UNIT I BIOPOTENTIAL ELECTRODES

Origin of bio potential and its propagation, Electrode-electrolyte interface, electrode-skin

interface, half-cell potential, contact impedance, polarization effects of electrode - non
polarizable electrodes, Types of electrodes - surface, needle and micro electrodes and their
equivalent circuits, Recording problems - motion artifacts and measurements with two
electrodes.

1. ORIGIN OF BIO POTENTIAL AND ITS PROPAGATION

ORIGIN OF BIO POTENTIAL

• Bioelectric potentials are produced as a result of electrochemical activity of a criteria

class of cells known as excitable cells that are components of nerves, muscular and
glandular tissue.
• The origins of biopotentials can be traced to the electric activity at the cellular cell.
• The electric potential across a cell membrane is the result of different ionic
concentrations that exist inside and outside the cell.
• There are two types of biopotentials are given below,

1. Action Potential
2. Resting Potential
Resting potentials

• The diffusion and drift processes give rise to membrane potential.

• The various ions seek a balance between the inside and outside of the cell by diffusion
and drift.
• But the membrane of excitable cells, such as nerve and muscle cells, readily permits
the entry of potassium and chloride ions while it effectively blocks the entry of
sodium ions.
• Due to difference in the permeability of different ions, the concentration of sodium
ions inside the cell becomes much lower than the outside the cell.
• Since the sodium ions are positive, the outside of the cell is more positive than the
outside. Thus balance is not achieved.
• However, an equilibrium is reached with a potential difference across the membrane
such that negative on the inside and positive on the outside.
• This membrane potential caused by the different concentration of ions is called resting
potential of the cell

Characteristics of resting potential

• The value of resting potential is maintained as a constant until some kind of

disturbance upsets the equilibrium.
• It is strongly depending on temperature.
• Since the permeability of different cell types vary, the corresponding resting
potentials vary as well. Thus it varies from -60 to -100 mV.
 Figure - Polarized cell with its resting potential

Action potentials

• When a section of the cell membrane is excited by the flow of ionic current or by
some form of externally applied energy, the permeability of the membrane changes so
that the sodium ions are allowed to enter inside the cell.
• This movement of sodium ions into the cell constitutes an ionic current which further
reduces the barrier of the membrane to sodium ions.
• The net result is an avalanche effect such that sodium ions rush into the cell and try to
balance with the ions outside.
• Meanwhile potassium ions are leaving the cell but are unable to move as rapidly as
the sodium ions.
• Therefore the cell has a slightly positive potential on the inside due to the imbalance
of the potassium ions.
• This positive potential of the cell membrane during excitation is called action
potential and is about 20 mV.
• As long as the action potential exists, the cell is said to be depolarized.

Figure - Depolarized cell and Cell with action potentials

• When the passage of sodium ions is stopped, the ionic currents that lowered the
barrier to sodium ions are no longer present and the membrane reverts back to the
original (polarized) condition.
• By the action of the sodium pump, the sodium ions are quickly transported to the
outside of the cell and the cell is in its resting potential.
 • Generally in nerve and muscle cells repolarization occur so rapidly following
depolarization that the action potential appears as a spike of as little as 1 millisecond
total duration.
• But for heart muscle, the action potential is withstanding from 150 to 300
milliseconds and so it repolarizes much more slowly.

Figure - Waveform of the action potential

• It shows a typical action-potential waveform, beginning at the resting potential,

depolarizing, and returning to the resting potential after repolarization.

PROPAGATION OF ACTION POTENTIALS

Figure – Electrical Activity associated with one contraction in a muscle

• When a cell is excited and generates an action potential, ionic currents begin to flow.
This process can, in turn, excite neighbouring cells or adjacent areas of the same cell.
• The rate at which an action potential moves down a fiber or is propagated from cell to
cell is called the propagation rate. In nerve fibers the propagation rate is also called
the nerve conduction rate, or conduction velocity.
• The usual velocity range in nerves is from 20 to 140 meters per second (m/sec).
Propagation through heart muscle is slower, with an average rate from 0.2 to 0.4
m/sec.
• Special time-delay fibers between the atria and ventricles of the heart cause action
potentials to propagate at an even slower rate, 0.03 to 0.05 m/sec.

2. ELECTRODE-ELECTROLYTE INTERFACE

• The most commonly used electrodes in patient monitoring and related studies are
surface electrodes.
• In order to avoid movement artefacts and to obtain a clearly established contact (low
contact impedance) an electrolyte or electrode paste is usually employed as an
interface between the electrode and the surface of the source of the event.

Figure - Electrode-tissue interface for surface electrodes used with electrode jelly

• The characteristic of a surface electrode composed of a metal electrode and

attached to the surface of the body through an electrolyte (electrode jelly) are
dependent upon the conditions at the metal-electrolyte interface, the electrolyte-
skin interface and the quality of the electrolyte.
• At the electrode-electrolyte transition, there is a tendency for each electrode to
discharge ions into the solution and for ions in the electrolyte to combine with
each electrode.
 Figure - Electrode tissue interface

• Electrode tissue interface circuit involves transfer of electrons from the metal
phase to an ionic carrier in the electrolyte, a charge double layer (capacitance)
forms at the interface.
• The net result is the creation of a charge gradient (difference of potential) at each
electrode, the spatial arrangement of which is called the electrical double layer.
• The double layer is known to be present in the region immediately adjacent to the
electrode and can be represented, in its simplest form, as two parallel sheets of
charge of opposite sign separated by a thin film of dielectric.
• Therefore, the electrode - electrolyte interface appears to consist of a voltage
source in series with a parallel combination of a capacitance and reaction
resistance. The voltage developed is called the half-cell potential.

Figure - Charge distribution at electrode-electrolyte interface

• To a first-order approximation, the half-cell potential is equal to the electrode

potential of the electrode, if the electrodes were used in a chemical measuring
application.
• All electrode potentials are measured with respect to a reference electrode, usually
that of hydrogen absorbed on platinum black.
• Electrode potentials of some of the commonly used metals in the electrochemical
series with respect to Hydrogen.

• The net current flows from electrode region to electrolyte in any measuring
device likeECG, EEG, EMG and EOG etc.
• The current crosses it from left to right. The electrode consists of metallic atoms
C. The electrolyte is an aqueous solution containing cations of the electrode metal
C + and anions A - the current in the electrode.
• The net current that cross the interface, passing from electrode to the electrolyte
consistof three process:
1. Electrons (e-) moving in a direction opposite to that of current in
electrode
2. Cations (C+) moving in same direction as current.
3. Anions (A-) moving in a direction opposite to that of current in
electrolyte.
• For charge to cross the interface there are no free electrons in the electrolyte and
no free cations or anions in the electrode-something must occur at the interface
that transfers the charge between these carriers.
• What actually occur are chemical reactions at the interface, which can be
represented in general by the following reactions:
where n is the valence of C and m is the valence of A.

• Electrode is made up of some atoms of the same material as the cations and that this
material in the electrode at the interface can become oxidized to form a cation and one
or more free electrons. The cation is discharged into the electrolyte; the electron
remains as a charge carrier in the electrode.
• In this case an anion coming to the electrode-electrolyte interface can be oxidized to a
neutral atom, giving off one or more free electrons to the electrode.
• Both reactions are often reversible and that reduction reactions (going from right to
left in the equations) can occur as well.
• As a matter of fact, when no current is crossing the electrode-electrolyte interface,
these reactions often still occur.
• But the rate of oxidation reactions equals the rate of reduction reactions, so the net
transfer of charge across the interface is zero.
• When the current flow is from electrode to electrolyte, the oxidation reactions
dominate.
• When the current is in the opposite direction, the reduction reactions dominate.

3. ELECTRODE-SKIN INTERFACE

• When bio potentials are recorded from the surface of the skin, we must consider the
additional interface between the electrode-electrolyte and the skin.
• In coupling an electrode to the skin, we generally use transparent electrolyte gel
containing Cl- as the principal anion to maintain good contact.
• Alternatively, we may use an electrode cream, which contains Cl- and has the
consistency of hand lotion.
• The interface between this gel and the electrode is an electrode-electrolyte interface.
However, the interface between the electrolyte and the skin is different.
• The skin consists of three principal layers that surround the body to protect it from its
environment.
• The outermost layer, or epidermis, plays the most important role in the electrode-skin
interface.
• This layer, which consists of three sub layers, is constantly renewing itself.
• The electrode-electrolyte interface equivalent circuit is shown adjacent to the
electrode-gel interface.
• The series resistance Rs, is now the effective resistance associated with interface
effects of the gel between the electrode and the skin.
• The epidermis, or at least the stratum corneum, is a membrane that is semipermeable
to ions, so if there is a difference in ionic concentration across this membrane, there is
a potential difference Ese, which is given by the Nernst equation.

Figure - Equivalent circuit of electrode skin interface

• The epidermal layer has an electric impedance that behaves as a parallel RC circuit.
• For 1 cm², skin impedance reduces from approximately 200 kn at 1 Hz to 200 2 at 1
MHz.
• The dermis and the subcutaneous layer under it behave in general as pure resistances.
They generate negligible de potentials.
• If the effect of the stratum corneum can be reduced, a more stable electrode results.
• We minimize the effect of the stratum corneum by removing it, or at least a part of it,
from under the electrode.
• There are many ways to do this, ranging from vigorous rubbing with a pad soaked in
acetone to abrading the stratum corneum with sandpaper to puncture it.
• In all cases, this process tends to short out Ese, Ce, and Re, thereby improving the
stability of the signal, but the stratum corneum can regenerate in as short a time as 24
h.
• Galvanic skin reflex (GSR) is the contribution of the sweat glands and sweat ducts.
The fluid secreted by sweat glands contains Na+, K+, and Cl ions, the concentrations
of which differ from those in the extracellular fluid.
• Thus, there is a potential difference between the lumen of the sweat duct and the
dermis and subcutaneous layers. There also is a parallel RpCp combination in series
 with this potential that represents the wall of the sweat gland and duct, as shown by
the broken lines in Figure.
• These components are often neglected when we consider biopotential electrodes
unless the electrodes are used to measure the electrodermal response or GSR.

4. HALF-CELL POTENTIAL (OR) ELECTRODE POTENTIAL

• The voltage developed at an electrode electrolyte interface is designated as the 'Half-

cell potential' or 'electrode potential'. In the case of a metal-solution interface,
electrode potential results from the difference in rates between two opposing
processes:
1. the passage of ions from the metal into the solution and
2. the combination of metallic ions in solution with electrons in the metal to form
atoms of the metal.
• So when a metal electrode comes into contact with an electrolyte (body fluid), there is
a tendency for the electrode to discharge ions into solution and for ions in the
electrolyte to combine with the electrode.
• The net result is the creation of a charge gradient, the spatial arrangement of which is
called the electrical double layer. In practice, it is not a stable and its variations
constitute a source of variable noise voltage, called artifacts.

Figure - Surface electrode equivalent circuit

• Figure shows the electrical equivalent circuit of a surface electrode when it is in

contact with the body surface.
• The impedance of this equivalent circuit can be written as
 • The electrode-electrolyte interface resembles a voltage source having half-cell
potential 'E' which is developed due to charge gradient and a capacitor 'C in parallel
with a leakage resistance 'R.
• The series resistance in the equivalent circuit 'R represents the series electrolyte and
skin resistance under equilibrium conditions.

Where,

Ehc - half-cell potential

Cd- electrode capacitance leakage resistance
Rd – leakage resistance
Rs - series electrolyte and skin resistance

• The value of the voltage and impedance depend on the electrode metal, its area,
electrolyte, charge density and frequency of current.
• The half-cell potential or electrode is measured with reference to hydrogen electrode
placed in the electrolyte near that metallic electrode.
• The half-cell potential developed can be expressed by the Nernst equation:

Where,

R = gas constant = 8.314 kJ/k mol/K

T = absolute temperature in Kelvin
F = number of coulombs transferred (or) Faraday constant = 96500 columbs.
n - valency of the ion
C1, C2 = concentration of the selected ion on the two sides of the membrane.
f1f2 = activity coefficients of the ion on the two sides of the membrane.
In dilute solutions f₁ = f₂ = 1.

5. CONTACT IMPEDANCE

• The bioelectrical events are usually recorded by means of metallic electrodes placed
on the surface of the body.
• The electrical activity generated by various muscles and nerves within the body is
conducted to the electrode sites through the body tissues, reaches the electrodes
through the skin electrode transition and is then conducted by direct wire connection
to the input circuit of the recording machine.
• The impedance at the electrode-skin junction comes in the overall circuitry of the
recording machine and, therefore, has significant effect on the final record.
• Skin electrode impedance is known as the contact impedance and is of a value much
greater than the electrical impedance of the body tissue as measured beneath the skin.
 • The outer horny layer of the skin is responsible for the bulk of the skin contact
impedance and, therefore, a careful skin preparation is essential in order to obtain best
results.

6. POLARIZATION EFFECTS OF ELECTRODE

• The half-cell potential of an electrode is no electric current exists between the

electrode and the electrolyte.
• If, on the other hand, there is a current, the observed half-cell potential is often
altered.
• The difference is due to polarization of the electrode.
• The difference between the observed half-cell potential and the equilibrium zero-
current half- cell potential is known as the over potential.
• Three basic mechanisms contribute to this phenomenon, and the over potential can be
separated into three components: the ohmic, the concentration, and the activation
over potentials.

Figure – waveform of overpotential

Ohmic over potential

• The ohmic over potential is a direct result of the resistance of the electrolyte. When a
current passes between two electrodes immersed in an electrolyte, there is a voltage
drop along the path of the current in the electrolyte as a result of its resistance.
• This drop in voltage is proportional to the current and the resistivity of the electrolyte.
• The resistance between the electrodes can itself vary as a function of the current.
• Thus the ohmic over potential does not necessarily have to be linearly related to the
current. This is especially true in electrolytes having low concentrations of ions.
• This situation, then, does not necessarily follow Ohm's law.

Concentration over potential

• The concentration over potential results from changes in the distribution of ions in the
electrolyte in the vicinity of the electrode-electrolyte interface.
 • Recall that the equilibrium half-cell potential results from the distribution of ionic
concentration in the vicinity of the electrode-electrolyte interface when no current
flows between the electrode and the electrolyte.
• Under these conditions, reactions and reach equilibrium, so the rates of oxidation and
reduction at the interface are equal.
• When a current is established, this equality no longer exists. Thus it is reasonable to
expect the concentration of ions to change.
• This change results in a different half-cell potential at the electrode.
• The difference between this and the equilibrium half-cell potential is the
concentration over potential.

Activation over potential

• The third mechanism of polarization results in the activation over potential.

• The charge-transfer processes involved in the oxidation-reduction reaction are not
entirely reversible.
• In order for metal atoms to be oxidized to metal ions that are capable of going into
solution, the atoms must overcome an energy barrier.
• This barrier, or activation energy, governs the kinetics of the reaction.
• The reverse reaction-in which a cation is reduced, thereby plating out an atom of the
metal on the electrode-also involves an activation energy, but it does not necessarily
have to be the same as that required for the oxidation reaction.
• When there is a current between the electrode and the electrolyte, either oxidation or
reduction predominates, and hence the height of the energy barrier depends on the
direction of the current.
• This difference in energy appears as a difference in voltage between the electrode and
the electrolyte, which is known as the activation over potential.

These three mechanisms of polarization are additive. Thus the net over- potential of an
electrode is given by
 7. NON POLARIZABLE ELECTRODES

Perfectly Polarizable Electrodes

• Electrodes in which no actual charge crosses the electrode-electrolyte interface when

a current is applied.
• The current across the interface is a displacement current and the electrode behaves
like a capacitor.
• Overpotential is due concentration.
• Example: Platinum electrode

Perfectly Non-Polarizable Electrode

• Electrodes in which current passes freely across the electrode- electrolyte interface,
requiring no energy to make the transition.
• These electrodes see no overpotentials.
• Example: Ag/AgCl Electrode
• Example: Ag-AgCl is used in recording while Pt is used in stimulation

Ag/AgCl Electrode

• Approach the characteristic of a perfectly non polarizable electrode Advantage of

Ag/AgCl is that it is stable in liquid that has large quantity of Cl- such as the
biological fluid.
• Ag/AgCl exhibits less electric noise than the equivalent metallic Ag electrode.

• A silver metal base with attached insulated lead wire is coated with a layer of the
ionic compound AgCl.
• This material-AgCl-is only very slightly soluble in water, so it remains stable.
• The electrode is then immersed in an electrolyte bath in which the principal anion
of the electrolyte is Cl-.
 • For best results the electrolyte solution should also be saturated with AgCl so that
there is little chance for any of the surface film on the electrode to dissolve.

Figure – Ag/AgCl Electrode

• The second reaction occurs immediately after the formation of Ag ions. These
ions combine with Cl ions already in solution to form the ionic compound AgCl.
• As mentioned before, AgCl is only very slightly soluble in water, so most of it
precipitates out of solution onto the silver electrode and contributes to the silver
chloride deposit.
• Silver chloride's rate of precipitation and of returning to solution is a constant K,
known as the solubility product.
• The first and second terms on the right-hand side of are constants; only the third is
determined by ionic activity. In this case, it is the activity of the Cl- ion, which is
relatively large and not related to the oxidation of Ag which is caused by the
current through the electrode.
• The half-cell potential of this electrode is consequently quite stable when it is
placed in an electrolyte containing Cl as the principal anion, provided the activity
of the CI remains stable.

8. TYPES OF ELECTRODES - SURFACE, NEEDLE AND MICRO ELECTRODES

AND THEIR EQUIVALENT CIRCUITS

Electrode

• Bio potential electrodes are those which help in picking up the electrical signals of the
body.
• These electrical signals are a consequence of the chemical activity in the biological
system.
• Chemical activity is due to ions and this chemical activity takes place at cellular level.
Ions like Na+, Cl- and K+ are predominantly present.
• The concentration gradient and its unbalanced condition inside and outside the cell
lead to electrical activity, which is picked up by bio potential electrodes.
Types of electrodes

1. Microelectrodes: Electrodes used to measure bioelectric potentials near or within a single

cell.

2. Skin surface electrodes: Electrodes used to measure ECG, EEG, and EMG potentials
from the surface of the skin.

3. Needle electrodes: Electrodes used to penetrate the skin to record EEG potentials from a
local region of the brain or EMG potentials from a specific group of muscles.

1. Microelectrodes

• Microelectrodes are divided into metallic and non-metallic.

• Non-metallic microelectrode is called micro-pipet.
• The microelectrodes should have smaller diameter and during insertion of the
electrode into cell, there will not be any damage to the cells.
• When a microelectrode is used to measure the potential of the cell, it is located within
the cell, while the reference electrode is situated outside the cell.
• The size of the electrode is determined by the size of the cell.
• Since the size of the cells is about 50 microns, the diameter of the tip of the
microelectrodes is ranging from 0.5 to 5 microns.

Types of Surface electrodes

• Metal microelectrode
• Micropipette

Metal microelectrode

• Metal microelectrodes are formed by electrolytically etching the tip of a fine tungsten
or stainless steel wire to a fine point.
• This technique is known as electropointing.
 • The metal microelectrodes are coated almost to the micro tip with an insulating
material. To reduce the impedance, some electrolytic processing like chloriding the
tip and then developing by the photographic developer can be performed.

Figure (a) shows the position of the electrodes and figure (b) is the electrical equivalent of
figure (a). Since the measurement of bioelectric potentials requires two electrodes, the
voltage measured is really the difference between the instantaneous potentials of the
microelectrode and the reference (indifferent) electrode and it is the sum of three potentials as
shown in figure 2.3(b) such that

• EA - metal electrode-electrolyte potential at the microelectrode tip

• EB - reference electrode - electrolyte potential
• EC - variable cell membrane potential

Figure (a) shows the position of the electrodes

Figure (b) is the electrical equivalent of metal microelectrodes

RA denotes the resistance of the connecting wire which is negligible.

Rs denote the resistance of the shaft of the microelectrode which is also negligible.

RFA, RWA, and CWA constitute the impedance of the microelectrode tip - intracellular fluid
interface; RIN is the resistance of the intracellular fluid and RB is the resistance of the wire
connected to the reference electrode.

RFB, RWB and CWB constitute the impedance of the reference electrode -extracellular fluid
interface and REX is the resistance of the extracellular fluid

CD is the distributed capacitance between the insulated shaft of the microelectrode and
extracellular fluid.

• The capacitance between the tip of the microelectrode and intracellular fluid is
negligible because of the potential difference across it does not change.
• The impedance of the microelectrode tip is inversely proportional to the area of the tip
and frequency.
• Thus when the input impedance of the amplifier is not high enough, it behaves as a
high pass filter.

Micropipet

• The non-metallic micropipet consists of a glass micropipet whose tip's diameter is

about 1 micrometer and its filled with a electrolyte usually 3 M KCL which is
compatible with the cellular fluids.
• A thin flexible metal wire from chlorided silver, stainless steel or tungsten is inserted
into the stem of the micropipet.
• The friction between the wire and the stem of the micropipet and the fluid surface
tension hold the micropipet on the wire.
• The other end of the metal wire is mounted to a rigid support and the other free end of
it in resting on the cell.

EA - potential between the metal wire and electrolyte filled in the micro-pipet.

EB - potential between the reference electrode and the extracellular fluid.

EC - variable cell membrane potential ED - potential exiting at the tip due to different
electrolytes present in the pipet and the cell.

E=EA+EB+ED.
 Figure shows the position of the electrodes

Figure - Electrical equivalent of micropipette

RA denotes the resistance of the connecting wire.

RFA, RWA, CWA constitute the impedance of the electrode electrolyte interface in the stem
of the micropipet.

RIN and REX is the resistance of the electrolyte inside the cell and the electrolyte outside the
cell.
RFB, RWB and CWB constitute reference electrode –electrolyte interface impedance

RB is the resistance of the wire connected to the reference electrode.

CD is the distributed capacitance existing between the fluid in the pipet and the extracellular
fluid.

C’D is the equivalent of distributed capacitance.

When the micro-pipet is coupled with the amplifier terminals A and B, then the membrane
potential Ec is coupled with it via a high series resistance ‘RT’ and a moderate shunt
capacitance CD along with electrode potentials.

2. Surface electrodes

Generally larger area surface electrodes are used to sense ECG potentials Smaller area
surface electrodes are used to sense EEG and EMG potentials.

Types of Surface electrodes

• Metal Plate electrode

• Suction cup electrode
• Adhesive tape electrode
• Multipoint electrode
• Floating electrode

Metal Plate electrode

Rectangular (3.5 cm x 5 cm) and circular (4.75 cm diameter) plates from German silver
nickel silver or nickel plated steel are used as surface electrodes in the case of ECG
measurement. When these electrodes are applied on the skin with electrode paste, typical DC
resistance values are in the range from 2 to 10 kilo ohms, the high frequency impedance
amounts to a few hundreds ohms.

Suction cup electrode

It is more practical and is well suited for attachment to flat surfaces of the body and to
regions where the underlying tissue is soft. Although physically large, this electrode has a
small area because only the rim is in contact with the skin.
Adhesive tape electrode

The pressure of the surface electrode against the skin may squeeze the electros paste out. To
avoid this problem, adhesive tape electrode is used. It consists of a light weigh metallic
screen backed by a pad for electrode paste as shown in figure. The adhesive backing holds
the electrode in place and retards the evaporation of the electrolyte present in the electrode
paste.

Multipoint electrode

The multipoint electrode is a very practical electrode for ECG measurements and it contains
nearly 1000 fine active contact points. By this a low resistance contact is established with the
subject. If the subject has hairs on the regions of interest, then one can use the multipoint
electrode without removing the hair. We can use it under any environmental conditions.
Floating electrode

In the floating electrode, the metal does not contact the subject directly. That is the contact is
made via an electrolytic bridge. By means of this electrode, movement artifact is eliminated.
This is also called as liquid junction electrode.

During the application of surface electrodes, we are getting signals from a relatively large
section of tissue. The activity we see is the total product of millions of nerve or muscle cells
working as a team. If it becomes necessary to evaluate the activity of a small section of tissue
or of cells themselves, then the surface electrodes are not useful. During long-term
monitoring or exercise testing the surface electrode is an important part of the system. The
pH of the electrode paste should be maintained at 7.0 during measurement and buffered

3. Needle electrodes

• Used to record nerve action potential.

• Used to measure EEG and EMG Signals.
• A short length of the fine insulated metal wire is bent at its one end and the bent
portion is inserted through the lumen of the needle and is advanced into the muscle.
• When we insert two insulated wires into the lumen of the needle, then the two wires
constitute bipolar electrode such that one wire act as active electrode and another act
as reference electrode.
• The main advantage of needle electrode is that they are less susceptible to movement
errors than surface electrode

Types of needle electrode

• Monopolar or Insulated Needle Electrode

• Bipolar Needle Electrode
• Concentric Or Coaxial Needle electrode

Monopolar or Insulated Needle Electrode

• Made of stainless steel, the monopolar needle electrode has a very finely sharpened
point and is covered with Teflon or other insulating material over its entire length,
except for a 0.5 mm exposure at the tip.
 • The needle serves as the active electrode, and a surface electrode placed on the skin
close to it serves as a reference.

• The main advantage of monopolar needle electrodes is small diameter and Teflon
covering allows them to slide in and out of the muscle easily.
• The major disadvantages of this needle are moving the needle causes less discomfort.
With repeated use, the size of the bare tip changes

Bipolar Needle Electrode

Bipolar needle electrodes contain two insulated wires within a metal cannula. Two wires are
bared at the tip and provide contacts to the patient. Bipolar electrodes are electrically
symmetrical and have no polarity sense.

Concentric Or Coaxial Needle electrode

• The concentric needle consists of a cannula with an insulated wire.

• The active electrode is the small tip of the center wire, and the reference electrode is
the outside cannula.
• Concentric needles may have two central wires (bipolar), in which case the active and
reference electrodes are at the tip and the outside cannula acts as the ground.

Advantage:

• Minimizes Background Noise

• No (Reference) Surface Electrode Is Needed.
Disadvantage:

• Larger diameter can cause more pain.

• Moving the electrode around is uncomfortable.
• Bending becomes a problem when the needle is dulled by repeated use.

Figure – needle electrode equivalent circuit

Applications of Needle Electrodes

• Needle electrodes are mostly used in the measurement of EEG and EMG signals.

10. RECORDING PROBLEMS

• Inaccessibility of variable to measurement: It is greatest difficulty in attempting

from a living system is the problem in gaining to the variable being measured. For
example neuro chemical activity of brain, it is impossible to place transducer so we
need to do the indirect measurement. By using indirect measurement, however one
must be aware of the limitations.
• Variability of data: Majority of physiological variables are nondeterministic, means
varies with respect to time.so these must be represented by some statistical or
probability distribution
• Lack of knowledge of interrelationship: physiological measurements with large
tolerance are often accepted by the physician because of lack of this knowledge and
the resultant in ability to control variations. Better understanding of physiological
relationship would also permit more effective use of indirect measurements as
substitutes for inaccessible measure.
• Transducer consists of two main parts, that is,
• Sensor or Sensing Element: This part is responsible for generating measurable
response with respect to the change in physical quantity to be measured.
• Transduction Element: Sensor output is carried on to the transduction element
which converts the non-electrical signal to electrical signal in proportion to the input
• Artifacts: it is component or variable is observed while doing experiment, which is
not naturally present. Thus random noise generated within the measuring instrument,
 electrical interference (50/60 Hz), cross talk and all other unwanted variations in a
signal are considered artifacts.
• Energy limitations: Many physiological measurement techniques that a certain
amount of energy be applied to the living system in order to obtain a measurement.
For example, resistance measurements require the flow of electric current through the
tissue or blood being measured. Some transducers generate small amount of heat due
to the current flow.
• Safety considerations: Methods employed in measuring variables in a living human
subject must in no way endanger the life or normal functioning of the subject. Recent
emphasis on hospital safety requires that extra caution must be taken in the design of
any measurement system to protect the patient.

11. MOTION ARTIFACTS

• When a polarizable electrode is in contact with an electrolyte, a double layer of

charge forms at the interface.
• If the electrode is moved with respect to the electrolyte, this movement mechanically
disturbs the distribution of charge at the interface and results in a momentary change
of the half-cell potential until equilibrium can be reestablished.
• If a pair of electrodes is in an electrolyte and one move while the other remains
stationary, a potential difference appears between the two electrodes during this
movement.
• This potential is known as motion artifact and can be serious cause of interference in
the measurement of biopotentials.
• Because motion artifact results primarily from mechanical disturbances of the
distribution of charge at the electrode-electrolyte interface, it is reasonable to expect
that motion artifact is minimal for nonpolarizable electrode.
• Observation of the motion-artifact signals reveals that a major component of this
noise is at low frequencies show that different biopotential signals occupy different
portions of the frequency spectrum.
• Low-frequency artifact does not affect signals such as the EMG or axon action
potential (AAP) nearly so much as it does the ECG, EEG, and electrooculogram
(EOG).
• In the former case, filtering can be effectively used to minimize the contribution of
motion artifact on the overall signal.
• But in the latter case, such filtering also distorts the signal. It is important in these
applications to use a non-polarizable electrode to minimize motion artifact stemming
from the electrode-electrolyte interface.
• This interface is not the only source of motion artifact encountered when biopotential
electrodes are applied to the skin.
• The equivalent circuit in electrode-skin interface, in addition to the half-cell potential
E, the electrolyte gel-skin potential Ehc can also cause motion artifact, if it varies with
movement of the electrode.
 • Variations in this potential represent a major source of motion artifact in Ag/AgCl
skin electrodes.
• Motion artifact can be significantly reduced when the stratum corneum is removed by
mechanical abrasion with a fine abrasive paper. This method also helps to reduce the
epidermal component of the skin impedance. They also point out that removal of the
body's outer protective barrier makes that region of skin more susceptible to irritation
from the electrolyte gel. Therefore, the choice of a gel material is important.

12. MEASUREMENTS WITH TWO ELECTRODES

• Measurement of the bioelectric potentials requires two electrodes.

• The voltage measured is really the difference between the instantaneous potential of
the two electrodes.

If two electrodes are of same type

• The difference is usually small and depends essentially on the actual difference of
ionic potential between the two points of the body from which measurements are
being taken.

If two electrodes are of different type

• They produce a significant dc voltage that can cause current to flow through both
electrodes as well as through the input circuit of the amplifier to which they are
connected.
• The DC voltage due to the difference in electrode potential is called offset voltage of
electrode the two Electrodes of same material may also produce small electrode offset
voltage.
• Chemical activity takes place within an electrode can cause voltage fluctuations to
appear without any physiological input. Such Variations may appear as noise on
bioelectric signal.
• It may reduce by proper choice of materials or by coating the electrodes to improve
stability, the best material for this is Silver-silver chloride.

Equivalent circuit for a pair of electrode

• The electrical equivalent circuit suggests that the voltage presented to the measuring
instrument from the electrode consists of two main components. One is the contact
potential and the other is the biological signal of interest.
• The contact potential depends upon several factors and may produce an interference
signal which exceeds several times the useful signal. The contact potential is found to
be a function of the type of skin, skin preparation and composition of the electrolyte.
• When bioelectric events are recorded, interference signals are produced by the
potential differences of metal-electrolyte and the electrolyte-skin interface.
• Normally, these potential differences are connected in opposition during the
recording procedure, and in the case of a truly reversible and uniform electrode pair,
their difference would be nil.
• However, in practice, and represent the half-cell potentials (E1 and E2) of the
electrodes.
• Z1 and Z2 are the skin contact impedances of these electrodes and R is the tissue
resistance or resistance of the bioelectric generator.

Figure - Equivalent circuit for a pair of electrode

• This circuit shows that the impedance of the electrodes would be high in the low
frequency region and it would decrease with increasing frequency.
• It is further clear that in the measurement of a bioelectric signal, it is essential to
minimize potential drops across the electrode impedance.
• This is achieved by making the skin-contact impedance as low as possible and making
the input impedance of the measuring device as high as possible.
 UNIT II BIOPOTENTIAL MEASUREMENTS

Bio signals characteristics - frequency and amplitude ranges. ECG - Einthoven's triangle,
standard 12 lead system, Principles of vector cardiography, EEG - 10-20 electrode system,
unipolar, bipolar and average mode, EMG - unipolar and bipolar mode and Recording of
ERG, EOG and EGG.

1. BIO SIGNALS CHARACTERISTICS - FREQUENCY AND AMPLITUDE

RANGES

Bioelectric Signals Frequency Voltage Range Electrode Used Origin

Range (Hz) (Micro
Voltage)
Electrocardiogram 0.05 to 100 10 to 5000 Surface electrodes are Heart muscles
(ECG) Vector covers fetal used with jelly, paste or
cardiogram range cream. Needle electrodes
are less noisy

Electroencephalogra 0.1 to100 2 to 200 Surface and Needle Neuronal

m (EEG) electrodes activity of the
brain

Electromyograph 5 to 2000 20 to 5000 Surface or Needle

(EMG) electrodes Skin muscles

Cerebral potentials Pulse duration 10 to 100000 Deep needle electrodes Cerebrum of the
0.6 ms to 0.1 s brain

Electrogastrogram 0.05 - 0.2 10- 350 Peristaltic

(EGG) Surface electrodes movements of
the
gestrointestinal
tract

Electroretinogram 0.1 to 100 10 - 350 Corneal electrodes Retina of the

(ERG) eye

Electrooculogram d.c to 100 10 to 3500 Miniature surface Corneal - retinal

(EOG) electrodes potential
variations
2. ECG - EINTHOVEN'S TRIANGLE

The closed path RA to LA to LL and back to RA is called the Einthoven triangle. According
to Einthoven, in the frontal plane of the body the cardiac electric field vector is a two
dimensional one. The ECG measured from any one of the three limb leads is a time variant
single dimensional component of that vector. Along the sides of this triangle the three
projections of ECG vector are measured. Further the vector sum of the projections on all the
three sides is equal to zero. Thus following Kirchoff's law, the R wave amplitude of lead II is
equal to the sum of the R wave amplitudes of leads I and III. For example the R wave
nominal voltage from different leads is given below.

The voltages given in brackets indicate the range of the measured voltage. Thus

3. ECG - STANDARD 12 LEAD SYSTEMS

• Electrocardiography is the method of recording an Electrocardiogram (ECG) with the

help of the machine Electrocardiograph.
• Electrocardiogram (ECG) is a graphic recording of the electrical changes that occur
within the heart during the cardiac cycle.
 Figure – Waveform of ECG

Uses of ECG

Electrocardiogram is useful in determining and diagnosing the following:

• Heart rate
• Heart rhythm
• Abnormal electrical conduction
• Poor blood flow to heart muscle (ischemia)
• MI
• Coronary artery disease
• Hypertrophy of heart chambers

Characteristics of ECG Waveform:

Waveforms Origin Amplitude Duration

mV Sec
P Wave Atrial Depolarisation or 0.25 0.12 to 0.22
Contraction (P-R Interval)
R-Wave Repolarisation of the atria or 1.60
(QRS Depolarisation of the ventricles 0.07 to 0.1
Complex)
T-Wave Ventricular Repolarisation 0.1 to 0.5
(Relaxation of Myocardium) 0.05 to 0.15
(S-T Interval)
S-T Interval Ventricular Contraction
- -
U-Wave Slow Repolarisation of <0.1
Intraventricular (Purkinje Fibre) 0.2
System (T-U Interval)
ECG LEADS

ECG Leads

Bipolar Limb Leads/standard Unipolar Leads

limb leads

Augmented Chest Leads

1. Lead I
Unipolar Limb
2. Lead II
Leads
3. Lead III

1. aVL 1. V1
2. aVL 2. V2
3. aVF 3. V3
4. V4
5. V5
6. V6

• ECG is recorded by placing series of electrodes on the surface of the body. These
electrodes are called ECG leads and are connected to the ECG machine.
• Electrodes are fixed on the limbs. Usually, right arm, left arm and left leg are chosen.
Heart is said to be in the center of an imaginary equilateral triangle drawn by
connecting the roots of these three limbs. This triangle is called Einthoven triangle.
• ECG is recorded in 12 Leads.
• Classified into two types:

1. Bipolar Leads

2. Unipolar Leads

BIPOLAR LIMB LEADS

In standard leads, the potentials are tapped from four locations of our body. are (i) right arm,
(ii) left arm, (iii) right leg and (iv) left leg. Usually the right leg electrode is acting as ground
reference electrode.

Bipolar limb leads are otherwise known as standard limb leads. Two limbs are connected to
obtain these leads and both the electrodes are active recording electrodes, i.e. one electrode is
positive and the other one is negative.
Standard limb leads are of three types:

a. Limb lead I

b. Limb lead II

c. Limb lead III

Lead I

Lead I is obtained by connecting right arm and left arm. Right arm is connected to the
negative terminal of the instrument and the left arm is connected to the positive terminal.
Lead I Position gives voltage V1, the voltage drop from the left arm LA to the right arm RA.
Lead II

Lead II is obtained by connecting right arm and left leg. Right arm is connected to the
negative terminal of the instrument and the left leg is connected to the positive terminal. Lead
II Position gives voltage VII, the voltage drop from the left leg LL to the right arm RA.

Lead III

Lead III is obtained by connecting left arm and left leg. Left arm is connected to the negative
terminal of the instrument and the left leg is connected to the positive terminal. Lead III
Position gives voltage VIII, the voltage drop from the left leg LL to the left arm LA.

The voltages given in brackets indicate the range of the measured voltage. Thus

UNIPOLAR LEADS

Here, one electrode is active electrode and the other one is an indifferent electrode. Active
electrode is positive and the indifferent electrode is serving as a composite negative electrode.

Unipolar leads are of two types:

1. Unipolar limb leads

2. Unipolar chest leads.

Unipolar Limb Leads

• In the augmented unipolar limb leads system, which is introduced by Wilson, the
electrocardiogram is recorded between a single exploratory electrode and the central
terminal which has a potential corresponding to the center of the body.
• Thus two equal and large resistors are connected to a pair of limb electrodes and the
center of this resistive network acts as central terminal and the remaining limb
electrode acts as the exploratory electrode. By means of augmented ECG lead
connections, a small increase in the ECG voltage can be realized.
• The augmented lead connections are augmented voltage Right arm (aVR), augmented
voltage Left arm (aVL) and augmented voltage Foot (aVF).
 • Unipolar limb leads are also called augmented limb leads or augmented voltage leads.
Active electrode is connected to one of the limbs. Indifferent electrode is obtained by
connecting the other two limbs through a resistance.
Unipolar limb leads are of three types:

i. aVR lead

ii. aVL lead

iii. aVF lead

i. aVR lead

Active electrode is from right arm. Indifferent electrode is obtained by connecting left arm
and left leg.

ii. aVL lead

Active electrode is from left arm. Indifferent electrode is obtained by connecting right arm
and left leg.

iii. aVF lead

Active electrode is from left leg (foot). Indifferent electrode is obtained by connecting the
two upper limbs.

Unipolar Chest Leads

• In the case of unipolar chest leads, the exploratory electrode is obtained from one of
the chest electrodes. The chest electrodes are placed on the six different points on the
hest closed to the heart.
• By connecting three equal large resistances to the left arm, right arm and left leg a
reference electrode or central terminal obtained.

• This lead system is known as Wilson system. Thus the electrocardiograms are
recorded from these 12 lead selections such that 3 standard bipolar leads, 3 augmented
unipolar leads and 6 chest leads.
 • Chest leads are also called 'V' leads or precardial chest leads.
• Indifferent electrode is obtained by connecting the three limbs, viz. left arm, left leg
and right arm, through resistances.
• Active electrode is placed on six points over the chest.
• This electrode is known as the chest electrode and the six points over the chest are
called V1, V2, V3, V4, V5, and V6.
• V indicates vector, which shows the direction of current flow.

ECG recording setup:

Patient cable

The patient cable connects the different leads from the limbs and chest to the defibrillator
protection circuit.

Defibrillator Protection Circuit

The one end of the electrode leads are connected along RA, LA, chest and LL of the patient.
The other end of electrode passes through defibrillator protection circuit. The protection
circuit has buffer amplifier and over-load voltage protection circuit.

Lead Selection Logic

This block helps to select the type of electrode lead system. We can choose either bipolar or
augmented electrode system.

Calibration Circuit

Calibration is a process that helps to eliminate errors in the system. Here, any changes in the
lead selection circuit results in artifacts in the ECG output. Therefore, the calibration unit
helps the technician to correct the error in ECG output.

Bio-amplifier

The bio-amplifier consists of a preamplifier, power amplifier, instrumentation amplifier and

differential amplifier with high gain and high CMRR. The sensitivity or the gain of the
amplifier can be varied. Followed by the preamplifier, there is a power amplifier which is
used to drive the recorder. Pen motors in the recorder requires sufficient electrical power to
activate the recording or display. Therefore power amplifiers are required with high power
gain.

Auxiliary amplifier

Since the electrode impedances are not equal, a differential amplifier does completely reject
the common mode signals. The common mode signals can be reduced a minimum level by
means of adding an auxiliary amplifier between the driven right leg le and the ECG unit. By
this way, the right leg is not connected to ground but it is connect to the output of the
auxiliary amplifier.
 Figure – block diagram of ECG recording setup

Isolated Power Supply

The isolated power supply is used to give power to the bio-amplifier and by means of that,
the electrical safety for the patient is increased.

Output Unit

The output unit is a cathode ray oscilloscope or a paper chart. In the case of paper chart
recorder, the power amplifier or pen amplifier supplies the required power to drive pen motor
that records the ECG trace on the wax coated hear sensitive paper. The paper speed is about
25 mm/s (U.S. paper speed) or 50 mm/s (European paper speed). The faster speed of 50 mm/s
is provided to allow better resolution of the ORS complex at very high heart rates.

Power Switch

The power switch of the recorder has three positions. In the ON position the power to the
amplifier is turned on; but the paper drive is not running. In order to start the paper drive the
switch must be placed in the RUN position. In OFF position, the ECG unit is in switched off
condition. The power switch of the recorder has three positions. In the ON position the power
to the amplifier is turned on; but the paper drive is not running. In order to start the paper
drive the switch must be placed in the RUN position. In OFF position, the ECG unit is in
switched off condition.

4. PRINCIPLES OF VECTOR CARDIOGRAPHY

• Vectorcardiography (VCG) is a diagnostic tool used in cardiology to analyze the

electrical activity of the heart and create a graphical representation of the heart's
electrical vectors. These vectors represent the direction and magnitude of the
electrical signals generated by the heart during each cardiac cycle.
• The electrical activity of the heart is primarily due to the depolarization and
repolarization of cardiac muscle cells. These electrical changes can be measured by
placing multiple electrodes on the body's surface, typically in specific configurations.
The most common setup is the Frank lead system, which uses three orthogonal leads
to record electrical signals from the heart.
• The three orthogonal leads in the Frank lead system are:
1. Lead X: Represents the horizontal electrical activity (left-right direction).
2. Lead Y: Represents the vertical electrical activity (up-down direction).
3. Lead Z: Represents the anterior-posterior electrical activity (front-back
direction).
• The electrical potentials from the three orthogonal leads (X, Y, and Z) are converted
into their corresponding vector components. For example, the X, Y, and Z leads may
represent the horizontal, vertical, and anterior-posterior directions, respectively.
• Using the converted lead data, the magnitude and direction of the cardiac electrical
vectors are calculated. This step involves vector mathematics to determine the
resultant vector of the heart's electrical activity during each phase of the cardiac cycle.
• The calculated vectors are used to construct the vectorcardiogram. A
vectorcardiogram is typically a graphical representation of the heart's electrical
activity in 3D space. It may be represented as loops or loops within loops, known as
"vector loops" or "cardiac loops.

5. EEG - UNIPOLAR, BIPOLAR AND AVERAGE MODE

• An electroencephalogram (EEG) is a test that measures electrical activity in the brain

using small, metal discs (electrodes) attached to the scalp.

• Brain cells communicate via electrical impulses and are active all the time, even
during asleep.

• An EEG can find changes in brain activity that might be useful in diagnosing brain
disorders, especially epilepsy or another seizure disorder.

• Uses of EEG

An EEG might also be helpful for diagnosing or treating:

• Brain tumors
• Brain damage from head injury

• Brain dysfunction that can have a variety of causes (encephalopathy)

• Sleep disorders

• Inflammation of the brain

• Stroke

Figure – Brain Lobe

BRAIN LOBE

Frontal Lobe

• The frontal lobe is generally where higher executive functions including emotional
regulation, planning, reasoning and problem solving occur.

Parietal Lobe

• Areas in the parietal lobe are responsible for integrating sensory information,
including touch, temperature, pressure and pain.

Temporal Lobe

• The temporal lobe contains the primary auditory cortex, which receives auditory
information from the ears.

Occipital Lobe

• The occipital lobe is the major visual processing centre in the brain

Brain waves

• Brain waves are patterns of electrical activity occurring in the brain.

Types of brain waves

 • Alpha waves
• Beta waves
• Delta brain
• Theta waves
• Gamma brain waves
1. Alpha waves
• Freq: 8-13HZ
• Occurance: they found in normal person when they are awake in quiet and resting
state.
2. Beta waves

• Freq : 13-30 HZ
• Occurance: it occurs a mental activity like tension –freq increases upto 50HZ.
3. Theta wave:

• Freq: 0.5-4HZ
• Occurance: recorded from parietal & temporal regious of scalp of children. occurs
during emotional stress in some adults, particularly during disappointments &
frustrations.
4. Delta wave :

• Frequency:0.5-4 Hz
• Occurance: The occurs in every 2 or 3 seconds. occurs in deep sleep ,in premature
babies and in very serious brain diseases

Block Diagram of EEG Recording Setup

• Patient cable consists of 21 electrodes and is connected to the 8 channel selector.

• The electrodes are attached to the channel selector in groups of 8 Called Montage of
electrodes.
• The right ear electrodes acts reference electrode for a right brain electrodes and left
ear act as reference electrode for left brain electrode.
• Output from 8 channel connecter goes to differential Amplifier. The output voltage
from the amplifier may either be applied directly to time eight channel display
through filter bank or it may be stored on data on a tape recorder or computer memory
for further processing.

Figure –Block diagram of EEG recording setup

• The system helps to record the potentials generated from Sensory parts of the brain.
To achieve this, output unit is connected with audio, visual and touch stimulus.
• It can also measure the time delay between stimulus and response from brain. In
addition, we have a filter bank- Consist of LP, HP and BP filters, they help to
removenoise from the brain waves.
• For the output recording, we can see either pen recorder of CRO.
• Three modes rarely unipolar, bipolar and wilson mode or average mode recording are
used to measure EEG.
• In bipolar technique, the difference in potential between two adjacent electrodes is
measured.
• In the monopolar technique, the potential of each electrode is measured with respect
to a reference electrode attached to ear lobe.
• In the Wilson technique (or) average mode recording techniques the potential is
measured between one of the electrodes (exploring electrode) and the central terminal
which is formed by connecting all electrodes through high, equal resistors to a
common point.
Applications of EEG
EEG helps physicians to diagnose
• The level of consciousness
• Sleep disorders
• Brain death
• Brain tumors
• Epilepsy
• Multiple sclerosis

6. EEG – 10-20 LEAD SYSTEM

• The 10/20 system or International 10/20 system is an internationally recognized

method to describe the location of scalp electrodes.
• The system is based on the relationship between the location of an electrode and the
underlying area of cerebral cortex.
• The numbers ‘10’ and ‘20’ refer to the fact that the distances between adjacent
electrodes are either 10% or 20% of the total front- back or right-left distance of the
skull.
• Each site has a letter to identify the lobe and a number to identify the hemisphere
location.

Electrode Lobe

F Frontal

T Temporal

C Central *

P Parietal

O Occipital

Placement of electrodes

In EEG, electrodes are placed in standard positions on the skull in an arrangement called 10-
20 system, a placement scheme devised by the International Federation of Societies of EEG.
The electrodes in this arrangement are placed as follows:

• Draw a line on the skull from the nasion, the root of the nose, to the inion, ossification
center (bump) on the occipital lobe.
• Draw a similar line from the left preauricular (ear) point to the right preauricular
point.
• Mark the intersection of these two lines as Cz which is the mid point of the distance
between the nasion and inion (or) the distance between the auricular points.
• Mark points at 10, 20, 20, 20 and 10% of the total nasion-inion distance. These points
are Fpz, Fz, Cz, Pz and Oz.

• Mark points at 10, 20, 20, 20, 20 and 10% of the total distance between the
preauricular points. These points are T3, C3, Cz, C4 and T4. In these odd numbered
points T3 and C3 are on the left and even numbered points C4 and T4 are on the right.
• Measure the distance between Fpz and Oz along the great circle passing through T3
and mark points at 10, 20, 20, 20, 20 and 10% of this distance. These are the positions
of Fp1, F7, T3, T5 and O1.
• Repeat this procedure on the right side and mark the positions of Fp2, F8, T4, T6 and
O2.
• Measure the distance between Fp1 and O1 along the great circle passing through C3
and mark points at 25% intervals. These points give the positions of F3, C3 and P3.
The ground reference electrode is a metal clip on the earlobe,
• Repeat this procedure on the right side and mark the positions of F4, C4 and P4.
• Check that F7, F3, Fz, F4 and F8 are equidistant along the transverse circle passing
through F7, Fz and F8 and check that T5, P3, Pz, P4 and T6 are equidistant along the
transverse circle passingT5, Pz and T6. In the figure, the positions of the scalp
electrodes are indicated. Further there are nasopharyngeal electrodes Pg1 and Pg2 and
ear electrodes A1 and A2.

7. EMG - UNIPOLAR AND BIPOLAR MODES

 • Electromyography is the science of recording and interpreting the electrical activity of
muscle's action potentials. Meanwhile the recording of the peripheral nerve's action
potentials is called electro-neurography.
• Electromyograph is an instrument used for recording the electrical activity of the
muscles to determine whether the muscle is contracting or not.
• The electrical activity of the underlying muscle can be measured by placing surface
electrodes on the skin. To record the action potentials of individual motor units, the
needle electrode is inserted into the muscle.
• Thus EMG indicates the amount of activity of a given muscle or a group of muscles
and not an individual nerve fiber.
• The action potentials occurs both positive and negative polarities at a given pair of
electrodes; so they may add or cancel each other.
• The contraction of a muscle produces action potentials. When there is stimulation to a
nerve fiber, all the muscle fibers contract simultaneously developing action potentials.
In a relaxed muscle, there is no action potential.
• EMG measurements are also important for the myoelectric control of prosthetic
devices (artificial limbs).

Types of electrodes are used for recording EMG.

1. Surface electrodes

• The electrical activity of the underlying muscle can be measured by placing

surface electrodes on the skin.

2. Needle electrodes

• To record the action potentials of individual motor units, the needle electrode is
inserted into the muscle.

The block diagram shows a typical set-up for EMG recordings.

• EMG is usually recorded by using surface electrodes or more often by using needle
electrodes, which are inserted directly into the muscle.
• The surface electrodes may be disposable, adhesive types or the ones which can be
used repeatedly.
 • A ground electrode is necessary for providing a common reference for measurement.
These electrodes pick up the potentials produced by the contracting muscle fibres.
• The signal can then be amplified and displayed on the screen of a cathode ray tube.
• It is also applied to an audio amplifier connected to a loudspeaker. A trained EMG
interpreter can diagnose various muscular disorders by listening to the sounds
produced when the muscle potentials are fed to the loudspeaker.
• The oscilloscope displays EMG waveforms.

Figure - Block diagram of a typical set-up for EMG recording

• The tape recorder is included in the system to facilitate playback and study of the
EMG sound waveforms at a later convenient time.
• The waveform can also be photographed from the CRT screen by using a
synchronized camera.

• The amplitude of the EMG signals depends upon various factors, e.g. the type and
placement of electrodes used and the degree of muscular exertions.
• The needle electrode in contact with a single muscle fibre will pick up spike type
voltages whereas a surface electrode picks up many overlapping spikes and therefore
produces an average voltage effect.
• A typical EMG signal ranges from 0.1 to 0.5 mV. They may contain frequency
components extending up to 10 kHz. Such high frequency signals cannot be recorded
on the conventional pen recorders and therefore, they are usually displayed on the
CRT screen.

Types of EMG Configuration

• Mono-polar configuration
• Bi-polar configuration
• Multi-polar configuration
Mono-polar configuration / unipolar configuration

The mono-polar configuration is implemented using only a single electrode on the skin with
respect to a reference. This method is used because of its simplicity.

Bi-polar configuration

• Bipolar configuration is used to acquire EMG signal using two EMG detecting
surfaces with the help of a reference electrode.
• The signals from the two EMG surfaces are connected to a differential amplifier. The
two detecting surfaces are placed only 1-2 cm from each other.

Multi-polar configuration

• This configuration uses more than two detecting surfaces to acquire the EMG signal
with the help of a reference electrode and further reduces crosstalk and noise
concerns.
• The signals from three or more EMG detecting surfaces, placed 1-2 cm from each
other, are passed through more than two stages of differential amplification.
• For example if three detecting surfaces are used then double differential technique is
employed.

Application of EMG

• To study neuromuscular functions.

• To know the neuromuscular condition.
• To study reflex responses.
• Helps to diagnose the muscular diseases
 8. RECORDING OF EOG

Electrooculography

• Electrooculography (EOG) is a non-invasive technique used to measure the electrical

activity generated by the muscles surrounding the eyes.
• The electrical signals produced by eye movements are recorded through electrodes
placed on the skin near the eyes.
• EOG is widely used in various applications, such as medical diagnostics, sleep
studies, eye movement studies, and brain-computer interface research.

Principle of EOG:

• EOG works based on the principle that the eyes have a positively charged cornea and
a negatively charged retina, creating a dipole between them.
• When the eyes move, the distribution of these charges changes and this generates
electrical potentials around the eyes.
• By measuring these changes in electrical potentials using electrodes, we can
determine the direction and magnitude of eye movements.

Techniques in EOG:

There are several EOG techniques used to record eye movement signals. The two
main techniques are:

Monopolar EOG:

• This technique involves placing one electrode around each eye, and a reference
electrode is placed at a distant location, typically on the forehead.
• The voltage difference between each eye electrode and the reference electrode is
measured, providing information about vertical and oblique eye movements.

Bipolar EOG:

• In this technique, two electrodes are placed around each eye, forming a bipolar
configuration.
• One electrode is placed close to the outer eye, and the other is placed near the outer
eye.
• The voltage difference between these two electrodes is measured as the eye moves,
providing information about horizontal eye movements.
 Figure – EOG Electrode Placement

Uses of EOG:

EOG has various applications in both medical and research settings:

• Sleep Studies: EOG is used to monitor eye movements during sleep studies to
identify different stages of sleep and rapid eye movement (REM) sleep.
• Eye Movement Studies: EOG is used to analyze eye movement patterns and study
ocular motor function, including saccades, smooth pursuit, and fixation.
• Brain-Computer Interface (BCI) Research: EOG is used as an input signal in BCI
systems, enabling individuals to control external devices using their eye movements.
• Ophthalmology: EOG is used to diagnose eye movement disorders.
• Human-Computer Interaction: EOG can be used as an alternative input method for
individuals with limited motor function, allowing them to interact with computers or
assistive devices using eye movements.

The block diagram of an Electrooculography (EOG) system

Electrodes: The electrodes are placed around the eyes to detect the electrical potentials
generated by eye movements. Usually, two or more electrodes are used, depending on the
specific EOG technique employed (bipolar or monopolar).

Signal Conditioning: The raw electrical signals obtained from the electrodes are weak and
noisy. Signal conditioning involves amplifying and filtering the signals to enhance their
quality and make them suitable for further processing.

Analog-to-Digital Converter (ADC): The conditioned analog signals are converted into
digital format using an ADC. This step is essential for digital signal processing and analysis.

Signal Processing: Digital signal processing techniques are applied to the EOG signals to
extract relevant features and information about eye movements. Various algorithms are used
to detect saccades, blinks, and other eye movement patterns.

Eye Movement Analysis: The processed signals are analyzed to determine the direction and
magnitude of eye movements, such as horizontal, vertical, and oblique movements.
 Signal conditioning ADC DSP

Application Specific Data Display and

Output and Control
Processing Visualization

Figure - Block diagram of an Electrooculography (EOG)

Data Display and Visualization: The results of the eye movement analysis are displayed
and visualized, typically in the form of graphs or eye movement traces, allowing researchers
or clinicians to interpret and understand the data.

Application Specific Processing: Depending on the specific application, additional

processing may be performed, such as sleep stage identification in sleep studies or eye
movement pattern analysis in ophthalmology.

Output and Control: The EOG system may provide output signals or control signals for
various applications, such as interfacing with a brain-computer interface (BCI) or controlling
external devices based on eye movements.

9. RECORDING OF ERG

An electroretinogram (ERG) is a diagnostic test used to measure the electrical activity of the
retina in response to light stimulation. It is a valuable tool for assessing the function of the
retina and identifying any abnormalities or disorders affecting the visual system. The normal
response pattern has waves called a wave and b wave.

A-wave: Both rods and cones contributes to it. This is produce due to the hyper-polarisation
of rod and cones B-wave: The b-wave is generated by ON and OFF bipolar cell (B) activity.

Figure – Waveform of ERR

Types of ERG

Full-Field ERG: This is the most common type of ERG, where a full-field stimulus (a
flashing light or pattern) is presented to the entire retina. Full-field ERG measures the overall
retinal response and is useful for diagnosing and monitoring general retinal health, as well as
detecting abnormalities such as retinal dystrophies and other retinal disorders.

Pattern ERG (PERG): In this type of ERG, a specific patterned stimulus, often a black-and-
white checkerboard pattern, is presented to the central visual field. PERG assesses the
function of retinal ganglion cells and their responses to patterned stimuli. It's particularly
helpful in diagnosing and monitoring diseases affecting the optic nerve, such as glaucoma.

Multifocal ERG (mfERG): In mfERG, multiple smaller stimuli are presented simultaneously
to different regions of the retina. This technique provides spatial information about the retinal
response and can help localize areas of dysfunction. It's useful for detecting early stages of
diseases like macular degeneration and diabetic retinopathy.

Electrodes Placement:

Figure – ERG Electrode placement

Electrodes are placed on or near the eye to record the electrical responses of the retina. The
most commonly used electrodes are:

a. Corneal Electrode: A contact lens-type electrode is placed on the cornea. This electrode is
in direct contact and picks up electrical signals from the retina.

b. Skin Electrodes: Small adhesive skin electrodes are attached around the eye on the
forehead. These electrodes measure the electrical potentials generated by the retina.

Block diagram of ERG:

Light Stimulator: During an ERG test, the patient is exposed to flashes of light or patterns
of light that are carefully controlled in intensity and duration.
Electrodes: Electrodes are placed on the surface of the eye or around the eye to detect the
electrical responses of the retina. The most commonly used electrodes are corneal electrodes
(contact lens-type electrodes) or skin electrodes placed on the skin around the eye.

Retinal Response: When the retina is exposed to light, the photoreceptor cells (rods and
cones) in the retina are stimulated and produce electrical responses. These responses are
transmitted through the optic nerve to the visual processing centres in the brain.

Amplification: The electrical responses from the electrodes are extremely small and require
amplification for accurate measurement.

Filtering: The amplified electrical response may contain noise or artifacts that need to be
filtered.

Figure - Block diagram of ERG

Analog-to-Digital Converter (ADC): The analog signals are then converted into digital
signals using an ADC.

Data Display: The recorded data is analyzed to assess the overall health and function of the
retina.

10. RECORDING OF EGG

The term "electrogastrogram" refers to a procedure used to record the electrical activity of the
stomach. The stomach has its own electrical rhythm, which controls the contractions of the
muscles and helps with the digestion process.

The EGG is performed using electrodes that are attached to the skin on the abdomen. These
electrodes detect the electrical signals generated by the stomach's muscles as they contract
and move. The recorded signals are then amplified, filtered, and displayed on a graph or chart
called an electrogastrogram.
EGG Electrode Placement

During an electrogastrogram, electrodes are placed on the surface of the skin over the
stomach region. These electrodes detect and record the electrical signals generated by the
stomach muscles. Typically, six electrodes are used for an EGG recording.

Figure - EGG Electrode Placement

Conditions of EGG

• A bradygastriais defined, how decreased rate of electrical activity in the stomach, as less
than 2 cycles per minute for at least 1 minute

• A tachygastria is defined, how increased rate of electrical activity in the stomach, as more
than 4 cycles per minute for at least 1 minute.

Block diagram of EGG:

EGG Electrodes: Surface electrodes are placed on the patient's abdominal area. These
electrodes pick up the electrical signals generated by the stomach muscles.

EGG Amplifier: The electrical signals detected by the electrodes are weak and need to be
amplified for accurate measurements. An amplifier is used to increase the strength of these
signals.
Analog-to-Digital Converter (ADC): The amplified analog signals are then converted into
digital signals using an ADC. This conversion is necessary to process the data using digital
signal processing techniques.

Figure - Block diagram of EGG

Data Analysis Unit: The recorded data can be analyzed using specialized software to extract
relevant information about gastric function.

Data Display System: The processed signals are displayed on a screen for real-time
visualization and monitoring. Additionally, the data can be recorded for further analysis.

Uses of Electrogastrogram (EGG)

Diagnosis of Gastrointestinal Disorders: Abnormalities in stomach muscle contractions and

rhythms can be detected through EGG readings.

Assessment of Gastric Motility: EGG helps medical professionals understand the motility
patterns of the stomach, which can aid in evaluating the movement and coordination of the
digestive system.

Research Tool: EGG is utilized in research studies to investigate the mechanisms of various
gastrointestinal disorders, helping researchers better understand the physiological and
neurological aspects of stomach function.

Guiding Surgical Interventions: In some cases, EGG might be used to assist surgeons in
planning procedures that involve the stomach or the digestive tract, ensuring a better
understanding of the patient's underlying condition.

Evaluation of Nausea and Vomiting: EGG can be used to study the relationship between
stomach activity and sensations of nausea and vomiting, providing insights into potential
treatments for these symptoms.

Pediatric Applications: EGG can be used to assess gastric motility in pediatric patients,
helping diagnose and manage gastrointestinal disorders in children.
UNIT III SIGNAL CONDITIONING CIRCUITS
Need for bio-amplifier - single ended bio-amplifier, differential bio-amplifier, Impedance
matching circuit, isolation amplifiers - transformer and optical isolation - isolated DC amplifier
and AC carrier amplifier., Power line interference, Right leg driven ECG amplifier, Band pass
filtering.
NEED FOR BIO-AMPLIFIER
Bio amplifiers
Generally, Bio signals are having low amplitude and low frequency. Amplifiers are needed to
boost the amplitude level of the bio signals. The output of this amplifier is displayed as EEG
or ECG waveform. These amplifiers are known as bio amplifiers or bio medical amplifiers.
Need of Bio Amplifier:
1. The biological amplifier should have a high input impedance value. The range
of value lies between 2 MΩ and 10 MΩ depending on the applications. Higher
impedance value reduces distortion of the signal.
2. When electrodes pick up biopotentials from the human body, the input circuit
should be protected. Every bio-amplifier should consist of isolation and
protection circuits, to prevent the patients from electrical shocks.
3. Since the output of a bioelectric signal is in millivolts or microvolt range,
the voltage gain value of the amplifier should be higher than 100dB.
4. Throughout the entire bandwidth range, a constant gain should be maintained.
5. A bio-amplifier should have a small output impedance.
6. A good bio-amplifier should be free from drift and noise.
7. Common Mode Rejection Ratio (CMRR) value of amplifier should be greater
than 80dB to reduce the interference from common mode signal.
8. The gain of the bio-amplifier should be calibrated for each measurement.

SINGLE ENDED BIO-AMPLIFIER

A single-ended amplifier is an amplifier circuit configuration where there is only one active
device (commonly a vacuum tube or transistor) in the signal path, and all other circuit elements
serve to provide power to this device. The output is taken directly from the active device, hence
the name ‘single-ended’. The single-ended amplifier is the simplest of all amplifier circuit
configurations.
Inverting amplifier:
An inverting amplifier (also known as an inverting operational amplifier or an inverting op-
amp) is a type of operational amplifier circuit which produces an output which is out of phase
with respect to its input by 180o.

The voltage gain of the inverting operational amplifier or inverting op amp is,

Non inverting amplifier:

A non-inverting amplifier is an OPAMP circuit configuration whose output is in phase with
the input signal at the non-inverting input. The input signal is applied at the non-inverting input
of the opamp. A non-inverting amplifier also acts as a voltage follower circuit.

The non-inverting amplifiers also have negative feedback which is used to control the gain of
the amplifier. Feedback contains a voltage divider circuit that provides a part of the output to
the input terminal. This OPAMP has a high input impedance and a low output impedance.

DIFFERENTIAL BIO-AMPLIFIER
A differential amplifier is a type of electronic amplifier that amplifies the difference between
two input voltages and rejects the average or common mode value of the two voltages.
It has two inputs and one output in which the output is ideally proportional to the difference
between the two voltages.

V0 is the output voltage

V1 and V2 are the input voltages
Ad is the gain of the amplifier (i.e. the differential amplifier gain)

when V1 = V2, the output voltage V0 is equal to zero, and hence the output voltage is
suppressed. But any difference between inputs V1 and V2 is amplified by the differential
amplifier gain Ad.

Differential amplifier is also known as a difference amplifier because the difference between
the input voltages is amplified.

Figure – Differential Amplifier

Hence its output voltage will be equal to the sum of the output voltages produced by the Op-
Amp circuit operating as an inverting amplifier and the Op-Amp circuit operating as a non-
inverting amplifier. Thus, one gets:

Now, if R1 = R2 and R3 = Rf, then:

Differential gain:
Differential-mode input voltage is the voltage difference between V1 and V2.
Differential mode can readily act as a subtractor amplifier as it results in an output voltage
given by:

Where V1 and V2 represent the voltages applied at its inverting and non-inverting input
terminals and Ad refers to its differential gain.

Common mode gain(Ac):

Common-mode input voltage is the average value of V1 and V2.
Vout = Ac * Vc

Vc =

The output of an ideal differential amplifier is given by

Modes of operation of Differential Amplifier (DA)

There are two modes of operations of DA
1. Differential mode
2. Common mode
Differential mode
Two inputs are of equal in magnitude and different phase are used.
Common mode
Two inputs are of equal in magnitude and same phase are used.
Common mode rejection ratio
The common-mode rejection ratio (CMRR), usually defined as the ratio between differential-
mode gain and common-mode gain, indicates the ability of the amplifier to accurately cancel
voltages that are common to both inputs. The common-mode rejection ratio is defined as
IMPEDANCE MATCHING CIRCUIT
Impedance matching is defined as the process of designing the input impedance and
output impedance of an electrical load to minimize the signal reflection or maximize the power
transfer of the load. This source impedance is equivalent to resistance in series with reactance.
According to the maximum power transfer theorem, when the load resistance is equal
to the source resistance and load reactance is equal to negative of the source reactance, the
maximum power is transferred from source and load. It means that the maximum power can
be transfer if the load impedance is equal to the complex conjugate of the source impedance.

Consider that, the Resistor (R) and Inductor (L) are in series. And this combination is
in parallel with the Capacitor (C). Hence, the Impedance is,

Block diagram of Impedance matching

Source: This represents the signal source. Source" represents the device that generates the
signal.
Impedance Matching Circuit. It typically consists of passive components like resistors,
capacitors, and inductors arranged in a specific configuration. Its purpose is to adjust the
impedance of the source or load to match the other side.
Load: "Load" is the device that receives the signal.

Condition of impedance matching

ZS is donated as source impedance. ZL is donated as load impedance.
1. ZS = ZL
If the source impedance is equal to the load impedance, the maximum power will transfer from
source to load.
2. ZS ≠ZL
If the source impedance is not equal to the load impedance, the maximum power will not
transfer from source to load. The signal reflection will occur.
Types of impedance matching
1. Transform matching:
RF transformers can be used to produce very wideband impedance matching. The main
limitations are the restricted range of available impedances and the frequency limitations on
transformers.
2. LC matching:
LC matching where inductors and capacitors are used to affect the impedance transformation.
LC matching results in a relatively narrow bandwidth match. LC matching is very practical at
frequencies from 30mHz to 300Mhz. LC matching permits easy tuning of the match to allow
for device variations.

ISOLATION AMPLIFIERS - TRANSFORMER AND OPTICAL ISOLATION

Isolation amplifiers
Isolation amplifiers are known as Pre-amplifier isolation circuits. An isolation amplifier
increases the input impedance of a patient monitoring system. It also helps to isolate the patient
from the device. Using the isolation amplifier prevents accidental internal cardiac shock.

The electrical signals are obtained with electrodes. The signals received goes to the amplifier
block, where signals amplification occurs. After amplification, the signal enters the modulation
block. When either it goes to the isolation barrier, optical cable or transformer can be used. If
in case of optical cable, modulator output travels to LED. The LED converts electrical signals
into light energy. If the transformer acts an isolation barrier, modulator output connects the
primary winding of the transformer. Energy from primary transfers to the secondary winding
based on the mutual induction principle. At the next stage, secondary output enters the
demodulation block. Finally, the amplified demodulated signal is obtained.
Types of isolation amplifiers
1. Transformer isolation amplifiers
2. Optical isolation amplifiers
3. Capacitive isolation amplifiers
Transformer isolation amplifier
A transformer -isolated amplifier relies on transformer coupling of a high-frequency carrier
signal between input and output. Some models also include a transformer-isolated power
supply, that may also be used to power external signal processing devices on the isolated side
of the system.
An isolation transformer is a transformer used to transfer electrical power from a source of
alternating current (AC) power to device while isolating the powered device from the power
source.
In theory, the definition of 'isolation transformer' applies to any transformer where there is no
direct connection between the primary and the secondary windings. The windings are
connected only by the magnetic flux in the core.

The electrical signals are obtained with electrodes. The signals received goes to the amplifier
block, where signals amplification occurs. After amplification, the signal enters the modulation
block. When either it goes to the isolation barrier transformer can be used. If the transformer
acts an isolation barrier, modulator output connects the primary winding of the transformer.
Energy from primary transfers to the secondary winding based on the mutual induction
principle. At the next stage, secondary output enters the demodulation block. Finally, the
amplified demodulated signal is obtained.
Optical isolation amplifier
An optically isolated amplifier modulates current through an LED optocoupler. The linearity
is improved by using a second optocoupler within a feedback loop. Some devices provide up
to 60 kHz bandwidth.
In isolation amplifier the coupling method between input and output is optical. The most often
isolation amplifier contains an input amplifier of LED and a silicon photodiode an output
amplifier. The following figure shows such configuration of isolation amplifier.

Input and outputs are electrically isolated from each other. They are optically coupled. LED
emits light and it is detected by a photodiode at output.
According to input signal, light emitted by LED changes i.e. intensity of light is variable. This
is detected by a photodiode at output and generates a photocurrent at output. Therefore, light
emitted is proportional to photocurrent generated. The signal from output is further amplified
by amplifier. The signal from input is optically transferred to output side providing electrical
isolation.

ISOLATED DC AMPLIFIER
An isolated DC amplifier is a device used to amplify a direct current (DC) signal while
providing electrical isolation between the input and output. This isolation helps prevent noise,
ground loop issues, and other interference from affecting the signal. The primary goal is to
maintain the integrity of the DC signal and prevent interference or ground loop issues.

Isolation Barrier

Input circuit Amplifier Output Circuit

Input Circuit:
Input circuit is designed to take a DC signal as an input.
Isolation Barrier:
Use an optocoupler or a transformer to provide electrical isolation between the input and output
circuits.
Amplification Circuit:
The amplification circuit is used to amplify the isolated DC signal.
Output Circuit:
The output circuit provide an isolated amplified DC signal as output.
AC CARRIER AMPLIFIER
• A direct-current amplifier
• The dc input signal is filtered by a low pass filter, then used to modulate a carrier so it
can be amplified conventionally as an alternating-current signal;
• The amplified dc output is obtained by rectifying and filtering the rectified carrier
signal.

1. Carrier Oscillator:
used to energize the transducer with an alternating carrier voltage.
2. Strain gauge transducer:
-The information signal from the body electrodes reaches the transducer where it is
amplitude modulated using carrier signal from the carrier oscillator.
-The transducer changes the amplitude of carrier signal with respect to the physiological
variable being measured.
-The output of transducer is amplitude modulated signal.
3. Amplifier:
-Amplifier used is Multistage Capacitance coupled Amplifier
-The modulated signal from the transducer is given to this amplifier.
-The first stage produces amplification of AM signal.
 -Second stage responds to signal frequency components of carrier signal only.
-Further amplified in the third stage.
4. Rectifier
-Output from the amplifier is converted into unidirectional signal using a rectifier.
5. Phase sensitive Detector
-The signal is demodulated and extracts the amplified information signal.
6. Direct Writing Recorder
-The voltage produced by the detector stage is then fed to the driver stage of the recording
system.
Features of carrier amplifier
• Used to obtain zero frequency response of dc amplifier.
• It reduces wideband noise and increases the signal-to noise ratio.
• This permits the signal to be recovered from its noisy background.
Carrier amplifiers can be used with a resistance strain gauge transducer such as a
semiconductor strain gauge.
When used with pressure gauges, a calibration control is provided on the carrier amplifier.
Lock-in amplifier
Lock-in amplifier is a useful version of the carrier technique designed for the measurement of
low-level signals buried in noise.
POWER LINE INTERFERENCE
Power line interference, also referred to as electrical interference or electrical noise, is
a phenomenon where unwanted electrical signals disrupt the normal functioning of electronic
devices and communication systems. These unwanted signals can originate from a variety of
sources, such as power lines, electronic equipment, radio frequency (RF) radiation, and other
electromagnetic sources.
The powerline interference represents a common noise source in the ECG and other
physiologic signals recorded from the body surface. Depending on the country region, such
noise is characterized by a 50 or 60 Hz sinusoidal interference, possibly accompanied by
harmonics. Powerline interference (50 or 60 Hz noise from mains supply) can be removed by
using a notch filter of 50 or 60 Hz cut-off frequency.
There are several types of power line interference:
Electromagnetic Interference (EMI): This type of interference occurs when electromagnetic
fields generated by power lines or electronic devices induce unwanted currents or voltages in
nearby conductors. These induced currents can disrupt the operation of sensitive equipment.
Radio Frequency Interference (RFI): RFI is caused by radio frequency signals emitted by
various sources, including wireless communication devices, broadcast stations, and radar
systems. These signals can interfere with electronic devices that are not properly shielded
against such frequencies.
Harmonic Distortion: Power lines can sometimes carry harmonics, which are multiples of the
fundamental frequency (usually 50 or 60 Hz). These harmonics can lead to voltage and current
distortions that affect the performance of connected devices.
RIGHT LEG DRIVEN ECG AMPLIFIER
A Driven Right Leg circuit or DRL circuit, also known as Right Leg Driving technique,
is an electric circuit that is often added to biological signal amplifiers to reduce common-mode
interference. Biological signal amplifiers such as ECG (electrocardiogram) EEG
(electroencephalogram) or EMG circuits measure very small electrical signals emitted by the
body, often as small as several micro-volts (millionths of a volt).
However, the patient's body can also act as an antenna which picks up electromagnetic
interference, especially 50/60 Hz noise from electrical power lines. This interference can
obscure the biological signals, making them very hard to measure. Right leg driver circuitry is
used to eliminate interference noise by actively cancelling the interference.

In most modern electrocardiographic systems, the patient is not grounded at all. Instead,
the right-leg electrode is connected to the output of an auxiliary op amp. The common-mode
voltage on the body is sensed by the two averaging resistors Ra, inverted, amplified, and fed
back to the right leg. This negative feedback drives the common-mode voltage to low value.
The body's displacement current flows not to ground but rather to the op-amp output circuit.
This reduces the interference as far as the ECG amplifier is concerned and effectively grounds
the patient.
 The circuit can also provide some electric safety. If an abnormally high voltage should
appear between the patient and ground as a result of electric leakage or other cause, the
auxiliary op amp in saturates. This effectively ungrounds the patient, because the amplifier can
no longer drive the right leg. Now the parallel resistances Rf, and Ro, are between the patient
and ground. They can be several megohms in value-large enough to limit the current. These
resistances do not protect the patient, however, because 120 V on the patient would break down
the op-amp transistors of the ECG amplifier, and large currents would flow to ground.
BAND PASS FILTERING.
A band pass filter (also known as a BPF or pass band filter) is defined as a device that allows
frequencies within a specific frequency range and rejects (attenuates) frequencies outside that
range.

The low pass filter is used to isolate the signals which have frequencies higher than the cutoff
frequency. Similarly, the high pass filter is used to isolate the signals which have frequencies
lower than the cutoff frequency.

By the cascade connection of high pass and low pass filter makes another filter, which allows
the signal with specific frequency range or band and attenuate the signals which frequencies
are outside of this band. This type of filter is known as Band Pass Filter.

Band Pass Filter Types

1. Active Band Pass Filter

Block Diagram of Active Band Pass Filter

The circuit diagram of Active Band Pass Filter is divided into three parts. The first part is for
a high pass filter. Then the op-amp is used for the amplification. The last part of the circuit is
the low pass filter.

Circuit Diagram of Active Band Pass Filter

 2. Passive Band Pass Filter
The passive filter used only passive components like resistors, capacitors, and inductors.
Therefore, the passive band pass filter is also used passive components and it does not use the
op-amp for amplification. So, like an active band pass filter, the amplification part is not present
in a passive band pass filter. The passive band pass filter is a combination of passive high pass
and passive low pass filters.

Circuit Diagram of Passive Band Pass Filter

3. Wide Band Pass Filter
The circuit diagram of this filter is as shown in the below figure where the first half is for active
high pass filter and the second half is for active low pass filter. It is easy to design the circuit
for a wide range of bandwidth.

Circuit Diagram of Wide band Pass Filter

4. Narrow Band Pass Filter
The bandwidth of this filter is narrow. Therefore, it allows the signal with a small range
of frequencies. It has multiple feedback. This band pass filter uses only one op-amp. In this
band pass filter, the op-amp is used in non-inverting mode.
 Circuit Diagram of Narrow Band Pass Filter
Below figure differentiate the frequency response between wide pass and narrow pass filter.
UNIT IV MEASUREMENT OF NON-ELECTRICAL PARAMETERS
Temperature, respiration rate and pulse rate measurements. Blood pressure: indirect methods -
Auscultatory methods, oscillometric method, direct methods: electronic manometer, Pressure
amplifiers, Systolic, diastolic, mean detector circuit, Blood flow and cardiac output
measurement. Indicator dilution, thermal dilution and dye dilution methods, Electromagnetic
and ultrasound blood flow measurement.

TEMPERATURE, RESPIRATION RATE AND PULSE RATE MEASUREMENTS.

Temperature measurements

Temperature measurement refers to the process of quantifying the degree of the human body.
Temperature is typically measured using various instruments and sensors, such as
thermometers, thermocouples, resistance temperature detectors (RTDs), and infrared
thermometers. These devices detect changes in physical properties, such as expansion or
electrical resistance, that are influenced by temperature.

Principle

Temperature measurement relies on the principle that temperature affects physical properties
of materials, such as their volume, electrical resistance, or thermal radiation. Different types of
thermometers and sensors are used to detect these changes and convert them into temperature
values, which are typically expressed in degrees Celsius (°C) or Fahrenheit (°F).

Normal and abnormal temperature:

Normal Range: The normal body temperature for humans varies slightly depending on the
method of measurement and the time of day. Generally, the accepted normal range for oral
temperature is around 36.1°C to 37.2°C (97.0°F to 98.9°F), while rectal temperature falls
within a slightly higher range due to the body's core temperature.

Abnormal Range: Abnormal body temperatures can indicate various health conditions:

Fever: A body temperature above the normal range is typically considered a fever. In adults,
this is often defined as a temperature above 38°C (100.4°F). Fever is often a sign of infection
or inflammation.

Hypothermia: When the body temperature drops below the normal range, it's known as
hypothermia. This can occur in cold weather or due to prolonged exposure to cold water and
can be life-threatening if severe.

Hyperthermia: Hyperthermia refers to a very high body temperature, often caused by heat
stroke or heat exhaustion. It can lead to heat-related illnesses and should be treated promptly.

PULSE RATE MEASUREMENTS

Pulse rate measurements

Pulse rate measurements, also known as heart rate measurements, involve counting the number
of times a person's heart beats per minute. This is typically done by feeling the pulse at various
locations on the body or by using medical devices like pulse oximeters or heart rate monitors.
A normal resting pulse rate for adults is usually between 60 and 100 beats per minute, but it
can vary based on factors such as age, fitness level, and overall health. Regularly monitoring
pulse rate can provide valuable information about cardiovascular health and physical fitness.

Principle: Pulse rate is typically measured by palpating (feeling) the pulse at specific
anatomical locations, such as the wrist (radial artery), neck (carotid artery), or chest (apical
pulse). The principle involves counting the rhythmic pulsations generated by the heart's
contractions over a 60-second interval or a shorter time frame (e.g., 15 seconds, then
multiplying by 4) to determine the heart rate in beats per minute (BPM).

Normal Range: The normal resting pulse rate for adults typically falls between 60 and 100
BPM. However, normal ranges can vary based on factors such as age, fitness level, and overall
health. Athletes, for example, may have lower resting heart rates due to their conditioning.

Abnormal Range: Abnormal pulse rates can indicate various health conditions:

Bradycardia: A pulse rate below 60 BPM at rest may suggest bradycardia, which could be
caused by heart conduction issues, medication side effects, or other underlying health
problems.

Tachycardia: A pulse rate above 100 BPM at rest may indicate tachycardia, which can result
from stress, fever, dehydration, anemia, or cardiac issues.

Arrhythmia: Irregular pulse rhythms, such as atrial fibrillation, can lead to abnormal pulse
rates. In atrial fibrillation, the heart's electrical signals are disorganized, causing an irregular
pulse.

RESPIRATORY MEASUREMENT

Respiratory measurement
Respiratory measurement is the process of quantifying various aspects of breathing and lung
function in individuals. This includes parameters such as respiratory rate, tidal volume, minute
ventilation, and lung capacity. Respiratory measurements are crucial for assessing respiratory
health, diagnosing conditions, and monitoring treatment.

Principles:
Respiratory measurements are based on the principles of airflow and gas exchange in the
respiratory system. Various instruments and techniques are employed to capture data related to
how air moves in and out of the lungs, as well as the exchange of oxygen and carbon dioxide
during respiration.

Normal and Abnormal Range (Respiratory Rate):

Normal Respiratory Rate: In adults, the normal resting respiratory rate typically ranges from
12 to 20 breaths per minute.

Abnormal Respiratory Rate: A respiratory rate outside this range, either too high (tachypnea)
or too low (bradypnea), can be indicative of underlying health issues. For example, a rate
consistently above 20 breaths per minute (tachypnea) or below 12 breaths per minute
(bradypnea) may warrant further medical evaluation.

BLOOD PRESSURE: INDIRECT METHODS AND DIRECT METHODS

Blood pressure measurements

The blood pressure measurement is typically expressed in two numbers: systolic and diastolic
pressure. The systolic pressure is the higher of the two and represents the pressure in the arteries
when the heart contracts (beats), while the diastolic pressure is the lower number and represents
the pressure in the arteries when the heart is at rest between beats.

Normal Range:
Blood pressure is measured in millimeters of mercury (mm Hg). The normal range for blood
pressure in adults is typically considered to be:
Systolic Pressure: Less than 120 mm Hg
Diastolic Pressure: Less than 80 mm Hg

Abnormal Range:
Abnormal blood pressure readings can indicate various health conditions. Abnormal blood
pressure can be categorized as:

Hypertension (High Blood Pressure):

Stage 1 Hypertension: Systolic 130-139 mm Hg or diastolic 80-89 mm Hg

Stage 2 Hypertension: Systolic 140 mm Hg or higher or diastolic 90 mm Hg or higher

Hypotension (Low Blood Pressure):

Hypotension is less common but can be defined as having a systolic blood pressure below 90
mm Hg and a diastolic pressure below 60 mm Hg. It can result from various factors, including
dehydration, medication side effects, or underlying medical conditions.

INDIRECT METHODS - AUSCULTATORY METHODS AND OSCILLOMETRIC

METHOD.

1. Indirect Blood Pressure Measurement:

Indirect methods of measuring blood pressure are the most common and non-invasive
techniques. They are widely used in clinical settings and for routine monitoring. There are two
primary indirect methods such as auscultatory methods and oscillometric method.

Oscillometric Blood Pressure Measurements:

Oscillometric blood pressure measurement is a common method for non-invasive monitoring

of blood pressure. It relies on the principle of detecting oscillations (vibrations) in the arterial
wall caused by the pulsatile flow of blood through the arteries. This technique is frequently
used in automated electronic blood pressure monitors and is well-suited for self-monitoring
and clinical use.
Principle:
The oscillometric method records and evaluates the oscillation of arteries with the help of
pressure transducer. The oscillometric method is based on the observation that when a blood
pressure cuff is inflated and then slowly deflated, the pressure within the cuff oscillates as it
approaches the systolic and diastolic pressures. These oscillations are caused by the periodic
expansion and contraction of the artery under the cuff, which corresponds to the pulsatile flow
of blood.

Components: Typical components of an oscillometric blood pressure monitor include:

Cuff: This is wrapped around the patient's upper arm or wrist.
Pressure Sensor: It measures the pressure within the cuff and detects the oscillations.
Pump: In automated monitors, a pump is used to inflate the cuff.
Display: The monitor displays the blood pressure readings, typically as systolic/diastolic
pressure in millimetres of mercury (mm Hg).

Working:
The working principle of oscillometric blood pressure measurement involves several steps:
Inflation: The blood pressure cuff is wrapped around the upper arm (or another suitable
location). It is inflated to a pressure level significantly above the expected systolic blood
pressure.
Deflation: The cuff is slowly and gradually deflated.
Detection of Oscillations: As the cuff pressure decreases, the arterial blood flow beneath the
cuff is partially constricted and then released. This leads to vibrations in the arterial wall,
causing the pressure within the cuff to oscillate.
Amplitude Analysis: The electronic blood pressure monitor detects these oscillations and
analyzes their amplitude (size) and frequency.
Determination of Blood Pressure: The monitor uses algorithms to determine the systolic and
diastolic blood pressures based on the characteristics of the oscillations.
 Figure - A) Blood pressure measurement by oscillometric methods

Auscultatory Blood Pressure Measurements:

Auscultatory blood pressure measurement is a traditional method for measuring blood pressure
using a stethoscope and a manual or mercury sphygmomanometer. This method relies on the
principle of listening for specific sounds (Korotkoff sounds) in the arteries while gradually
releasing pressure from the cuff. It has been widely used in clinical settings for many years.

Principle:
The auscultatory method is based on the observation that when a blood pressure cuff is inflated
and then slowly deflated, the pressure within the cuff reaches levels where it partially obstructs
and then releases blood flow through the brachial artery. These changes in blood flow result in
specific sounds called Korotkoff sounds. The first Korotkoff sound corresponds to the systolic
blood pressure, and the point where the sounds disappear corresponds to the diastolic blood
pressure.

Components:
Components of an auscultatory blood pressure measurement setup typically include:
Blood Pressure Cuff: This is wrapped around the patient's upper arm, and it contains an
inflatable bladder to apply pressure to the artery.
Sphygmomanometer: This is the pressure-measuring device, which can be a manual aneroid
sphygmomanometer or a mercury sphygmomanometer.
Stethoscope: A stethoscope with a bell or diaphragm is used by the healthcare provider to
listen to the Korotkoff sounds.

Working:
The working principle of auscultatory blood pressure measurement involves several steps:
Inflation: The blood pressure cuff is wrapped around the upper arm, typically at heart level. It
is inflated to a pressure level significantly above the expected systolic blood pressure.
Deflation: The cuff is slowly and gradually deflated at a controlled rate, usually about 2-3 mm
Hg per heartbeat.
Detection of Korotkoff Sounds: Uses a stethoscope to listen for Korotkoff sounds over the
brachial artery (inside of the elbow). These sounds are heard as the pressure in the cuff falls.
Identification of Phases: There are five recognized phases of Korotkoff sounds. The first
sound (Phase I) corresponds to the onset of the clear, rhythmic tapping sounds and indicates
the systolic pressure. The fifth sound (Phase V) corresponds to the point where the sounds
disappear or become muffled, indicating the diastolic pressure.
Determination of Blood Pressure: The healthcare provider records the systolic and diastolic
pressures based on the appearance and disappearance of Korotkoff sounds.
 Figure - B) Blood pressure measurement by auscultation methods

DIRECT METHODS: ELECTRONIC MANOMETER, PRESSURE AMPLIFIERS,

SYSTOLIC, DIASTOLIC, MEAN DETECTOR CIRCUIT.

Direct Methods: Electronic Manometer

An electronic pressure transducer can be connected to the patient through a thin piece of tubing
called a catheter. The catheter is introduced into the vessel through a thin, hollow tube called a
cannula. The transducer's pressure diaphragm is coupled to the patient's bloodstream by a
column of saline solution that fills the catheter.

The catheter to the patient must be placed inside a peripheral artery. There are two general
methods for inserting the catheter: percutaneous puncture and arterial cutdown. The
percutaneous methods involve puncturing the skin over an artery and then using a needle and
catheter assembly to insert the catheter into the artery. When the catheter is in place, the needle
is withdrawn. The arterial cutdown method is a surgical procedure in which the tissue overlying
the artery is cut and laid out of the way, revealing the artery. A puncture is then made, and a
catheter is put into place.

Figure shows a typical apparatus used for measuring blood pressure with an electronic
transducer. The transducer is mounted to a pole or support at bedside. There are two ports into
the transducer's pressure dome; one is "deadheaded" and the other is connected to a hydraulic
fitting called a three-way stopcock. One terminal of the stopcock goes to the catheter from the
patient, while the other goes to a small syringe that is used to administer medications directly
to the patient through the catheter, or to withdraw arterial blood samples for laboratory analysis.

• Catheter tip probe sensor mounted at the tip of the probe. Pressure exerted on the tip is
converted to the corresponding electrical signal.
• In fluid filled catheter type. Pressure exerted on the fluid filled column is transmitted to
external transducer. This transducer converts pressure into electrical signal.
• Here fluid filled catheter is used. Before inserting catheter into blood vessel, fluid filled
system should be completely flushed out. Usually, sterile saline is used for this purpose.
Because blood clotting is avoided
Pressure Amplifiers

A blood pressure amplifier, also known as a blood pressure transducer or pressure sensor, is a
complex device with several key components that work together to accurately measure and
amplify the blood pressure signal.

1. Pressure Sensor:

The pressure sensor is the primary component responsible for directly sensing the pressure
changes within the blood pressure cuff. It typically consists of a diaphragm or membrane that
deforms when subjected to pressure. This deformation generates a mechanical signal that
corresponds to the applied pressure.

2. Transducer:

The mechanical signal generated by the pressure sensor needs to be converted into an electrical
signal that can be processed and amplified. This is the role of the transducer. There are various
types of transducers used in blood pressure amplifiers, including strain gauges, piezoelectric
sensors, and capacitive sensors.

3. Amplification Circuitry:

The electrical signal produced by the transducer is usually very weak and needs to be amplified
to make it suitable for measurement and display. Amplification circuitry consists of electronic
components such as operational amplifiers (op-amps) and amplification stages that increase the
signal's strength while maintaining accuracy.

4. Signal Conditioning:

The raw amplified signal may contain noise and artifacts that need to be filtered and removed
for accurate blood pressure measurement. Signal conditioning circuitry includes filtering and
analog signal processing to enhance the quality of the blood pressure signal.

5. Calibration Components:

Calibration is a critical aspect of blood pressure amplifiers to ensure accurate readings.

Calibration components are used to adjust and calibrate the amplifier so that the output
corresponds correctly to actual blood pressure values. This may involve setting the zero-
pressure reference and the gain of the amplifier.

6. Display and Output Components:

The processed and calibrated blood pressure signal is typically displayed on a monitor or
recorded for further analysis. Display and output components include digital displays, analog
outputs, or connections to data acquisition systems. These components allow healthcare
professionals to read and interpret the patient's blood pressure.

Systolic, Diastolic and Mean Detector Circuit.

The pressure amplifier produces an analog wave- form with a peak amplitude representing the
systolic pressure, and a minimum, or "valley," that represents the diastolic pressure. Additional
circuitry is required that recognizes these points, and from them produces a steady dc voltage
or current that can be used to drive a display meter.

1. This circuit operates from a two-phase clock created by flip-flop F̅̅̅̅

F1. The Q and not
Q (Q) outputs are complementary, so one will be high when the other is low.

2. Switches S1 and S4 are turned on when the Q is high, and switches S2 and S1 are turned
on when the 𝑄̅ is high.

3. The analog waveform from the output of the pressure amplifier is applied
simultaneously (i.c., in parallel) to the inputs of A1 and A2 The waveform, therefore,
appears simultaneously at the outputs of A1 and A2.

Figure 1 – Systolic detector circuit and diastolic detector circuit

4. During period T1, the Q output of F̅̅̅̅

F1 is high, so switches S1 and S4 are closed. When
S4 is closed, capacitor C2 is discharged, so it has no effect on the output. Closing switch
 S1 allows the signal appearing at the output of A1 to charge capacitor C1 to the peak
voltage, representing the systolic pressure.

Figure - Timing diagram

5. The voltage across capacitor C1 forward biases diode D3, which then conducts and
applies the voltage to the input of amplifier A3. The output of amplifier A3 goes to the
systolic output meter.

6. The situation reverses during the time T2: The 𝑄̅ of FF1 becomes high and the Q is low.
This turns on switches S2 and S3 and turns off S1 and S4. Closing S2 causes capacitor
C2 to rapidly charge to the peak voltage of the input signal. This occurs within less than
a second in most cases.

7. Switch S3 closes to allow capacitor C1 to discharge slowly through resistor R3. The
charge on C2 will reach peak before any appreciable decay of the C1 charge takes place.
In some models each charge switch is operated directly from FF1. while the discharge
switch is operated by the same signal after it has passed through an RC network delay
line. This network ensures that one capacitor is fully charged before the other begins to
decay.

8. The voltage at the output of Figure 1 represents the peak of the waveform, which
corresponds to the systolic pressure. The same circuit may be used to detect the diastolic
voltage by inverting the waveform so that the diastolic feature becomes the peak.

The MAP is found by taking the time average of the pressure waveform. Most pressure
monitors use a simple RC integrator with buffering amplifiers rather than regular operational
amplifier integrators.
 Figure 2 – MAP detector

BLOOD FLOW AND CARDIAC OUTPUT MEASUREMENT.

Blood Flow Measurement

• Blood flow measurement refers to the quantification of the rate of blood circulation in
a particular blood vessel or organ, typically expressed in units like milliliters per
minute.
Principle:
• Blood flow measurement is based on the principle of quantifying the volume of blood
passing through a particular point in the circulatory system over a specified time.
• Several methods are used to achieve this, including Doppler ultrasound, magnetic
resonance imaging (MRI), and thermodilution.
Types:
• Doppler Ultrasound: This method uses the Doppler effect to assess blood flow
velocity. It can measure both direction and speed of blood flow.
• Thermodilution: Often used in cardiac catheterization, this method involves
injecting a known volume of cold saline into the bloodstream and measuring how
it dilutes as it mixes with warm blood.
• Electromagnetic Flowmetry: This measures blood flow by detecting changes in
electromagnetic properties as blood moves through a vessel.
 • Laser Doppler Flowmetry: This technique uses laser light to assess blood flow in
small vessels near the skin's surface.

Normal Range:
• Normal blood flow values can vary depending on the location and size of the vessel or
organ being measured. However, typical blood flow rates for a healthy adult may range
from 70 to 100 milliliters per minute per 100 grams of tissue for many organs.
Abnormal Conditions:
• Abnormal blood flow can be indicative of various medical conditions:
• Hypoperfusion: Insufficient blood flow, which can lead to tissue damage or organ
dysfunction.
• Hyperperfusion: Excessive blood flow, which may indicate inflammation or other
pathologies.

Blood Flowmeter Method

• Electromagnetic Blood Flowmeter Method
• Ultrasonic Blood Flowmeter Method

ELECTROMAGNETIC BLOOD FLOWMETER

• An electromagnetic blood flow meter is a medical device used to measure the velocity
and volume of blood flow in a patient's blood vessels.
• This type of meter is widely used to measure pulsatile blood flow.

Principle:

• It operates on the principle of electro-magnetic induction, which is based on Faraday's

law of electromagnetic induction.

• When a blood flows through a magnetic field, it generates a voltage that is directly
proportional to the velocity of the blood.

Block Diagram:
 • It uses an oscillator of low frequency (up to 400 Hz) to drive the electromagnet which
is placed in such a way that the magnetic field is perpendicular to the direction of blood
flow.
• An EMF is induced across the blood vessel.
• In order measure this, a set of electrodes are placed across the blood vessel mutually
perpendicular to both magnetic field and direction of blood flow.

• The use of a gated detector makes the polarity of the output signal reverse when the
flow direction reverses.
• The EMF induced across the blood vessel will be proportional to the velocity of blood,
Lumen probes with varying diameters are used for the accuracy of measurement.
• The output of electromagnetic blood flow meter, which is in micro volts, is enhanced
by the amplifier.
• For the average blood flow rate, usually use LPFS after the amplifier.
• Amplifier drift and electrode polarization may cause some problems with
electromagnetic type flow meters.

ULTRASOUND BLOOD FLOW MEASUREMENT

• An ultrasonic flow meter is a type of flow meter that measures the velocity of a fluid
with ultrasound to calculate volume flow.
• The blood cells in the fluid reflects the ultrasound signal with a shift in the ultrasonic
frequency due to its movement.
• Using ultrasonic transducers, the flow meter can measure the average velocity along
the path of an emitted beam of ultrasound, by averaging the difference in measured
transit time between the pulses of ultrasound propagating into and against the direction
of the flow or by measuring the frequency shift from the Doppler effect.
 • A beam of ultrasonic energy is used to measure the velocity of flowing blood.
• Lead zirconate titanate is a crystal that has the highest conversion efficiency.

Two types:
• Transit time flow meter
• Doppler type

Transit time ultrasonic flow meter

• The pulsed beam is directed through a blood vessel at a shallow angle and its transit
time is measured.
• The transit time is shortened when the blood flows in the same direction as the
transmitted energy.
• The transit time is lengthened otherwise.

Block diagram:
 • A piezoelectric crystal emits a brief pulse of ultrasound which propagates diagonally
across the blood vessel.
• If the flow is in the same direction as the pulse, then the pulse reaches a receiving
crystal situated on the opposite side wall of the blood vessel.
• Appropriate electronics can convert the change in transit time to velocity.
• In the figure, 'D' is the distance travelled by sound waves along the downstream of
the blood flow and it is the distance between the transmitter and receiver of ultrasonic
waves.
• Here is the angle between the direction of flow and the central axis of the ultrasonic
beam, C is the ultrasonic velocity in blood.
• First the switch selector closes S1 and opens S2. This connects the R.F. generator to
crystal 1 and the receiver to crystal 2 and upstream transit time is measured.
• Subsequently the switch selector opens S1 and close S2.
• This connects the R.F. generator to crystal 2 and the receiver to crystal 1 and measure
downstream transmit time.
• Thus, the blood flow velocity can be measured by determining the difference be
upstream and downstream transit time.

Doppler Type Ultrasonic flow meters

• Based on the Doppler principle

• A transducer sends an ultrasonic beam with a frequency F into the flowing blood.
• A small part of the transmitted energy is scattered back and is received by a second
transducer arranged opposite the first one.
• The reflected signal has a different frequency F+FD or F-FD due to Doppler effect.

Block diagram:
 • An oscillator, operating at a frequency of several megahertz, excites a piezoelectric
transducer (usually made of barium titanate).
• This transducer is coupled to the wall of an exposed blood vessel and sends an
ultrasonic beam with a frequency F into the flowing blood.
• A small part of the transmitted energy is scattered back and is received by a second
transducer arranged opposite the first one.
• Because the scattering occurs mainly as a result of the moving blood cells, the reflected
signal has a different frequency due to the Doppler effect.
• Its frequency is either F + FD or F - FD, depending on the direction of the flow.
• The Doppler component FD is directly proportional to the velocity of the flowing blood.
• A fraction of the transmitted ultrasonic energy, however, reaches the second transducer
directly, with the frequency being unchanged.
• After amplification of the composite signal, the Doppler frequency can be obtained at
the output of a detector as the difference between the direct and the scattered signal
components.

CARDIAC OUTPUT MEASUREMENT

Cardiac Output Measurement

Cardiac output refers to the volume of blood that the heart pumps per minute, usually measured
in liters per minute (L/min).

Principle:
• Cardiac output is calculated using various methods that involve measuring blood flow,
either directly or indirectly.
• The principle behind these methods is to determine the volume of blood leaving the
heart per unit of time.

Normal Range:

• The normal range for cardiac output can vary depending on factors such as age, sex,
and body size.
• In adults, the typical resting cardiac output ranges from 4.5 to 6.0 liters per minute.
Abnormal Conditions:
• Abnormal cardiac output can indicate various cardiovascular and systemic health
issues.
• Low cardiac output (cardiac output insufficient to meet the body's demands) can occur
in conditions like heart failure, cardiogenic shock, or severe dehydration.
• High cardiac output (excessive blood flow from the heart) can be seen in conditions
like hyperthyroidism, anemia, or arteriovenous fistulas.

CARDIAC OUTPUT METHODS

• Indicator Dilution
• Thermal Dilution
• Dye Dilution Methods

INDICATOR DILUTION
• To calculate cardiac output by the “indicator dilution method,” a small amount of
indicator, such as a dye, is injected into a large systemic vein or, ideally, into the right
atrium.
• This indicator travels quickly through the right side of the heart, then through the lungs’
blood vessels, the left side of the heart, and eventually through the systemic arterial
system.
• The concentration of the dye is measured as it passes through one of the peripheral
arteries, resulting in the curve portrayed in image.
• In each of these cases, 5 milligrams of Cardiogreen dye were injected at the start of the
experiment.

• In the top recording, none of the dye entered the arterial tree until about 3 seconds after
the injection, but then the arterial concentration of the dye rapidly increased to a
maximum in about 6 to 7 seconds.
• After that, the concentration fell quickly, but some of the dye had already circulated all
the way through some of the peripheral systemic vessels and returned through the heart
for a second time.
• As a result, the dye concentration in the artery started to increase once more. For the
purpose of measurement, it is important to extrapolate the early down-slope of the curve
to the zero point, as shown by the dashed portion of each curve.
 • The amplitude of this portion depends upon the quantity of injected indicator and on
the total quantity of the circulating blood.

Cardiac output = indicator dose / area under the concentration-time curve

THERMAL DILUTION

• Therefore now-a-days thermo dilution method is adopted to measure cardiac output.

• A bolus of about 10 millilitres of 5% Dextrose in water at room temperature is injected
as a Thermal indicator into the right atrium.
• After mixing, it is detected in the pulmonary artery by means of a thermistor mounted
at the tip of a miniature catheter probe.
• The temperature difference between the injectate temperature and the circulating blood
temperature in the pulmonary artery is measured.
• The reduction in temperature is integrated with respect to time.
• After applying proper corrections, a meter reads the cardiac output.
• Assuming that the thermo indicator mixes thoroughly with the blood and negligible
heat flow across the blood vessel wall, the heat injected is equal to heat conducted.

• A thermal indicator of known volume introduced into either the right or left atrium will
produce a resultant temperature change in the pulmonary artery or in the aorta
respectively, the integral of which is inversely proportional to the cardiac output.

Block diagram:
 • The nonlinearity of the thermistor is removed by connecting a parallel resistor with that
and hence a linear relation between temperature and resistance of the thermistor can be
obtained.
• Based on this fact, the linearising amplifier works.
• The integrator delivers the value of the integral of the blood temperature change over a
given measurement time.
• By feeding data about the density and specific heat of blood and thermal indicator and
volume of thermal indicator injected, the computer can deliver the cardiac output in
liters/minute.

DYE DILUTION METHODS

• The dye dilution method is a technique used to measure cardiac output, which is the
volume of blood the heart pumps per minute.
1. A known quantity of a dye (often indocyanine green or a similar substance) is injected
into a large central vein, such as the right atrium of the heart.
2. The dye mixes with the blood and travels through the circulatory system.
3. As the dyed blood passes through an artery, samples are taken at regular intervals.
4. The concentration of dye in these arterial blood samples is measured, often using a
photometer which measures light absorbance.

Cardiac Output=Mean concentration of dye in arterial blood samples/Quantity of dye injected

 • The photometric part consists of a source of radiation and a photocell and an
arrangement for holding the disposable polyethylene tube constituting the cuvette.
• An interference filter with a peak transmission of 805 nm is used to permit only
infrared radiation to be transmitted.
• This wavelength is the isobestic wavelength for haemoglobin at various levels of
oxygen saturation.
• In order to avoid the formation of bubbles, the cuvette tubing should be flushed with a
solution of silicone in ether.
• A flow rate of 40 ml/min is preferred in order to get as short a response time as possible
for the sampling catheter.
• The sampling syringe has a volume of 50 ml/min.
• The output of the photocell is connected to a low drift amplifier. It has a high input
impedance and low output impedance.
• The amplification is directly proportional to the resistance value of the potentiometer
R.
• A potentiometric recorder records the amplifier signal on a 200 mm wide recording
paper and a paper speed of 10 mm/s.

RESPIRATION METHOD

1. Displacement method
2. Thermistor method
3. Impedance pneumography
4. Apnea detectors
1. Displacement Method
In this method the transducer is hold by an elastic band which goes around the chest.
The respiratory movements results in a corresponding resistance changes of the strain gauge.
It is connected as one arm of a wheatstone bridge circuit. Its output varies with chest expansion.
This output corresponds to the respiration activity.
2. Thermistor Method
Generally, there is a temperature difference between inspired and expired air. This temperature
is sensed by placing thermistor in front of nostrils. Thermistor is placed by using suitable stand.
The thermistor is connected with the bridge circuit. So, unbalance signal is amplified to get the
respiratory activity.
3. Impedance Pneumography
This is the indirect method of measurement. Impedance pneumography is based on the
fact that, the a.c impedance across the chest of a patient changes as respiration occurs. 50-
50KHz a.c signal is produced by oscillator circuit and is given to the chest of the patient through
electrodes.
The signal voltage applied to the amplifier (Differential amplifier) block is the voltage
drop across the resistance.

V = I(R+ R)

Where V= Output voltage (V)

I= Current through the chest (A)
R= chest impedance without respiration (R)
R= change of chest impedance due to respiration (Q)

The output of the amplifier is given to demodulator and filter block. Hence low pass filter is
used to remove the residual carrier signal. The output of the impedance pneumograph contains
respirating rate data.
4. Apnea Detectors
Apnea is the stopping of breathing. It leads to the arrest of the circulation. It can be occurred at
the conditions like head injury, drug overdose, etc. It can also occur in premature babies during
the first week of life because of their immature nervous system. If apnea persists for a
prolonged period, then brain functions can be severely damaged. So, apnea patients are closely
monitored. Apnea monitor is used to watch the apnea patient’s respiration rate. Apnea monitor
gives audio signals and visual signals, when no inspiration occurs for a particular period of
time. Input from the sensor is connected with the amplifier circuit having high input impedance.
The output of the amplifier circuit is connected with motion and respiration channel blocks.
Motion channel block differentiates motion and the respiration based on the frequency. The
frequency below 1.5 Hz is identified as respiration. The frequency above 1.5 Hz is identified
as motion. High frequency signal above the threshold is sensed by positive detector.
The frequency below the threshold is sensed by negative detector. The output of the motion
channel is connected with comparator circuit. It compares the amplitude of motion and
respiration. Based on the output corresponding lamp will glow. In the respiration channel, low
pass filter is used to remove high frequency signal. If there is no respiration, then Schmitt
trigger circuit gives the output to switch on the alarm.
Apnea period selector circuits contain low frequency alarm oscillator and tone oscillator, and
audio amplifier. Apnea period selector circuit drives the alarm circuit. The output of alarm
circuit is connected with the speaker. So, where there is no respiration for a period of 10 or 20
sec, then audio signal through the speaker and visual signal through the flash light is delivered.
PULSE METHOD
When heart muscle contracts, blood is ejected from the ventricles and a pulse of pressure
is transmitted through the circulatory system. This pulse can be measured at various points. We
can sense the pulse by placing our finger-tip over the radial artery in the wrist. This pulse travels
at the speed of 5 to 15m per second. Photoelectric method is commonly employed to measure
the pulse.
Types:
Photoelectric method consists of two types
· Transmittance method
· Reflectance method
1. TRANSMITTANCE METHOD OF PULSE MEASUREMENT
LED and photoresistor are used in this method. These are mounted in a enclosure that fits over
the tip of the finger. Light is produced by the LED. The same light is passed through the finger.
For each heart pulse, blood is forced to the extremities and the amount of blood in the finger is
increased. So optical density is changed. So, the light transmitted through the finger is
decreased. This light is received by the photo resistor. This photo resistor is connecte with the
part of voltage divider circuit. The voltage produced by this circuit is directly proportional to
the amount of blood flow in the figure. The output ius recorded by using strip chart recorder.

2. REFLECTANCE METHOD
In the reflectance method, LED is placed adjacent to the photoresistor. LED emits the light.
This light is reflected form the skin and the tissues fall on the photo resistor. The reflected light
varies depending upon the blood flow in the finger. The photo resistor is connected as a part of
the voltage divider circuit. The output obtained is directly proportional to the amount of blood
in the finger. The output can be recorder using strip chart recorder.

TEMPERATURE METHOD
THERMISTORS:
• Thermal+resistor.
• It generally composed of semiconductor material.
• Most thermistor have a negative temperature co efficient of temperature resistance
Principle:
Thermistor circuit are used to detect very small change in temperature. The resistance of
conductor changes with change in the temperature.
Construction:
• They are available in variety of sizes and shapes. it may in form of beads, rods and discs
shaped structure.
• When temperature changes, the resistance of the thermistor changes in a predictable
way.
Two Types of Thermistors
• Negative temperature co efficient (NTC) - when the temperature increases, the
resistance decreases.
• Positive temperature co efficient (PTC) - when the temperature increases the resistance
also increased
THERMOCOUPLE:
• The thermocouple is one of the simplest and most commonly used method of measuring
process temperature.
• It operates based on seeback effect.
• When the heat is applied to junction of two dissimilar metal,
• an emf is generated and it can be measured at another junction (cold junction).
• Two dissimilar metals form and electrical circuit and a current flow as a result of a
generated emf as shown in the fig.
• The current will continue to flow as long as T1 >> T2.
• The emf produced in a function of the difference the temperature of hot and cold.it is
given by,
∈= 𝑎∆𝜃 ∆𝜃 = 𝑑𝑖𝑓𝑓𝑒𝑟𝑎𝑛𝑐𝑒 𝑏𝑒𝑡𝑤𝑒𝑒𝑛 𝑡𝑒𝑚𝑝𝑒𝑟𝑎𝑡𝑢𝑟𝑒 𝑎𝑡 ℎ𝑜𝑡 𝑎𝑛𝑑 𝑐𝑜𝑙𝑑 𝑗𝑢𝑛𝑐𝑡𝑖𝑜𝑛.

Principle:
In thermocouple temperature measuring circuit the emf set up is measured by sending a current
through a moving coil instrument, the deflection being directly proportional to emf. Since emf
in a function of temperature difference ∆𝜃 the instruments can be calibrated to read a
temperature.
UNIT V BIOCHEMICAL MEASUREMENT AND BIOSENSORS
Biochemical sensors - pH, pO2, pCO2, Ion selective Field effect Transistor (ISFET).
Immunologically sensitive FET (IMFET), Blood glucose sensors, Blood gas analyzers,
colorimeter, Sodium Potassium Analyzer, spectrophotometer, blood cell counter, auto analyzer
(simplified schematic description) - Bio Sensors - Principles - amperometric and voltometric
techniques.

BIOCHEMICAL SENSORS - pH, pO2, pCO2,

The pH electrode
The chemical balance of the body is identified by the measurement of pH of blood and other
body fluids. The pH is defined as the logarithm of the reciprocal of the H+ ion concentration.

pH = log10 1/ [H+]

A neutral solution has a pH of 7. When the value of pH of a solution is more than 7 then it is
basic solution When the value of pH is lower than 7, then the solution is acidic solution. Human
blood is slightly basic such that the pH value of venous blood is about 7.35 and for arterial
blood it is about 7.40.

The glass electrode is normally used as a pH electrode. Figure shows the glass electrode
consisting of a spherical bulb 0.5 cm in diameter which provides a thin glass membrane which
permits the passage of only hydrogen ions in the form of H₂O.
Inside the glass bulb silver-silver chloride non polarizable electrode is immersed in
chloride buffer solution usually of pH = 1. The other side of the glass bulb is exposed to the
solution of unknown pH.
The connection to the potential measuring circuit and the solution being tested is
completed through a potassium chloride salt bridge and a calomel reference electrode. Today
the glass electrode and reference electrode are available in the same enclosure. There are many
advantages for glass electrode over hydrogen electrode.
 The glass electrode is independent of oxidation-reduction potentials. It is not necessary
to pass a gas through the solution. Equilibrium is reached rapidly. The electrode can be used in
colored or turbid solutions. It gives a small error when we measure the pH for highly acidic
solutions (near pH = 0) and alkaline solutions (above pH = 9).

The pO2 electrode

The oxygen electrode is a piece of platinum wire embedded in an insulating glass holder
the end of the wire exposed to the electrolyte into which the oxygen from the solution under
measurement allowed to diffuse through the membrane as shown in figure. The bottom of the
vessel containing electrolyte consists of a membrane permeable to oxygen and the top of the
vessel is sealed. Ag-AgCl reference electrode is used.

0.7 volts is applied between the platinum wire and the reference electrode using a
battery. The negative of the battery is connected to the platinum wire through an ammeter.
Reduction of the oxygen takes place at the platinum wire. Due to that, an oxidation - reduction
current is developed and is proportional to the partial pressure of the diffused oxygen. The
oxygen electrode is also used to monitor the partial pressure of oxygen in biological fluids.
Similar to pH electrode, oxygen electrode is available in the integrated version consisting the
platinum cathode and reference electrode in the same enclosure and is called the Clark
electrode.

The pCO2 electrode

This value is measured directly by the CO2 electrode. An increased PCO₂ is often the result of
acute, chronic (or) impending respiratory failure. A decreased PCO2 is the result of
hyperventilation stimulated by a metabolic acidosis (or) hysteria and severe anxiety reactions.
The normal arterial PCO2 is 40 mmHg and the above process is expressed as

PCO2 = Barometric pressure - water vapor pressure x % CO2 100

All modern blood gas analyzers use a PCO₂ electrode. The PCO2 electrode is a modified pH
electrode. There are two major differences between this electrode and the pH electrode.
The first difference is that in this electrode, the blood sample comes in contact with a CO₂
permeable membrane such as Teflon, silicon rubber, rather than a pH sensitive glass as in pH
electrode. The CO from the blood sample diffuses via the CO, permeable (silicon) membrane
into a bicarbonate solution.

ION SELECTIVE FIELD EFFECT TRANSISTOR (ISFET)

Ion selective field effect transistor:
• The potential for low cost, micro miniature sensor that utilize ISFET.
• ISFET employ the same electro chemical principle in their measurement as ion
sensitive electrodes (ISE).
• ISFET is produced by removed of the metal gate region that is normally present on the
FET.
Working:
• The principle is a change of normal FET and they are used in many amplifiers circuits.
• In the ISFET normally input is used as metal gate which replaced by ion sensitive
membrane.
• Hence the ISFET gather in one devise the sensing surface and a single amplifier gives
the high current, low impedance output and it allows.
• There is difference machine for the DH measurement from traditional glass electrode,
the measurement principle is based on the control of current flowing between two
semiconductors, they are drain and source.
• Two semiconductors are placed together to aa third electrode and it behave is like a
gate terminal, the gate terminal is directly contacted to the solution to be measured.

IMMUNOLOGICALLY SENSITIVE FET (IMFET)

Immunologically sensitive field effect transistor (IMFET):
• IMFET is an extension of ISFET.
• ISFET takes advantage of the ion sensitive a chemical sensitive property of FPT.
• IMFET is similar to structure to the ISFET except the solution membrane interface is
polarized than unpolarized that is charged cannot cross the membrane.
• ISFET interact through ion exchange mechanism with the chemical analysis that is
being measured whereas the IMFET operation is based in an antigen antibody reaction.
• An antibody is immobilized on the membrane that is attached to the insulator of a FET.
• In this way the device is used as a antigen sensor. An antibody could be detected in
similar way by immobilizing an antigen on membrane. The IMFET measure charge,
So, in order to be sensed the absorbing species on the membrane must possess a not
electric charge.
BLOOD GLUCOSE SENSORS
Blood glucose sensors
• Blood glucose devices use enzyme-coated test strips that are manufactured with a
precise number of specific enzymes that can only react to one blood sample.
• Because of this, test strips are intended for single use and cannot be reused.
• When inserted into the blood glucose meter and after receiving a blood sample, the test
strip communicates with the glucose meter which calculates the amount of glucose in
the blood and displays the result on the meter’s screen.
•
Components of Blood Glucose Sensors

• Analyte: A substance with chemical constituents that are being identified and
measured. In this instance, glucose is the analyte that the biosensor is designed to detect.
• Bioreceptor: This is a molecule that specifically recognizes the analyte. For the
detection of glucose, specific enzymes are used, which are proteins that facilitate a
chemical reaction. For example, the test strip for a blood glucose test contains the
enzyme that interacts with the analyte in the drop of blood.
• Transducer: Specifically, it converts the recognition of the bioreceptor into a
measurable signal. Most modern-day glucose meters and continuous glucose monitors
measure electrical signals, although earlier generations of glucose meters used a
colorimetric process (color change) that was measured optically.
• Electronics and display: These components process the transduced signal and prepare
it for display. The processed signals are then quantified and shown on either the glucose
meter’s display or the receiver for a continuous glucose monitor.

BLOOD GAS ANALYZERS

Blood Gas Analyzers

Blood gas analyzers are mainly used to measure the partial pressures of hydrogen (pH),
carbon dioxide (pCO2) and oxygen (pO2) present in the human blood. These measurements
are very useful to determine the acid base balance in the body. The electrodes used in the above
measurements are discussed in the chemical electrodes.

Table shows the normal values of pH, pCO2 and pO2 in human blood. A blood pH
below 7.35 indicates respiratory acidosis which indicates the respiratory failure. Meanwhile
the pCO2 of arterial blood is also increased to 90 mm of Hg. The respiratory failure can be
corrected temporarily using a ventilator.

Table -Normal blood gas parameters

Parameter Arterial blood Venous blood

PH 7.37-7.44 7.35-7.45

pCO2 Men 34-35 mm Hg 36-50 mm Hg

Women 31 - 42 mm Hg 34-50 mm Hg

pO2 75-90 mm Hg 25-40 mm Hg

 Similarly, when pH is raised to 7.60 and pCO2 is reduced to 18mm Hg, then there is
respiratory alkalosis which can be treated by setting of the ventilator so as to reduce the
ventilation. In the case of metabolic alkalosis where pH ≈ 7.5 and pCO2≈ 50 mmHg, the
infusion of NaCl and KCl gives better relief from it.

pH meter
According to the Goldman equation, the electrolyte membrane potentials are proportional to
the logarithm of the ion concentration and also to the absolute temperature of the electrolyte.
Thus, in a solution containing the hydrogen ion, a membrane separating two solutions has a
potential proportional to the hydrogen [H+] ion concentration

Thus, at a given temperature, 25°C,

Vm = -60 log [H+] + C (in mV)

where C is a constant

Since pH = - log [H*],

Vm = 60 pH + C

Usually, pH meters are calibrated so that the effect of the constant 'C' is cancelled. Thus, the
output of the meter directly gives the value of pH. One pH unit corresponds to 60 mV. Figure
shows the digital pH meter circuit. It is used to measure pH not only a given temperature but
also at different temperatures. The pH meter consists of a pH electrode which consists of a
glass (active) electro terminal and reference terminal. The calomel or silver-silver chloride
electrode in potassium chloride electrolyte is acting as a reference terminal. A salt bridge
consisting of a fiber wick saturated with KCI is at the tip of the reference electrode and keeps
the reference terminal potential essentially the same regardless of the solution under test. The
active terminal is sealed with common glass except for a tip made of pH sensitive glass which
consists of a hydrated gelatinous glass layer. Its membrane potential is proportional to the log
[H+] and therefore is proportional to the pH of the solution under test. Both the electrodes are
kept in a single glass enclosure.
The internal resistance of a glass electrode is very high (107 - 1010 ohms). Even though the
output from the pH electrode is high enough to deflect the voltmeter needle, to increase the
sensitivity and accuracy, the electronic circuitry is adopted. There is an external reference
voltage, to compensate the various errors and is also added with the output from pH electrode.
Further to determine the pH at different temperature a voltage from the temperature regulator
circuit corresponding to a given temperature is also added with the output from pH electrode.
The operational amplifiers amplify these voltages in the required manner and the final output
is given to a digital voltmeter. In the digital voltmeter, the display is obtained in terms of pH
as discrete numerals. Further the digital output may be used for further processing of signals.

Using pH electrode, pCO2 and PO2 can also be measured.

COLORIMETER
Colorimeter
• A colorimeter is a device used to test the concentration of a solution by measuring its
absorbance of a specific wavelength of light.
Principle of Colorimeter:
• When a beam of incident light of intensity I0 passes through a solution, following events
occur:
I0 = Ir + Ia + It
• A part of incident light is reflected. It is denoted by Ir
• A part of incident light is absorbed. It is denoted by Ia
• Remaining incident light is transmitted. It is denoted by It
Beer’s Law
This law states that the amount of light absorbed is directly proportional to the concentration
of the solute in the solution.
Lambert’s law
The Lambert’s law states that the amount of light absorbed is directly proportional to the length
and thickness of the solution under analysis.
Beer-Lambert’s law
The working principle of the colorimeter is based on Beer-Lambert’s law which states that the
amount of light absorbed by a color solution is directly proportional to the concentration of the
solution and the length of a light path through the solution.
A ∝ cl
Where,
A = Absorbance / Optical density of solution
c = Concentration of solution
l = Path length
A = ∈cl
∈ = Absorption coefficient

Light Source – The most common source of light used in colorimeter is a tungsten filament.
Monochromator – To select the particular wavelength filter or monochromators are used to
split the light from the light source.
Condenser lens - The light (tungsten lamp) passes through a condensing lens.
Filter - Filters helps to split a beam of light into different wave lengths allowing only the
required wavelength to pass through it reach the solution.
Sample holder – Test tube or Cuvettes are used to hold the color solutions.
Photo Detector System – when light falls on the detector system, an electric current is
generated, this reflects the Galvanometer reading.
Measuring device – The current from the detector is fed to the measuring device, the
Galvanometer, shows the meter reading that is directly proportional to the intensity of light.

Working of colorimeter
• Colorimeter measures absorbance and wavelength between 400 to 700 nm i.e. from the
visible spectrum of light of the electromagnetic spectrum.
• The light (tungsten lamp) passes through a condensing lens.
• And the filter helps to split a beam of light into different wave lengths allowing only
the required wavelength to pass through it reach the solution.
• When the beam of light reaches solution, it is transmitted, reflected, and absorbed by
the solution.
• The transmitted rays fall on the photo cell system where it measures the intensity of
transmitted light.
 • Photocell converts it into the electrical signals and send it to the galvanometer.
• The electric signals measured by the galvanometer are displayed in the digital form.

SODIUM POTASSIUM ANALYZER/ FLAME PHOTOMETER

Flame photometry is one of the branches of atomic absorption spectroscopy. It is also known
as flame emission spectroscopy. Flame photometer can be used to determine the concentration
of certain metal ions like sodium, potassium, lithium, calcium and cesium etc.

Principle of Flame photometer

The compounds of the alkali and alkaline earth metals dissociate into atoms when introduced
into the flame. Some of these atoms further get excited to even higher levels. But these atoms
are not stable at higher levels. Hence, these atoms emit radiations when returning back to the
ground state. These radiations generally lie in the visible region of the spectrum. Each of the
alkali and alkaline earth metals has a specific wavelength.

Parts of flame photometer

A simple flame photometer consists of the following basic components:
Source of flame: A Burner in the flame photometer is the source of flame. It can be maintained
in at a constant temperature. The temperature of the flame is one of the critical factors in flame
photometry.
a. Fuel gases: comprising the fuel reservoir, compressors, pressure regulators and pressure
gauges.
b. Atomizer: consisting, in turn, of a sprayer and the atomization chamber, where the aerosol
is produced and fed into the flame.
c. Burner: receives the mixture of the combustion gases.
d. Flame: the true source of emission

Nebulizer:
Nebulizer is used to send homogeneous solution into the flame at a balanced rate.

Optical system:
The optical system consists of convex mirror and convex lens. The convex mirror transmits the
light emitted from the atoms. Convex mirror also helps to focus the emissions to the lens. The
lens helps to focus the light on a point or slit.

Simple color filters:

The reflections from the mirror pass through the slit and reach the filters. Filters will isolate
the wavelength to be measured from that of irrelevant emissions.

Photo-detector:
The intensity of radiation emitted by the flame is measured by photo detector. Here the emitted
radiation is converted to an electrical signal with the help of photo detector. These electrical
signals are directly proportional to the intensity of light.
 • The solvent is first aspirated to obtain fine solid particles.
• These molecules in the solid particles are moved towards the flame to produce gaseous
atoms and ions.
• These ions absorb the energy from the flame get excited to high energy levels from the
ground state.
• But as these ions are unstable, they return back to ground state. While returning they
emit characteristic radiation.
• The intensity of emitted light is proportional to the concentration of the element.

SPECTROPHOTOMETER

Spectrophotometer
• The spectrophotometer refers to an instrument that measures the absorbance of the test
sample at a specific wavelength by measuring the amount of light transmitted by the
sample.
• This device contains several components like a light source, collimator,
monochromator, cuvette, light detector, and digital meter.
 • Spectrophotometry is a science which deals with the study of light intensity absorbed
or transmitted by the different solutions once a ray of light moves through the test
sample.
• The spectrophotometer principle depends upon the Beer-Lambert law.
Principle
Beer’s Law
This law states that the amount of light absorbed is directly proportional to the concentration
of the solute in the solution.
Lambert’s law
The Lambert’s law states that the amount of light absorbed is directly proportional to the length
and thickness of the solution under analysis.
Beer-Lambert’s law
The working principle of the spectrophotometer is based on Beer-Lambert’s law which states
that the amount of light absorbed by a color solution is directly proportional to the
concentration of the solution and the length of a light path through the solution.
A ∝ cl
Where,
A = Absorbance / Optical density of solution
c = Concentration of solution
l = Path length
A = ∈cl
∈ = Absorption coefficient

Spectrophotometric Instrumentation
This instrument consists of the following components:
Radiation source
It provides the continuous beam of EMR radiations in the required wavelength.
Monochromator
It is used to disperse the light into its components and then to choose a narrow range of
wavelengths for analysis.
Sample holder
It depends upon the physical appearance of the sample, Such as in UV vis the dilute solution
is taken in cuvettes
Detectors
It is a device, usually that converts the radiant energy into an electric signal.
Readout device
The electronic devices that will display the response of the detector

Working:
Single beam Spectrophotometer
• A beam of light from the light source falls onto the collimator convex lens and moves
to the diaphragm.
• The diaphragm ensures 100% transmittance and allows the light to fall onto the
monochromator device.
 • Monochromator device allows the transmittance of a single source of light onto the
focusing convex lens.
• The focusing convex lens transmits light of a particular wavelength from the sample to
the photocell detector.
• A photocell detects the portion of light transmitted or absorbed and gives the reading
on the display meter.

Double beam spectrophotometer

• In this, a fraction of light coming from the monochromator device parts into two beams.
One falls onto the reference sample and the other onto the test sample. Its mechanism
is more or less similar to a single beam spectrophotometer but differs because the dual
mirrors divide a single beam of light into two.
• One beam of light passes from the test sample to the photocell, and the other passes
from the reference sample to another photocell. A photocell detects the amount of light
transmitted or absorbed and gives the reading on the display meter.

BLOOD CELL COUNTER

Blood cell counter /Hematology analyzers are used to count and identify blood cells at high
speed with accuracy.

3 Part Diff. blood cell counter:

 • Neutrophils
• Lymphocytes
• Mid Cells (counting Monocytes, Basophils & Eosinophils)

Basic Operating Principles of 3 Part Diff. blood cell counter:

• Electrical Impedance Method
• Reflectance Photometry

5 Part Diff. blood cell counter:

• Lymphocytes
• Monocytes
• Polynuclear Basophils (PB)
• Polynuclear Neutrophils (PN)
• Polynuclear Eosinophils

Basic Operating Principles of 5 Part Diff. blood cell counter:

• Electrical Impedance Method
• Reflectance Photometry
• Flow cytometry

Electrical impedance
• Cell counting & sizing is based on the Coulter principle - detection & measurement of
changes in electrical impedance (resistance) produced by a blood cell as it passes
through an electrical field.
• When the cell passes through the aperture, flow of current is impeded and a voltage
pulse is generated. The no of pulses indicates the number of the blood cells.
• The amplitude (height) of each pulse is proportional to the cell volume.

Reflectance photometry
1. Light Source: A light source is directed through the flow cell
 2. Flow Cell: The prepared blood sample is introduced into a flow cell, which is a small
chamber or cuvette through which the sample flows in a controlled manner.
3. Photodetectors: Photodetectors are used to detect the light that is either scattered or
transmitted through the blood cells.
4. Reflectance Measurement: When the detectors measure the amount of light that is
scattered or reflected back from the blood cells. The amount of light reflected is
influenced by the properties of the blood cells, such as their size, shape, and optical
density.

Flow cytometry
Optical flow cytometry sensing

• The optical cytometry sensor consists of a quartz sensing sheath designed with a
hydrodynamic focusing region and cell path region that passes only a single cell at time.

Focusing is done by decreasing the diameter of the aperture.

• Light source is (He-Ne) Laser
• Two Photodetectors (photosensors)
– Photodetector A detects forward scatted light
– Photodetector B detects orthogonal scatted light
• blood sample enters the analyzer
– Optical counter → WBC count
– Colorimeter → hemoglobin
– Optical flow sensor → RBC count
AUTO ANALYZER (SIMPLIFIED SCHEMATIC DESCRIPTION)
• An autoanalyzer is medical lab equipment that conducts tests on blood, plasma,
cerebrospinal fluid, or urine and helps to study and evaluate the characteristics and
chemical components of the test sample.
• It is widely used in the medical industry for diagnosis and research purposes.
• The analyzer is based on the Lambert-beer law. This law states that there is a linear
relationship between the concentration and the absorbance of the solution, with the help
of which, the concentration of a solution to be calculated is measured.

Continuous flow type (pipeline type)

It means that the chemical reaction of each sample to be tested and reagents mixed in the same
pipeline is completed after the determination of the same item. This is the first generation of
automatic biochemical analyzer.
Advantage:
• It is possible to test large number of specimens for a particular test, accurately and
precisely, in a short duration.
Disadvantage:
• At a time, only one type of determination is performed by single channel continuous
flow analyzer.
• This autoanalyzer occupies a larger space in the laboratory.

Discrete
It means that the chemical reactions of each sample to be measured mixed with reagents
are completed in their respective reaction cups
Semi-automatic biochemical analyzer
In the analysis process (such as sample addition, holding, inhalation colorimetric,
results recording and other steps) part of the operation needs to be done manually, while other
operations can be done automatically by the instrument, this type of instrument is called semi-
automatic biochemical analyzer.
Semi-automated:
• Pipetting of reagent
• Pipetting of specimen
• Mixing and incubating the reaction mixture
Fully automatic biochemical analyzer
The entire process from sample addition to results is completed automatically by the
instrument, the operator only needs to put the sample on the specific position of the analyzer
and select the program to start the instrument to take the test report without manual intervention
during the period, this type of instrument is called fully automatic biochemical analyzer.
• Automatic dispensing of reagents
• Automatic dispensing of samples
• Automatic mixing of reaction mixtures
• Incubating of reaction mixtures, etc

Components of autoanalyzer
The automated system consists of a group of modular instruments interconnected together by
a manifold system and electrical systems. The various sub-systems are
• Sampling unit
• Proportioning pump
• Manifold • Dialyzer
• Heating bath or constant temperature bath
• Colorimeter/flame photometer/Fluorometer
• Recorder • Function monito

Sampler
• aspirates sample, standards, and wash solutions to the autoanalyzer system.
Proportioning pump and manifold
• introduces (mixes) samples with reagents to affect the proper chemical reaction to be
read by the colorimeter.
• pumps fluids at precise flow rates to other modules, as the proper color development
depends on reaction time and temperature.
Dialyzer
• separates interfacing substances from the sample material by permitting selective
passage of sample components through a semi permeable membrane.
Heating bath
• heats fluids continuously to exact temperature (typically 37°C incubation equivalent
temperature). Temperature is critical to color development.

Colorimeter
• monitors the changes in optical density of the fluid stream flowing through tubular flow
cell.
• Color intensities (optical densities) proportional to substance concentrations, are
converted to equivalent electrical voltages.
Recorder
• converts optical density electrical signal from the colorimeter into a graphic display on
a moving chart.

BIO SENSORS - PRINCIPLES - AMPEROMETRIC AND VOLTOMETRIC

TECHNIQUES.

Biosensor
• A Biosensor is an analytical device that detects changes in biological processes and
converts them into an electrical signal.
• The term biological process can be any biological element or material like enzymes,
tissues, microorganisms, cells, acids, etc.
• So, a Biosensor is a combination of a biological sensing element and a transducer,
which converts the data into electrical signals.
• Additionally, there will be an electronic circuit which consists of a Signal Conditioning
Unit, a Processor or Microcontroller and a Display Unit.
Components of biosensors
• The combination of biological sensitive element and a transducer will convert the
biological material into a corresponding electrical signal.
• Depending on the type of enzyme, the output of the transducer will be either current or
voltage. If the output is voltage, then well and good.
• But if the output is current, then this current should be converted into equivalent voltage
(using an Op-Amp based current to voltage converter) before proceeding further.
• The output voltage signal is usually very low in amplitude and superimposed on a high
frequency noise signal. So, the signal is amplified (using an Op-Amp based Amplifier)
and then passed through a Low Pass RC Filter.

• This process of amplifying and filtering the signal is the job of a Signal Processing
Unit or a Signal Conditioning Unit.
• The output of the signal processing unit is an analog signal that is equivalent to the
biological quantity being measured.
• The analog signal can be displayed directly on an LCD display but usually, this
analog signal is passed to a Microcontroller, where the analog signal is converted into
digital signal, since it is easy to analyze, process or store a digital signal.

Principle of biosensor
• The desired biological material is usually in the form of an enzyme.
• By a process known as Electroenzymatic approach, which is a chemical process of
converting the enzymes into corresponding electrical signals (usually current) with the
help of a transducer.
• One of the commonly used biological response is the oxidation of the enzyme.
Oxidation acts as a catalyst and alters the pH of the biological material.
• The change in pH will directly affect the current carrying capacity of the enzyme, which
is once again, in direct relation to the enzyme being measured.
• Output of the transducer i.e., the current, is a direct representation of the enzyme being
measured. The current is generally converted into voltage so that it can be properly
analyzed.
Types of biosensors

Electrochemical Biosensor

• The working of electrochemical biosensors is mainly based on the use of a biological

component/bio-receptor element which is in direct contact with an active transducer
(electrode) to obtain an analytical useful signal by coupling biochemical and
electrochemical interactions.
• The principle of electrochemical sensors is that when an electroactive analyte is
exposed to a fixed or varying potential of some predefined patterns which causes
oxidation or reduction of analyte on the working electrode surface, this leads to the
generation of electrochemical signal which can be measured by fluctuation on electron
flux. This signal can be detected electrochemically.

Amperometric Biosensor
 • These biosensors are based on the movement of electrons as a result of enzyme-
catalyzed redox reactions.
• The substrate or product can transfer electrons. This results in change in current flow
that can be measured.
• The magnitude of current is proportional to the substrate concentration.

Voltometric Biosensor

• This communication is the base of a new voltometric biosensor to notice acrylamide.

• This biosensor was built with a carbon glue electrode customized with Hb
(hemoglobin), which includes four prostatic groups of the hem (Fe).
• This type of electrode shows a reversible oxidation or reduction procedure of Hb (Fe).