radiation is given by the Lambert-Beer law 10/5/2009 WCU Project.CNU.

These devices are often used for remote analysis as the light signal is resistant to electrical noise.com 41 . which act as light guides.CNU. The detectors are often semiconductor photodiodes. The devices can be miniaturized by using optical fibres.Optical Transducers Optical methods have been used classically to monitor analyte concentrations. phosphorescence.. 10/5/2009 WCU Project. fluorescence.malhotra@gmail. Properties like absorption. etc. chemiluminescence.bansi. can be used in order to monitor the biological recognition in biosensors. refractive indices.

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) 10/5/2009 WCU Project.bansi.com 42 .malhotra@gmail.CNU.

chemi/bioluminescence. • Means of optical detection include fluorescence.CNU.malhotra@gmail.. 10/5/2009 WCU Project.bansi. • The basis of these systems is that enzymatic reactions alter the optical properties of some substances allow them to emit light upon illumination. phosphorescence..OPTICAL BIOSENSORS • The area of biosensors using optical detection has developed over the last number of years.com 43 .

miniaturisation is possible in situ measurements are possible in vivo measurements are possible diode arrays allow for multi-analyte detection signal is not prone to electromagnetic interference • Disadvantages include: ambient light is a strong interferent fibres are very expensive indicator phases may be washed out with time 10/5/2009 WCU Project.OPTICAL BIOSENSORS • Advantages of optical biosensors include due to fibre optics.CNU.malhotra@gmail.bansi.com 44 .

OPTICAL BIOSENSORS • Fibre optics are a sub-class of optical waveguides which operate using the principle of total internal reflection.com 45 . 10/5/2009 WCU Project.bansi. • Light incident on the interface between two dielectric media is either reflected or refracted according to Snell’s Law.malhotra@gmail.CNU.

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 Project.com 46 .bansi.CNU.malhotra@gmail.

bansi. typically known as the Kretschmann configuration. usually gold or silver (~55-nm thick). (iii) Design metallized diffraction gratings where the metal thickness can be much greater (up to 150. Working by optical waveguide: the light wave is guided by an optical waveguide and. it evanescently penetrates through the metal layer exciting an SPR at its outer boundary.CNU.Surface Plasmon Resonance (i) Design: prisms coated with a thin film of metal. 47 .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 to the period of the grating and can be matched to that of 10/5/2009 WCU Project. when entering the region with a thin metal layer.com an SPR. (ii) Design waveguide coupled with a thin film of metal.malhotra@gmail.by prism coupling: Working: a light wave passes through a high-refractive-index prism and is totally reflected at the prismϪmetal layer.

CNU.Typical signal verses time curve 10/5/2009 WCU Project.malhotra@gmail.com 48 ISBN: 3-540-4088-6 .bansi.

CNU.malhotra@gmail.com 49 .Typical Response Curve 10/5/2009 WCU Project.bansi.

malhotra@gmail.bansi.Overview of optical biosensors 10/5/2009 WCU Project.CNU.com 50 .

bansi.CNU.malhotra@gmail. offering detection limits up to few ppt (pg mL−1 ) 10/5/2009 WCU Project.Comparison of sensitivities for different optical biosensors SPR has been one of the leading transducer techniques due to its extremely high sensitivity.com 51 .

Bacteriorhodopsin:Biophotonic Material Natural Transducer: Converts light into chemical energy 10/5/2009 Artificial Retina 52 .

4. • Thus. • Enzyme-catalysed reactions exhibit the same enthalpy changes as spontaneous chemical reactions. Calorimetric Principle Calorimetric sensors are based on measurement of the heat produced by the molecular recognition reaction and the amount of heat produced is correlated to the reactant concentration. calorimetric transducers are universally applicable in enzyme sensors. 1985). The metabolic activity of the biocomponent causes an increase in temperature. 53 . which is transformed into a detectable electrical signal. • Considerable heat evolution is noted (5100kJ/mol). Calorimetry can be used for direct measurment of heat changes associated with thermochemical proceses ( Grime.

Calorimetric Transducers
• The thermal biosensors constructed have been based on:
direct attachment of the immobilised enzyme or cell to a thermistor Immobilisation of the enzyme in a column in which the thermistor has been embedded.

ΔT = nΔH/cp
10/5/2009 WCU Project,CNU,bansi.malhotra@gmail.com 54

Typical Values of enthalpy
Enzyme Catalase Cholesterol oxidase Glucose oxidase Hexokinase Lactic dehydrogenase Substrate Hydrogen peroxide Cholesterol Glucose Glucose Pyruvate Penicillin G Urea Uric acid
WCU Project,CNU,bansi.malhotra@gmail.com

-ΔΗ (kJ/mol)
100.4 52.9 80.0 27.6 62.1 67.0 6.6 49.1
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β - Lactamase
Urease
10/5/2009

Uricase

Bridge/Amplifier

Recorder Thermistor

Sample

Buffer stream

Enzyme reactor

Heat exchanger
10/5/2009

Aluminium block

polyurethane insulation
WCU Project,CNU,bansi.malhotra@gmail.com

56

10/5/2009 WCU Project.CNU.malhotra@gmail.bansi.com 57 .

Thermistor They are devices used to monitor the enthalpy change of an enzyme-based reaction.com 58 .CNU.bansi. On heating the entire strip bents toward one end. Two metals with different coefficient of expansion are adjoined. then the temperature of the transducer (thermistor) is changed and this change can be monitored. If the enthalpy change in the biocatalytic process is significant. Bimetallic strip Liquid and gas expansion Metal-resistance Pyroelectric Thermal transducers Radiant heat energy sensing 10/5/2009 WCU Project. A thermistor is nothing but a miniature resistance thermometer with high sensitivity.malhotra@gmail.

malhotra@gmail.CNU.Semiconductor-based Electrodes These are transistor-like devices (usually npn type). They operate in the potentiometric Mode. The biological element can be immobilized at the surface of the gate of the FET to obtain EnFET (Enzyme Field Effect Transistor). 10/5/2009 WCU Project.bansi.com 59 . and the most common configuration is the field effect transistor (FET).

Liquid and gas expansion Bimetallic strip 10/5/2009 WCU Project.com 60 .CNU.malhotra@gmail.bansi.

CNU.com 61 .malhotra@gmail.Thermocouples The EMF (in mV) at 100 C different for platinum/metal thermocouples. 10/5/2009 WCU Project.bansi.

Metal-resistance sensors 10/5/2009 WCU Project.bansi.malhotra@gmail.CNU.com 62 .

bansi.com 63 . 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 Project.malhotra@gmail.CNU.Sound.

malhotra@gmail.bansi.com 64 .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 10/5/2009 WCU Project.CNU.

.Summary • Transducer • Types of Transducers : Electrochemical. Optical... • Examples • Next Week.com 65 .CNU. Thermal. 10/5/2009 WCU Project.malhotra@gmail.bansi...

field effect transistor (FET) Piezoelectric Semiconductor Impedimetric Mechanical Molecular electronics based transducers 10/5/2009 Cantilevers Nanotransducers 66 WCU Project.malhotra@gmail.com . metal oxide semiconductor (MOS) capacitor.bansi.CNU. quartz crystal microbalance (QCM). Film bulk acoustic wave resonators (FBARs) LED. p-n diodes or bipolar junction transducers.Transducers in Biosensors II(Week 5 ) Transducers Examples Acoustic-Wave Microsensors.

malhotra@gmail.Science . Biosensors & Bioelectronics.1998 . F.6-7. Jon S. Malhotra and Anthany P.com 67 . Kim R. Perspectives in biosensors Enzyme and microbial biosensors: techniques and protocols .264 pages.2005 Technology & Engineering . 2003. Wilson .691 pages. 2002.5.pp441-456 10/5/2009 WCU Project.6.bansi. Volume 5.D. 17.Asha Chaubey and B.Nos. Turner. D. Ashok Mulchandani.CNU. Ashok Mulchandani & Kim Rogers.Key References Advances in Biosensors: B. Mediated Biosensors. Principles of Enzyme Biosensors. Rogers .Transduction Mechanisms in Biosensors Conventional Transducers.Malhotra. Humana Press Sensor Technology Handbook.

malhotra@gmail.CNU.? 10/5/2009 WCU Project.bansi.com 68 .

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