Type of transducers used in biosensing

1. Electrochemical (Slides 6-29)

2. Optical (Slides 31-62)

3. Mechanical (Slides 63-67)

Electrochemical Transducers
Electrochemical transducers
In this the concentration of the ions generated in bio recognition
reaction are converted in to the

electrical voltage (Potentiometric),

electric current (Amperometric),

or electric resistance or conductance (Conductometric)

Cyclic voltammetry (Voltametric)

In this chapter, we will learn basic electrochemistry
An electrochemical cell is a device capable of either
1. Electrochemical cell generating electrical energy from chemical reactions or
using electrical energy to cause chemical reactions.

 The cells are constructed in separate

beakers.

 The metal electrodes are immersed in

electrolyte solutions.

 Each half-cell is connected by a salt

bridge, which allows for the free
transport of ionic species between the
two cells.
Reaction at the Reaction at the
anode:  When the circuit is complete, the
cathode: current flows and the cell “produces”
Zn(s) + 2e−→ Zn2+ Cu2+ + 2e−→ Cu(s) electrical energy.
Electrochemical cells generally consist of a cathode and an anode.

The key features of the cathode and the anode are tabulated below.

Anode Cathode

 Denoted by a negative sign since electrons are liberated here  Denoted by a positive sign since electrons are
consumed here
 An oxidation reaction occurs here
 A reduction reaction occurs in the cathode of an
 Electrons move out of the anode electrochemical cell

Reaction at the anode:  Electrons move into the cathode

Reaction at the cathode:
Zn(s) + 2e−→ Zn2+
Cu2+ + 2e−→ Cu(s)

Over all reaction:

Zn(s) + Cu2+→ Zn2+ + Cu(s)

 Electrolytic Cell
 Here, two inert electrodes are dipped
into molten sodium chloride (which
contains dissociated Na+ cations and
Cl– anions).
 When an electric current is passed
into the circuit, the cathode becomes
rich in electrons and develops a
negative charge.
•Reaction at  The positively charged sodium
•Reaction at cations are now attracted towards
Anode: 2Cl– Cathode: [Na+
+ e– → Na] x 2 the negatively charged cathode.
→ Cl2 + 2e–  This results in the formation of
metallic sodium at the cathode.
 Simultaneously, the chlorine atoms
•Cell Reaction: 2NaCl → 2Na + Cl2 are attracted to the positively
charged cathode.
An electrolytic cell can be defined as an electrochemical device that uses electrical  This results in the formation of
energy to facilitate a non-spontaneous redox reaction. chlorine gas (Cl2) at the anode (which
is accompanied by the liberation of 2
Electrolytic cells are electrochemical cells that can be used for the electrolysis of electrons, finishing the circuit).
certain compounds
In an electrolytic cell, the cathode is negative because the supplied

electrical energy to the cell results in the decomposition of chemical

compounds. The battery pumps electrons away from the anode

(making it positive) and into the cathode (making it negative).

What is Electrode?

Electrodes can be defined as conductors that are used to make electrical

contact with a non-metallic part of the circuit.
Types and Examples of Electrodes

Here we will look at some examples of electrodes.

There are mainly two types of electrodes, namely reactive and inert electrodes.

A reactive electrode is an electrode which actively participates in the reaction. Some reactive
electrodes include zinc, copper, lead, and silver.

An inert electrode is a type of electrode which does not participate in a chemical reaction.
Some commonly used inert electrodes include platinum, gold, graphite(carbon), and rhodium.
 Electrochemical measurements depend on the spontaneous interaction between electrical
energy and a chemical reaction involving an oxidation–reduction reaction to generate an
electrical current or vice versa.

 The chemical events that occur between immobilized biomaterials and the analytes result in
the production/consumption of ions or electrons, which has an effect on the electrical
current, the electrical potential, or any other electrical property of the solution.

 These reactions take place at the interface of a metal or semiconductor electrode and an
electrolyte

 Thus, detection is feasible if the reactions occur in close contact with the electrode surface.
Hence, the electrodes significantly influence the performance of the electrochemical
biosensor.

 One has to consider several factors when choosing a proper electrode, including its material,
dimension, and possibility to carry out surface modifications.
Most electrochemical cells are composed of
three electrodes:

• A reference electrode (RE): This electrode

is usually made of Ag/AgCl and stands at a
distance from the place where the reaction
occurs to provide a potential that is
proportional to the known and stable solution.
The RE allows normalizing of the
measurements.

• A counter (auxiliary) electrode (CE): This

electrode is the source of the current, which is
subsequently applied to the working electrode.

• A working electrode (WE) (the sensing or

redox electrode): This electrode acts as the
transduction element in the biochemical
reaction. CE and WE should be conductive and
chemically stable.
 Thus, depending on the analyte and the nature of the reaction, the mainly used electrode

materials are gold, silver, platinum, silicon, carbon, and graphene.

 An alternative to electrochemical cells is to screen-print the three electrodes on an insulating

substrate. These so-called screen-printed electrodes have several advantages, including

 simplicity,

 ease of mass production,

 low-cost construction,

 low analyte/reagent consumption

Potentiometric Biosensors
Potentiometric devices monitor the accumulation of a charge potential either at the WE compared
to the RE or between two REs separated by a semipermeable membrane.

Monitoring the potential occurs in an electrochemical cell under the condition that no significant
current flows between the two electrodes.

The transducer is usually an ion-selective electrode (ISE) composed of an electrochemical sensor

with a recognition element based on a thin film or a selective membrane.

The conversion of the biorecognition event into a potential signal by the ISE gives rise to a
measurable signal.

Example- Potentiometric enzyme biosensor for rapid determination of formaldehyde

based on succinimide-functionalized polyacrylate ion-selective membrane
One example of an enzyme electrode is the urea
electrode, which is based on the catalytic
hydrolysis of urea by urease

CO(NH2)2(aq)+ 2H2O(l) < = = = = > 2NH4+(aq)+

CO32–(aq)

The NH4+ that is produced is in equilibrium with

NH3

NH4+(aq)+H2O(l) < == == > H3O+(aq)+ NH3 (aq)

which, in turn, diffuses through the gas-permeable

membrane, where it is detected by a pH electrode.
Amperometric Biosensors
Amperometry means to measure the current that results from the electrochemical oxidation or
reduction of an electroactive species.

Applying a constant potential at the WE or on an array of electrodes with respect to the RE

induces the generated current, which is associated with the redox process.

In amperometric devices, the term “amperometry” refers to the technique characterized by

monitoring the current at a constant potential.
 Clark electrode At the anode metallic silver is oxidized to silver chloride:

KCl
At the cathode oxygen is reduced to water:

 The reaction chamber is separated from the electrodes by a

teflon membrane, which permits oxygen to diffuse from the
reaction buffer into the potassium chloride solution that
bathes the electrodes: a platinum cathode and a silver
anode.
 A voltage is applied between the electrodes and the
resulting current is proportional to the concentration of
oxygen.
 Typical Amperometric Glucose Biosensor

D-Glucose D-Glucono-1,5-lactone

 A potential is applied between the central platinum cathode and the annular silver
anode. This generates a current (I) which is carried between the electrodes by
means of a saturated solution of KCl. This electrode compartment is separated
from the biocatalyst (here shown glucose oxidase, GOD) by a thin plastic
membrane, permeable only to oxygen. The analyte solution is separated from the
biocatalyst by another membrane, permeable to the substrate(s) and product(s)

 The efficient reduction of oxygen at the surface of the cathode causes the oxygen
concentration there to be effectively zero.

 The rate of this electrochemical reduction therefore depends on the rate of diffusion
of the oxygen from the bulk solution, which is dependent on the concentration
gradient and hence the bulk oxygen concentration.
Ag anode 4Ag0 + 4Cl− 4AgCl + 4e−
 The reduction of oxygen concentration at cathode in directly proportional to the
amount of glucose concentration
Pt cathode O2 + 4H+ + 4e− 2H2O
Conductometric Biosensors
1. A conductometric biosensor is a type of biosensor that detects
biological molecules by measuring changes in electrical
conductivity.

2. The conductometric biosensors are based on the fact that almost all
enzymatic reactions involve either consumption or production of
charged species and, therefore, lead to a global change in the ionic
composition of the tested sample.

3. The conductivity of the electrolyte solution depends on the ion

concentration and mobility.

4. The conductometric transducer is a miniature two-electrode device

designed to measure the conductivity of the thin electrolyte layer
adjacent to the electrode surface.

5. Conductometric transducers are mostly manufactured by

microelectronics techniques – photolithography and vacuum
spraying.

6. These changes in conductivity are measured using conductometric

Conductometric biosensor techniques, typically by passing an alternating current (AC)
through the electrodes and measuring the resulting change in
conductivity. The change in conductivity is proportional to the
concentration of the analyte present.
7. Measurement and Detection: The generated electrical signal is measured and detected using appropriate electronic
instrumentation, such as conductance meters.

8. Reference electrode provide a stable electrical potential against which the conductance or conductivity changes can be
measured. Main role of the reference electrode is to maintain electrical balance and provide a consistent reference point for
conductivity measurements.

9. Reference signal refers to an alternating electrical signal generated by the instrument and used as a reference for phase-
sensitive detection. This reference signal serves as a phase reference against which the measured signal is compared. Phase-
sensitive detection, a technique employed by lock-in amplifiers to selectively extract and amplify the signal of interest
while rejecting noise and interference.

10. Biosensors based on the conductometric principle present a number of advantages: a) thin-film electrodes are suitable for
miniaturization and large scale production using inexpensive technology, b) transducers are not light sensitive, c) the driving
voltage can be sufficiently low to decrease significantly the power consumption, d) large spectrum of compounds of different
nature can be determined on the basis of various reactions and mechanisms.
Let's consider an example of a conductometric biosensor for glucose detection using an enzyme-
based recognition element, such as glucose oxidase (GOx).
Here's a step-by-step explanation of how it works:

Biological Recognition: Glucose oxidase (GOx) is immobilized onto the surface of a conductive transducer, such as a gold
electrode. The GOx enzyme selectively binds to glucose molecules present in the sample.

Sample Introduction: A sample containing glucose is introduced to the biosensor. Glucose molecules in the sample bind to the
GOx enzyme on the sensor surface.

Glucose Oxidation Reaction: When glucose binds to GOx, it undergoes an oxidation reaction, producing gluconic acid and
hydrogen peroxide (H2O2) as byproducts:

Glucose + O2 → Gluconic Acid + H2O2

Change in Conductivity: The presence of H2O2 produced in the oxidation reaction leads to a change in the electrical conductivity
of the transducer material. This change can be detected as a decrease in conductivity due to the consumption of electrons by the
oxidation reaction.

Transduction: The transducer converts the biochemical signal resulting from the glucose oxidation reaction into an electrical
signal that can be measured and analyzed. This transduction process may involve changes in electric conductivity.
Signal Amplification: The electrical signal generated by the transducer is typically weak and may be buried in noise. To improve
the signal-to-noise ratio (SNR), the signal may be amplified using electronic instrumentation, such as a lock-in amplifier. This
amplification process enhances the sensitivity and accuracy of the biosensor measurements.

Signal Detection: The amplified electrical signal is detected and measured using appropriate instrumentation, such as a
conductance meter or impedance analyzer. These instruments quantify the electrical response generated by the transducer,
providing information about the concentration of glucose in the sample.

Data Processing and Analysis: The measured electrical signal is processed, analyzed, and interpreted to determine the
concentration of glucose in the sample. This may involve calibration curves, standard reference materials, or mathematical
algorithms to correlate the signal response with glucose concentration.

Output: The final output of the biosensor provides a quantitative assessment of the glucose concentration in the sample. This
output may be displayed as a digital readout, graphical plot, or other format suitable for interpretation by the user.
Cyclic voltammetric Biosensors
Cyclic voltammetric biosensors operate on the principle of
utilizing cyclic voltammetry (CV) for the detection and
quantification of target analytes in biological samples.

Cyclic voltammetry is an electrochemical technique used to

study the redox properties of analytes in solution by
applying a potential sweep to the working electrode while
measuring the resulting current.

As the potential is swept back and forth, the redox reactions

occurring at the electrode surface generate characteristic
current peaks. The peaks in the cyclic voltammogram
correspond to the oxidation or reduction of the analyte at
specific potentials. By analyzing the peak shape, position, and
intensity, information about the concentration and kinetics of
the analyte can be extracted.

Cyclic voltammetric biosensors offer several advantages,

including high sensitivity, rapid response, and the ability to
perform real-time measurements in complex biological
samples.
Cyclic Voltammetric Glucose Biosensors

GOx-FADH2 (Reduced) H2O2

Glucose

Electrode
2e-
Glucolactone GOx-FAD (Oxidized) O2
Mediator

Cyclic-voltammogram of electrochemical
glucose sensor for a potential ranging from -0.8
V –0.8 V with a sweep rate of 100 mV/sec

Ref: 10.1557/adv.2016.149
Let's consider an example of a cyclic voltametric biosensor for glucose detection using an enzyme-based recognition
element, such as glucose oxidase (GOx).

Creating a cyclic voltammetric glucose biosensor involves several steps, each of which plays a crucial role in the functionality of
the biosensor. Below is a detailed step-by-step process, along with explanations of the role and working principle of each
component:

Step 1: Electrode Preparation

Role:
The electrode serves as the platform for the electrochemical reaction to occur and is essential for sensing glucose.

Working:
Choose an Electrode: Common choices include glassy carbon, platinum, or gold electrodes.
Clean the Electrode: Thoroughly clean the electrode surface to remove any contaminants that could interfere with the sensing
process.
Modify the Electrode: Depending on the specific design, the electrode may need modification to enhance its sensitivity and
selectivity towards glucose.

Step 2: Enzyme Immobilization

Role:
Immobilizing the enzyme (usually glucose oxidase, GOx) onto the electrode surface enables specific recognition and catalysis of
glucose. s, including medical diagnostics, food analysis, and bioprocess monitoring.
Working:
Prepare Enzyme Solution: Dissolve GOx in an appropriate buffer solution to maintain its stability and activity.
Immobilize GOx: Apply the enzyme solution onto the modified electrode surface, allowing it to adsorb or covalently attach. This
step can involve techniques like physical adsorption, cross-linking, or entrapment within a polymer matrix.

Step 3: Mediator Incorporation

Role:
The mediator facilitates electron transfer between the enzyme and the electrode, enhancing the electrochemical signal.

Working:
Choose a Mediator: Select a mediator with suitable redox properties, stability, and compatibility with the enzyme.
Prepare Mediator Solution: Dissolve the mediator in a suitable solvent or buffer.
Incorporate Mediator: Apply the mediator solution onto the enzyme-modified electrode surface, ensuring uniform coverage.
For example-

Glucose GOx-FADH2 (Reduced) Fe2+

Electrode
e-
Glucolactone GOx-FAD (Oxidized) Fe3+
Mediator
Step 4: Electrochemical Measurement
Role:
Cyclic voltammetry is employed to measure the electrochemical response of the biosensor and quantify glucose concentrations.

Working:
Setup Electrochemical Cell: Assemble the cell with the modified electrode as the working electrode, a reference electrode (e.g.,
Ag/AgCl), and a counter electrode (e.g., platinum).
Perform Cyclic Voltammetry: Apply a potential sweep to the working electrode while monitoring the resulting current. The
potential sweep should encompass the redox potentials of the mediator and the analyte (glucose).
Record Data: Collect cyclic voltammograms, which display characteristic peaks corresponding to the electrochemical reactions
involved in glucose oxidation and mediator regeneration.

Step 5: Calibration and Analysis

Role:
Calibration allows for the correlation of electrochemical signals with glucose concentrations in the sample.
Working:
Prepare Glucose Standards: Prepare a series of glucose solutions with known concentrations spanning the expected range.
Measure Cyclic Voltammograms: Analyze the cyclic voltammograms obtained from the glucose standards.
Construct Calibration Curve: Plot the peak current or peak potential against the corresponding glucose concentration to generate a
calibration curve.
Quantify Glucose Concentration: Use the calibration curve to determine the concentration of glucose in unknown samples based
on their electrochemical responses.
Step 6: Optimization and Validation
Role:
Optimization ensures the biosensor's performance meets the desired specifications, while validation confirms its reliability
and accuracy.
Working:
Optimize Parameters: Fine-tune experimental parameters such as enzyme loading, mediator concentration, and
measurement conditions to maximize sensitivity and minimize interference.
Validate Performance: Assess the biosensor's performance characteristics, including sensitivity, selectivity, reproducibility,
and stability, through rigorous testing with real samples.

By following these steps and understanding the role and working principles of each
component, you can develop a reliable cyclic voltammetric glucose biosensor for various
application
Optical Transducers
Optical
Transducers
• Light is a form of electromagnetic wave
that consists of oscillating electric and
magnetic fields.
• The wavelength of visible light ranges
from about 400 nm to 650 nm, which
defines several critical dimensions in optical
sensing techniques.

• The electric and magnetic fields of

propagating light are perpendicular to each
other and to the direction of propagation.
Such fields are oscillating harmonically in
temporal and spatial domains.

• Two electromagnetic waves can interfere,

which leads to an interference term that
contains a cosine of the phase difference
that can be positive or negative.
 The study of optics is a vital part of all sciences because it is
the basis of numerous technologies applied in the life
sciences, chemistry, physics, and engineering.

 More than 50 Nobel Laureates have contributed to optics

research, advancing studies on key technologies including
Raman scattering (awarded in 1930), laser (1950),
fluorescent proteins (2008), optical fibers (2009), and CCD
(2009).

 The advantages of optical transducers include lack of direct

contact, high spatial resolution, and relatively easy
detection.
 Optical transducers

Optical Surface plasmon Absorption Fluorescence

waveguides resonance Spectroscopy Spectroscopy
Optical waveguide
 An optical waveguide is a physical structure
that is designed to confine and guide light
waves along a specific path.
 These waveguides are used extensively in
various optical devices and systems, including
fiber optic communication networks,
optical sensors, and integrated photonic
circuits.
 The refractive index (often denoted as (n) of a material is defined as the ratio of
the speed of light in a vacuum (c) to the speed of light in the material (v).

Source : https://www.youtube.com/watch?v=kABd_fiRNm8&ab_channel=EMPossible
Source: https://www.youtube.com/watch?v=zA8UfR-Eekk&ab_channel=NajamAcademy
 The law of refraction

Where:

n1 is the refractive index of the initial medium (where the light is coming from)
n2​ is the refractive index of the medium the light is entering.
Θ1 is the angle of incidence, measured between the incident ray and the normal (a line
perpendicular to the interface) in the initial medium.
Θ2 is the angle of refraction, measured between the refracted ray and the normal in the
medium the light is entering.

Source : https://www.youtube.com/watch?v=kABd_fiRNm8&ab_channel=EMPossible
Source: https://www.youtube.com/watch?v=zA8UfR-Eekk&ab_channel=NajamAcademy
Source: https://www.youtube.com/watch?v=zA8UfR-Eekk&ab_channel=NajamAcademy
Source: https://www.youtube.com/watch?v=zA8UfR-Eekk&ab_channel=NajamAcademy
Key points about Snell's Law:

 It governs the bending of light as it passes from one medium to another with a different
refractive index.

 When light passes from a less dense to a more dense medium (e.g., air to glass), it bends
towards the normal. Conversely, when light passes from a more dense to a less dense
medium (e.g., glass to air), it bends away from the normal.

 If the angle of incidence is increased to the point where the angle of refraction is 90 degrees,
the light is said to be incident at the critical angle. Beyond this angle, total internal reflection
occurs.

 Snell's Law is applicable to various scenarios involving the refraction of light, including
lenses, prisms, and mirages, among others.
 Planar/slab
waveguide
• Slab waveguides are a waveguide with a planar
geometry. They are also called planar waveguides.
Planar waveguides consist of a core layer with a
higher refractive index surrounded by cladding
layers with lower refractive indices.
• When light enters the waveguide, it is confined
within the core due to the difference in refractive
index between the core and cladding.
• It guides light waves by total internal reflection.
• The top surface is usually used as the sensing site.
• The advantages of a planar structure include
easier fabrication and a larger sensing site
Rectangular
waveguides
• The structure of a rectangular
waveguide consists of four walls: two
parallel broad walls (top and bottom)
and two narrower sidewalls (sides).
• These walls are usually made of a
conductive material, such as metal,
which allows the waveguide to
confine and guide electromagnetic
waves.
• They are often used for
measurements that utilize
interferometry or optical resonation,
where the path lengths have to be
strictly confined to control
interference.
 Optical
fiber
• The most common type of optical waveguide
is the optical fiber, which consists of a core
surrounded by a cladding material with a
lower refractive index.
• Light is transmitted through the core by
repeatedly reflecting off the core-cladding
interface, allowing it to travel long distances
with low attenuation.
• The advantage of optical fibers for sensing
applications is their capability to be used as a
probe. Typically, one end of a fiber is used as
a sensing site. The fiber tip can be brought
into the sensing sites of in-situ or in-vivo
measuring applications.
Fiber Optic Core: the inner light-carrying
member with a high index of refraction.

Cladding: the middle layer, which serves to

confine the light to the core. It has a lower
index of refraction.

Buffer: the outer layer, which serves as a

"shock absorber" to protect the core and
cladding from damage. The coating usually
comprises one or more coats of a plastic
material to protect the fiber from the physical
environment. Sometimes metallic sheaths are
added to the coating for further physical
protection.
 Light injected into the fiber optic core and striking the core-to-cladding interface at an

angle greater than the critical angle is reflected back into the core.

 Since the angles of incidence and reflection are equal, the light ray continues to zigzag

down the length of the fiber.

 The light is trapped within the core. Light striking the interface at less than the critical

angle passes into the cladding and is lost.

Optical Waveguides notes: http://electron6.phys.utk.edu/optics421/modules/m8/Optical%20Wave%20Guides.htm

Total Internal Reflection: https://www.youtube.com/watch?v=Vcw3F32MSx4&ab_channel=Fliplearn.com

Optical fibers: https://www.youtube.com/watch?v=jZOg39v73c4&ab_channel=Lesics

Planar waveguide :
https://www.youtube.com/watch?v=kABd_fiRNm8&ab_channel=EMPossible

Rectangular waveguide :
https://www.youtube.com/watch?v=y4ReaugNtsk&ab_channel=EMPossible

http://electron6.phys.utk.edu/optics421/modules/m8/Optical%20Wave%20Guides.htm
Design of fiber
optic sensing
probe
Concanavalin A (Con A)-Glucose affinity optical biosensor
The lectin Concanavalin A (Con A) is one example of a protein that has binding sites for carbohydrates.

Fortunately, among the low-molecular-weight carbohydrate constituents present in blood plasma with the configurations necessary
to interact with Con A, only glucose occurs in significant concentrations.

Con A - FITC-dextran Con A — glucose FITC-dextran

glucose

Thus, given a method for the detection of the free FITC-dextran, one can estimate the glucose concentration.
The principle of competitive displacement is incorporated into a sensor device.
Here the Con A is immobilized on the inside wall of a dialysis hollow fiber.
By sealing one end of the hollow fiber and inserting a single optical fiber into the other end, a miniature chamber is formed that
comprises the transducer element of the sensor. The chamber is charged with FITC-dextran before sealing.
Exposure of the transducer to a solution containing glucose results in the diffusive penetration of glucose into the chamber.
Competition of glucose for Con A binding sites will displace FITC-dextran from the fiber wall and increase the free FITC-dextran
concentration.
The change in free dextran concentration is detected by measuring the fluorescence of the bulk solution by means of the optical
fiber and associated equipment, as shown in next Figure
JEROME S. SCHULTZ et al., 1982
Optical detection system in this biosensor
In this design, an optical fiber is used for transmitting the excitation light to the hollow fiber as
well as returning the fluorescent emission to the light detector. A single optical fiber has the
advantage of an efficient optical configuration; the field of view of the detector is the same as the
field of illumination, and a short path length is adequate for the collection of fluorescent light.

The optical system used here includes an argon laser as a source of excitation energy and
includes collector lens system, a dichroic beamsplitter (American Optical) to selectively reflect
and subsequently transmit desirable wavelengths, and a barrier filter (American Optical) to bar
unwanted wavelengths. The dichroic beamsplitter reflects the excitation light while longer
wavelengths, as emitted by fluorescence energy, pass through it and are not reflected from their
paths.
Surface plasmon resonance
A plasmon is defined as a quantum of collective electron oscillation in a solid.

Surface plasmon resonance (SPR) is a phenomenon that occurs where electrons in a thin metal
sheet become excited by light that is directed to the sheet with a particular angle of incidence,
and then travel parallel to the sheet.

 To excite surface plasmon polaritons in a resonant manner, one can use an incident light beam
(visible and infrared are typical). The incoming beam must match its momentum to that of the
plasmon.

Assuming a constant light source wavelength and that the metal sheet is thin, the angle of
incidence that triggers SPR is related to the refractive index of the material.

This makes SPR a possible technique for detecting substances (analytes) and SPR biosensors
have been developed to detect various important biomarkers.
SPR as transducer
SPR is an optical technique that relies on the
change in the refractive index of a metal surface
on which the target molecules are immobilized.

Upon incidence of light on a thin metal film at

a specific angle directed by a prism, it excites a
special electromagnetic wave known as surface
plasmon at the surface of the metal.

Because this incident angle is highly sensitive to

the dielectric environment on the opposite side
of the metal, any binding event on the metal
surface causes the angle to shift. This shift can
be measured non‐invasively in real time.

Source: https://www.reactionbiology.com/services/biophysical-assays/surface-plasmon-resonance-spr
 Absorbance Spectroscopy
According to the Beer-Lambert law, the
absorbance of a solution is directly
proportional to the concentration of the
absorbing material present in the solution and
path length.

The optical density or absorbance of a material

is a logarithmic intensity ratio of the light
falling upon the material, to the light
transmitted through the material.

The absorption spectrum is a very important

quantitative measure to evaluate optical
properties of solutions.

It provides several types of information such as

the concentration of a solute, shapes or sizes of
suspended particles, and biological activities.
The absorbance of a solution is usually
measured by preparing a sample in a cuvette or
a microwell plate.
Absorbance Spectroscopy as a transducer
Example
A colorimetric glucose sensor based on a conformational switch of i-motif DNA

Source: A colorimetric and ratiometric glucose sensor based on conformational switch of i-motif DNA,
https://doi.org/10.1016/j.talo.2020.100001
Reaction for this
biosensor
Working of A colorimetric glucose sensor based on a conformational switch of i-motif
DNA
 The i-motif DNA i-motifs are four-stranded DNA structures held together by hemi-
protonated and intercalated cytosine base pairs (C:C+)
 Therefore, in this reaction, first, the generation of gluconic acid in-situ supplied proton to the
solution through the glucose oxidized catalyzed reaction between glucose and O2, leading the
Poly(24C) to form i-motif structures.
 Because of this, MB that should have interacted with Poly(24C) was released into the
solution, inducing a color trend from purple (Poly(24C)-MB) to blue (free MB). More
glucose would liberate more MB from Poly(24C), which results in a more obvious color
change. Thus, a colorimetric and ratiometric glucose sensor could be realized.
Example of Fluorescence Biosensor: Concanavalin A (Con A)-Glucose affinity optical biosensor (Slide number 52-54)
Mechanical Transducers
Mechanical biosensors have
gained attention owing to their low cost,
sensitivity, ease of use, and rapid response.
The working principle of these types of
sensors relies on the transduction of
bio‐recognition interactions into a
mechanical signal. There are two main
types of mechanical biosensors:
1. cantilever‐based
2. piezoelectric
Principles of bio-sensing based on
microcantilever
  A cantilever beam is a structural element that is supported at one end and extends horizontally. The
unsupported end is called the cantilever and extends beyond the support point.

 Microcantilevers are ideal displacement sensors that can detect the motion of a cantilever beam with
nanometer precision.

 Cantilever bending can be related to the adsorption/desorption of molecules through adsorption forces.

 The working principle of nanomechanical transducers, and specifically of cantilevers, involves the
translation of the biochemical reaction occurring on top of the cantilever surface into a mechanical motion.
Any change produced on the sensing layer as a consequence of an external stimulus will cause a response
of the microcantilever.

(a) Cantilever biosensor (b) Bending of the cantilever due to

Source: https://doi.org/10.21203/rs.3.rs-3357343/v1 generated stress by interaction of analyte.
  Working in the static mode, the bending
arises as a consequence of a surface
stress change induced by any molecular
reaction that takes place on only one of
the cantilever surfaces.
 Stoney's equation is used to quantify the
cantilever displacement
surface stresses generated in molecular
thin films from different molecular
recognition events.
 In static mode, the change in cantilever
curvature is measured as a result of mass
change due to ligand-analyte interaction.
 The static mode needs a reference
electrode, and the bending of the
Stoney's equation cantilever as a result of biomolecular
adsorption can be mathematically
explained by Stoney's model as shown in
Figure
 Deflection Measurement: The deflection
of the cantilever is typically measured
using various techniques such as optical
interferometry, piezoresistive or
capacitive sensors, or atomic force
microscopy (AFM). These methods allow
for highly sensitive detection of even tiny
deflections induced by analyte binding.

Analyst, 2010, 135, 827–836

Fig. Schematic representation of the microcantilever-based biosensor working principles static mode
Optical based detection of microcantilever sensors
The displacement-based transducer
uses an optical detector that monitors
a reflected laser from the cantilever
surface, which changes its position as
a function of altered mass on it.

The optical method is simple to

implement and shows a linear
response with sub-angstrom
resolution and is currently the most
sensitive method for measuring
nanomechanical motions.

In the optical lever scheme, the

cantilever free end movement is
detected by measuring the reflected
laser beam displacement into a
Source: https://doi.org/10.1016/j.sintl.2020.100040 position-sensitive photodetector
(PSD).
Piezoelectric effect
 The piezoelectric effect refers to the phenomenon where certain materials generate an
electric charge in response to mechanical stress or deformation. Conversely, these materials
can also deform or change shape when subjected to an electric field.

This bidirectional coupling between mechanical and electrical properties occurs in

crystals with non-centrosymmetric structures, such as quartz, Rochelle salt, and certain
ceramics.

When a mechanical force is applied to a piezoelectric material, it causes the material's

lattice structure to distort, resulting in a redistribution of positive and negative charges
within the material, creating an electric potential across its surface.

This effect finds applications in various technologies, including sensors, actuators,

transducers, and energy harvesting devices.
Quartz Crystal Microbalance (QCM)
 The Quartz Crystal Microbalance (QCM) is a highly sensitive mass measuring device used
primarily in scientific research, material science, and various industries including chemical,
biological, and pharmaceutical.

 It is designed to detect minute changes in mass by measuring the change in resonance

frequency of a quartz crystal due to the deposition or removal of material on its surface.

https://www.researchgate
.net/publication/2423598 Quartz crystal microbalance
QCM gas sensor construction
55
Components of QCM:

Quartz Crystal: The heart of the QCM is a thin disk of quartz crystal with
precisely cut dimensions, typically in the range of a few millimeters in diameter and
a few hundred micrometers in thickness. Quartz is chosen for its stable and
predictable piezoelectric properties.

Electrodes: Thin metal electrodes are deposited on the surfaces of the quartz
crystal. These electrodes apply an alternating electric field to the crystal, inducing
its oscillation.

Electronics: QCM systems are equipped with electronic circuits to generate the
alternating voltage applied to the quartz crystal and to measure the resonant
frequency shift. Advanced QCM setups may include microprocessors for data
processing and analysis
Working of Quartz Crystal Microbalance (QCM)

The Quartz Crystal Microbalance (QCM) operates based on the principle of the piezoelectric
effect exhibited by quartz crystals.

When an alternating voltage is applied to the electrodes deposited on the surfaces of a thin
quartz crystal, it begins to oscillate at its resonant frequency.

As mass accumulates or dissipates on the crystal surface, induced by processes such as
biomolecular binding, or gas adsorption, the resonant frequency shifts proportionally to the
change in mass according to the Sauerbrey equation.

This frequency shift is detected and measured by electronic circuits, providing highly sensitive
and real-time mass measurements.

The QCM's ability to detect mass changes at the nanogram level makes it an invaluable tool in
various scientific and industrial applications, including thin film deposition, biosensing, and gas
sensing.
The Sauerbrey equation
Source:

https://www.youtube.com/watch?v=gXw7armpsEw&ab_channel=ZimmerPeacock

https://byjus.com/chemistry/electrochemical-cell/
https://byjus.com/chemistry/electrolytic-

cell/#:~:text=An%20electrolytic%20cell%20can%20be,the%20electrolysis%20of%20ce
rtain%20compounds.

https://byjus.com/chemistry/electrodes/

https://www.brainkart.com/article/Potentiometric-Biosensors---Potentiometric-
Methods-of-Analysis_29723/