ASIAN

COLLEGE OF ENGINEERING AND TECHNOLOGY

COIMBATORE – 641 110

ANNA UNIVERSITY - CHENNAI

REGULATION 2021

II YEAR / IV SEMESTER

DEPARTMENT OF BIOMEDICAL ENGINEERING

BM3411-BIOMEDICAL INSTRUMENTATION LABORATORY

Prepared by

Mr. R. Bharani Dharan, M.E.,

Assistant Professor/BME

Page1
 BM3411 BIOMEDICAL INSTRUMENTATION LABORATORY LTPC

0031.5

CYCLEI

1. Design of pre amplifiers to acquire bio signals along with

impedance matching circuit using suitable IC‘s.
2. DesignofECGAmplifierswithappropriatefiltertoremovepowerlineandot
her artifacts.
3. Design of EMG amplifier
4. Design a suitable circuit to detect QRS complex and measure heart rate.
5. Design off rontal EEG amplifier.
6. Design of EOG amplifier to detect eye blink.
7. Design a right leg driven ECG amplifier.

CYCLE II

8. Design and study the characteristics of optical Isolation amplifier.

9. Design a Multiplexer and Demultiplexer for any two bio signals.
10. Measurement of pulse-rate using Photo transducer.
11. Measurement of pH and conductivity.
12. Measurement of blood pressure using sphygmomanometer.
13. Measurement and recording of peripheral blood flow.
14. Design a PCB layout for any bio amplifier using suitable software tool.

TOTAL:60 PERIODS

Page2
 LIST OF EXPERIMENTS

S. No Name of the Experiment Date Pag Marks Sign

e
No
1. Design of pre amplifiers to acquire bio
Signals along with impedance
matching
2. Design of ECG Amplifiers with
appropriate
Filter to remove power line and other
3. Design of EMG amplifier

4. Design a suitable circuit to detect QRS

Complex and measure heart rate.
5. Design off rontal EEG amplifier.

6. Design of EOG amplifier to detect eye

blink.
7. Design a right leg driven ECG
amplifier.
8. Design and study the characteristics of
optical Isolation amplifier.

9. Design a Multiplexer and

Demultiplexer for any two bio signals.

10. Measurement of pulse-rate using

Photo
Transducer.
11. Measurement of pH and conductivity.

12. Measurement of blood pressure using

Sphygmomanometer.
13. Measurement and recording of
peripheral
Blood flow.
14. Design a PCB layout for any bio
amplifier
Using suitable software tool.

Page3
Page4
 Circuit Diagram:

Tabulation:

Output
Output voltage
Input Voltagein voltage with
S. No without output
(v) output
impedanceout(v)
impedanceout
(v)

Page5
Expt.No:01
DESIGN OF PRE AMPLIFIERS TO ACQUIRE BIO SIGNALS ALONG WITH IMPEDANCE
MATCHING CIRCUIT USING SUITABLE IC‘S
Date:

Aim:
To design pre amplifiers to acquire bio signals using suitable ICs with an
impedance matching circuit.

Apparatus required:

S. No Component Specification Quantity

1 Bio potential - 2
electrodes
2 IC741 - 2
3 resistors 10KΩ,10MΩ, 2,2,2
1MΩ
4 Capacitors 0.1µF,0.47µF, 1,1,1
1µF
5 RPS (0-30) V 1
6 Function (0-3) MHz 1
generator
6 DSO (0-8) MHz 1
7 Breadboard - 1
8 Connecting - As required
wires

Theory:
Potential differences originating from electrophysiological activity in the body may be
detected with this circuit by attaching bio electrodes and measuring the potential
differences between them. If the sensing bio electrodes are placed in the proximity of a
bio potential source, electrical interference induced from the power line or originating
from other sources of bio potentials in more remote parts of the body will affect both
probes more or less equally. The changes of signal detected simultaneously by both
electrodes are rejected by the first stage of the preamplifier. This stage is made up by
op-amp IC1 wired as a differential amplifier.
For low-frequency signals, the gain of the differential stage is given by
𝐺= 𝑅 𝑅4 10𝑀
𝑅
3 = =1000
Ω
𝑅1
=
10𝐾
2 Ω
At high frequencies, however, C1 has low impedance, forcing the first stage to act as a
low pass filter. In addition to limiting the bandwidth of the amplifier, C1 and C3 damp
oscillations and in stabilities of the circuit. Note that a gain of 1000 requires that large
dc offset voltages not be present on the bio potential signal. At this gain, the circuit will
stop operating if the offset voltage exceeds a mere 10mV. If higher offset voltages are
expected, the gain of the amplifier formed around IC1 must be decreased. For example,
to use this circuit as part of a surface ECG amplifier, the gain must be
recalculatedtocopewithoffsetpotentialsofupto±300mV.TheoutputofIC1isac-
coupledviaC2 to IC2. The -3-dB cutoff for the high-pass filter formed by C2 and R6 is
approximately 0.16 Hz. The filtered signal is then buffered by unity-gain voltage
follower IC2. To minimize electrical interference, the circuit should be built with a
compact layout on an appropriate printed circuit board or small piece of strip board.
The construction of the circuit is straight forward, but care must be taken to keep
wiring as short and clean as possible. Leads to the electrodes are coaxial cables with

Page6
their shields connected to ground at the circuit board.

Procedure:
First, test and calibrate the circuit. You will need a two-channel oscilloscope and a
signal generator. Take the following steps:

Page7
1. Connect the oscilloscope and the signal generator to the bio potential amplifier as in
non-inverting mode.
2. Applya10-Hzsignalof1-mVamplitudeasmeasuredbychannel2oftheoscilloscope.
3. Verify that the output signal is an amplified version of the input signal.
4. Determine the theoretical gain of the equivalent circuit and confirm that the output
signal has amplitude of Gnoninverting_1mV.
5. Without changing the settings of the instruments, connect the oscilloscope
and the signal generator to the bio potential amplifier as in inverting mode.
6. Verify that the output signal is an amplified and inverted (opposite phase) version of
the input signal. Determine the theoretical gain of the equivalent circuit and confirm
that the output signal has amplitude of Ginverting_1mV.

To measure the CMR, do the following:

1. Connect the equipment as shown in Figure.
2. Adjustthesignalgeneratortoproducea60-Hz5-Vp-pcommon-modeinputsignalVinCM.
3. Measure the corresponding common- mode output voltage VoutCM.

Result:
Thus, the design of pre-amplifiers to acquire bio signals using suitable ICs with
an impedance matching circuit was done.

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Page9
Circuit diagram:

Tabulation:

Output voltage Output voltage

S. No Input Voltagein(v)
without filter, with filter,
Vout(v) Vout(v)

Page10
Expt.No:02
DESIGN OF ECG AMPLIFIERS WITH APPROPRIATE FILTERTORE MOVE POWER
LINE AND OTHER ARTIFACTS
Date:

Aim:
To design an ECG amplifier with appropriate filter to remove power line and other
artifacts.

Apparatus required:

S. No Component Specification Quantity

1 ECG - 1
electrodes
2 AD620A - 1
3 OP97 - 1
4 resistors 220kΩ,1MΩ, 3,1,1,2,1
10kΩ,22kΩ,
6.98kΩ
5 Capacitors 0.1µF 1
6 RPS (0-30) V 1
7 Function (0-3) MHz 1
generator
8 DSO (0-8) MHz 1
9 Bread/PCB - 1
Board
10 Connecting - As required
wires

Theory:
The depolarizing field in the heart is a vector which alters its direction
and magnitude through the cardiac cycle. The placement of the electrodes
on the surface of a patient determines the view which will be obtained of
that vector as a function of time. The most commonly used electrode
placement scheme is shown in Figure 1. Here the differential potential is
measured between the right and left arm, between the right arm and the
left leg and between left arm and left leg. These three measurements are
referred to as leads I, II, III respectively. This measurement lead
placementwasdevelopedbyEinthovenwhostatedthatthroughmeasuremento
f leadIandleadIIthesignalseenatleadIIIcouldbecalculated.Thisisthemost
basic form of ECG lead placement: from this the various features of the
heart’s depolarization can be calculated. Clinically there is a range of lead
placement schemes which incorporate limb leads and chest leads.

Page11
Figure1

Page12
Therefore, the ECG waveform shows the clinician the electrical
waveforms associated with the contraction of the atria and ventricles.
From an ECG a clinician may determine the relative timing of the
contractions of the atria and the ventricles and assess the relative
amplitude of the atrial and ventricular depolarization and depolarization.
This information may allow the identification of mild heart block.
Following a heart attack, a patient’s ECG shows changes as the timing
and shape of the waveform are dependent on the transmission of the
waveform through the muscle tissue. These changes with ischemic muscle
damage associated with heart attacks.

The amplifiers we use in biomedical engineering, data acquisition or

where the signal of interest is represented by a small voltage fluctuation
superimposed on a voltage offset are called Instrumentation amplifiers.
Instrumentation amplifiers have a high CMRR (Common Mode Rejection
Ratio) which means they have the ability of a differential amplifier to not
pass (reject) the portion of the signal common to both the
+and–inputs.

The noise comes from muscle contractions, power line interference 50-60
Hz, electrode contact noise, noise from other electronic devices and etc.
The filter for the ECG application should be a notch filter (high-pass and
low-pass filter). It should filter in the range from 0.5 Hz to 50 Hz. In the
given circuit simple RC high pass and low pass filter, in series are
connected (just two capacitors and resistors).

Procedure:
1. Connect the circuit as per given diagram.
2. Measure the input voltage signal.
3. Measure the output signal using multimeter and tabulate to check
the correspondence between original input and filtered output.

Result:
Thus, an ECG amplifier with appropriate filter to remove power line and other
Page13
artifacts was designed and the appropriate values are measured.

Page14
Circuit Diagram:

Sliding theory:

Block diagram:

Page15
Ex. No:03
DESIGN OF EMG AMPLIFIER
Date:

AIM:
To record the EMG signals in resting and contracted state by designing an EMG
amplifier.

EQUIPMENTANDACCESSORIESREQUIRED:

S. No Name of the Component Specification Quantity

1. Bio signal simulator - 1
2. IN128–Instrumentation amplifier - 1
3. SMD/SMT–Schott key diode - 1
4. Capacitors 47µF 1
5. RPS (0-30) V 1
6. Digital Storage Oscilloscope (0-8) MHz 1
- As
7. Connecting Cables
required
8. Bread/PCB board - 1

THEORY:
Generation of EMG
The EMG is generated when a motor neuron action potential from the spinal
cord arrives at a motor end plate. Its arrival causes a release of ACh
(Acetylcholine) at the synaptic cleft (1) which causes a depolarization (Action
Potential). This action potential electrically travels downward from the surface in
a transverse tubule (2). This in turn causes a release of Ca++ (3), causing cross-
bridge binding (4) and the sarcomere of the muscle to contract (5).
An Electromyography (EMG) is a measurement of the electrical activity in
muscles as a byproduct of contraction. An EMG is the summation of action
potentials from the muscle fibers under the electrodes placed on the skin. The
more muscles that fire, the greater the amount
ofactionpotentialsrecordedandthegreatertheEMGreading.Thetestisusedtohelp
detect neuromuscular abnormalities. During the test, one or more small needles
electrodes are inserted through the skin into the muscle.
The contraction of the skeletal muscle results in the generation of action
potentials in the individual muscle fibers, a record of which is known as
electromyogram. In this case skeletal muscle re polarization takes places more
rapidly, the action potential lasting only a few milliseconds. Since most of EMG
measurements are made to obtain an indication of the amount of activity of a
given muscle, or a group of muscles rather than an individual muscle fiber, the
EMG pattern is usually a summation of the individual action potentials from the
fibers constituting the muscles or muscles being studied. In voluntary
contraction of the skeletal muscle, the muscle potentials range from
50µVto5mVandthedurationfrom 2 to 15ms. The values vary with the Anatomical
position of the muscle and the size and location of electrode.

The super simple EMG muscle sensor described here will help make it easier for people play with
Page16
EMG. It uses a ultra-simple 3 piece EMG circuit, consisting of an INA128 Instrumentation Amplifier
(IA), a capacitor, and a rectifying diode to perform half wave signal rectification. The instrumentation
amplifier, amplifies differential inputs and subtracts the common mode signal (basically takes two

Page17
signals and subtracts what is common in both), which causes noise presented on both channels to be IA
gain pins, which sets a variable frequency dependent gain, thereby only amplifying higher frequencies,
and thus creating a similar affect to a high pass filter. The capacitors impedance (which is inversely
proportional to the IA amplification) is calculated from below equation

Impedance= 1/ (2∗ PI ∗Frequency∗ Capacitance)

The EMG sensor requires 3 electrodes, a negative, and positive and ground, with the positive and
negative placed adjacent to each other, and separated by a 1cm gap. These electrodes are placed over
the muscle you want to monitor, and the ground electrode placed over a bony area with minimal to no
muscle activation. The exact locations of the electrodes can be determined by trial and error. The
electrodes used were 30mm diameter, reusable, self-adhesive, and long-term electrodes often found in
TENS, EMG and Neuromuscular stimulation applications. The only other additions you will need to see
the EMG signal are a 5V positive and negative voltage source to power your sensor, and either an
oscilloscope, or an analogue pin from a favorite microcontroller partially suppressed. The super simple
EMG circuit uses a capacitors reactive impedance attached to the

PROCEDURE:
1. Connect the EMG signals to instrumentation amplifier
through medical simulator.
2. Record the EMG signals in resting state and contracted state through
DSO.

NORMALEMGSIGNAL:

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Page19
TABULATION:

NAMEOF CONTRACTIONSTATE
RESTINGSTATE
S.NO THE AMPLITUDE
AMPLITUDE WAVEFORM
SUBJECT (V)
(V)

RESULT:
Thus, the EMG waveform in resting and the contracted state were picked up
and analyzed using EMG recorder

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Page21
Normal ECG Wave

Conducting System:

Tabulation (ECG waveform):

Amplitude: Time period:

P wave–0.25mV PR interval–0.12–0.22sec
Q wave–0.4mV QT interval–0.35–0.44sec
R wave–1.6mV ST segment–0.05–0.15sec
T wave–0.1–0.5mV P wave interval–0.11sec
U wave-<0.1mV QRS duration–0.06–0.1sec

Page22
Expt.No:4
DESIGN A SUITABLE CIRCUIT TO DETECT QRS COMPLEX MEASURE
HEART RATE
Date:

AIM: To record and analyze the ECG for a subject in bipolar configuration and to
calculate the heart rate using different methods from the recorded chart paper.

EQUIPMENTANDACCESSORIESREQUIRED:

S. Name of the Equipment Quantity

No
1. ECG machine 1
2. Lead wires & electrodes 3-12max
3. ECG paper roll 1
4. Conductive gel 1

THEORY:
Electrocardiograph (ECG) is an instrument used to record electrical
activity of the heart over a period of time, as detected by electrodes attached to
the surface of the skin and recorded by a device external to the body. The
recording produced by this noninvasive procedure is termed an
electrocardiogram.
An ECG is used to measure the rate and regularity of heart beats, as well
as the size and position of the chambers, the presence of any damage to the
heart, and the effects of drugs or devices used to regulate the heart, such as a
pacemaker. The conducting system of the electrical pulses in heart is shown
above. ECG provides a wide range of cardiac disorders such as the presence of
an inactive part (infarct) or an enlargement (hypertrophy) of heart muscle. The
ECG device detects and amplifies the tiny electrical changes on the skin that are
caused when the heart muscle depolarizes during each heartbeat.
The diagnostically useful frequency range is usually 0.05 too 150Hz. The
interference of non-biological noises can be handled by using modern differential
amplifiers, which are capable of providing excellent rejection capabilities.
Common Mode Rejection Ratio of the order of 100-120db with 5KΩ unbalance in
the leads is a desirable feature of ECG machines. It is necessary to use a notch
filter tuned to 50Hz to reject hum due to power mains.

LEADS:
The term “lead" in electro cardiograph refers to three electrical cables attaching
the electrodes to the ECG recorder. These leads are placed according to a
practical system of electrocardiography used in medical diagnostics known as
Einthoven’s Triangle. Einthoven’s Triangle is an equilateral triangle whose
vertices lie at the left and right shoulders and the pubic region and whose center
corresponds to the vector sum of all electric activity occurring in the heart at
any given moment, allowing for the determination of the electrical axis.
Einthoven's triangle is approximated by the triangle formed by the axes of the
bipolar electrocardiographic (ECG) limb leads I, II, and III.

Page23
There are three different configurations of leads that are used to record ECG
signals. They are:
1. Limb leads
2. Pre cordial leads
3. Augmented limb leads

Page24
Fig: Einthoven Triangle

Fig: Lead system

Fig: Lead system

Page25
LIMBLEADS:
Leads I, II and III are called limb leads. The electrodes that form these
signals are located on the limbs—one on each arm and one on the left leg. The
limb leads form the points of what is known as Einthoven's triangle.
Lead I is the voltage between the (positive) left arm (LA) electrode and right arm
(RA) electrode:

Lead II is the voltage between the (positive) left leg (LL) electrode and the right
arm(RA) electrode:

Lead III is the voltage between the (positive) left leg (LL) electrode and the left arm
(LA) electrode:

AUGMENTEDLIMBLEADS:

Leads aVR, aVL, and aVF are augmented limb leads. They are derived from
the same three electrodes as leads I, II, and III. However, they view the heart
from different angles (or vectors) because the negative electrode for these leads
is a modification of Einthoven's central terminal. Wilson Einthoven's central
terminal paved the way for the development of the augmented limb leads aVR,
aVL, aVF and the pre cordial leads V1, V2, V3, V4, V5 and V6.
 Lead augmented vector right (aVR) has the positive electrode (white)
on the right arm. The negative electrode is a combination of the left arm
(black) electrode and the left leg (red) electrode, which "augments" the
signal strength of the positive electrode on the right arm.
 Lead augmented vector left (aVL) has the positive (black) electrode on
the left arm. The negative electrode is a combination of the right arm
(white) electrode and the left leg (red) electrode, which "augments" the
signal strength of the positive electrode on the left arm.
 Lead augmented vector foot (aVF) has the positive (red) electrode on the left
leg. The negative electrode is a combination of the right arm (white)
electrode and the left arm (black) electrode, which "augments" the signal
of the positive electrode on the left leg.

PRECORDIALLEADS:

The electrodes for the pre cordial leads (V 1, V2, V3, V4, V5 and V6) are placed
directly on the chest. Because of their close proximity to the heart, they do not
require augmentation. Wilson's central terminal is used for the negative
electrode, and these leads are considered to be Unipolar. The pre cordial leads
view the heart's electrical activity in the so-called horizontal plane. The heart's
electrical axis in the horizontal plane is referred to as the Z axis.

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Page27
Fig: Lead Placement

PLACEMENT OF ELECTRODES:

Electrode
label (in the Electrode placement
USA)

RA On the right arm, avoiding thick muscle.

LA In the same location where RA was placed, button the left arm.

RL On the right leg, lateral calf muscle.

LL In the same location where RL was placed, button the left leg.

In the fourth intercostal space (betweenribs4and5) just to the right of

V1
the sternum (breastbone).
In the fourth intercostals space (between ribs 4 and 5) just to the
V2
left of the sternum.
V3 Between leads V2 and V4.

V4 In the fifth inter costal space (between ribs5 and 6) in the mid-
clavicular line.
V5 Horizontally even with V4, in the left anterior axillary line.

V6 Horizontally even with V4and V5in the mid auxiliary line.

Page28
EFFECT OF ARTIFACTS ON ECG RECORDING:
Abnormal patterns of ECG recording may be due to pathological states or on
occasion they may be due to artifacts.
 INTERFERENCE FROM POWRER LINE:
Power line interference is easily recognizable since the interfering voltage in
ECG would have a frequency of 50Hz. This interference may be due to the stray
interference of the alternating current on the patient or because of alternating
current fields due to loops in the patient cable. Other causes of interference are
loose contacts on the patient cable as well as dirty electrodes. When the
machines or the patient is not properly grounded, power line may even
completely obscure the ECG waveform. The most common cause of 50Hz
interference is the disconnected electrode resulting in a very strong disturbing
signal. It is often strong enough to damage the stylus of an unprotected direct
writing recorder, and therefore needs a quick action.

 SHIFTING OF BASELINE:
A wandering baseline but otherwise normal ECG trace is usually due to the
movement of the patient or electrode, the baseline shift is eliminated by
ensuring that the patient lies relaxed and the electrodes are properly attached.
Baseline wander is usually observed immediately after the application of
electrodes.

 MUSCLE TREMOR:
Irregular trembling of ECG trace, without wandering of the baseline occurs
when the patient is not relaxed or is cold. It is generally formed in the case of
older patients. Muscle tremor signals are especially bothersome on limb leads.
When a patient moves or the muscles are stretched. For normal ECG recording,
the patient must be advised to get warm and to relax so that muscle tremor from
shivering or tension is dominated. The most critical component of the ECG
recorder is the patient cable. They should be made of silicon-rubber, which
provides better elasticity over long period.

PROCEDURE
1. The patient has to remove any jewelers or other objects that may interfere
with the procedure.
2. The patient has to lie flat on a wooden table or bed in a supine position for
the procedure. It will be important for to lie still and not talk during the
procedure, so as to avoid artifacts.
3. If chest, arms or legs are very hairy, the technician may shave or clip small
patches of hair, as needed, so that the electrodes will stick closely to the
skin.
4. Electrodes will be attached to chest, arms, and legs after placing gel.
5. The lead cable will be attached to the surface electrodes.
6. The ECG will best start. It will take only a short time for the tracing to be
completed.

Page29
7. Once the tracing is completed, the technician will disconnect the leads and
remove the skin electrodes.
8. Selectthespeedat25mm/secandthecalibrationsignalat10mm/mv from ECG
recorder.
9. By changing the leads elector, draw the wave forms for LEAD I, LEAD II,
LEADIII, aVR,aVL, Avf and chest leads.

Page30
ECG WAVEFORM:

Page31
CALCULATION OF HEART BEAT:
Count the distance between successive ‘R’ waves. By using the formula, the heart
rate can be calculated
1500
Heart Rate (H.R) = ---------- beats/min
RR Interval
METHOD 1: When the rhythm is regular, the heart rate is 300 divided by the
number of large squares between the QRS complexes.
o For example, if there are 4 large squares between regular QRS
complexes, the heart rate is 75bpm (300/4=75).
300*0.2(timeof1largebox) =60sec.
METHOD2: The second method can be used with an irregular rhythm to
estimate the rate. Count the number of R waves in a 6 second strip and multiply
by 10.
o For example, if there are 7 R waves in a 6 second strip, the heart
rate is 70bpm (7x10=70).
METHOD 3: When the rhythm is regular, the heart rate is 1500 divided by the
number of small squares between the QRS complexes.
o For example, if there are 22 small squares between regular QRS
complexes, the heart rate is 68bpm (1500/22=68).
1 small square – 0.04 sec. For 60 sec the numbers of small boxes are 1500.
(1500*0.04 = 60sec)
METHOD4:
60 divided by [(number of small squares between the QRS complexes) *(0.04sec)]

RESULT:
Thus, the QRS complex is detected and heart rate is measured using ECG circuit.

Page32
Page33
Frontal EEG amplifier:

Waveform:

Page34
Ex. No:05 DESIGN OF FRONTAL EEG AMPLIFIER
Date:

AIM:
To design an EEG amplifier circuit with electrodes placed in frontal lobe and
observe the response.

APPARATUS REQUIRED:
S. No Name of the Component Specification Quantity
1. Medical simulator - 1
2. AD620–Instrumentation amplifier - 1
3. OP 90 / LM324 – - 2/1
Operational
Amplifier
4. Resistors 10MΩ,1kΩ, 2,2,1,1,2
390kΩ,10kΩ,
200kΩ
5. Capacitors 0.01µF, 2,1,1
0.1µF,0.47µ
F
6. Dual power supply (0-30) V 1

7. DSO (0-8) MHz 1

8. Breadboard - 1

THEORY:
EMG is measured using similar techniques to that used for measuring EKG, EEG
or other electrophysiological signals. Electrodes are placed on the skin overlying
the muscle. Alternatively, wire or needle electrodes are used and these can be
placed directly in the muscle signals are small and need to be amplified by an
amplifier designed to measure physiological signals. These amplifiers include a
differential amplifier circuit, and frequently include some filtering and other
signal processing features. The frequency range is 0-10khz and the amplitude
ranges from 0.1 to 0.5mv.

EEG amplitude and frequency bands:

EEG Signals are mainly classified on the basis of frequency. Frequency
information is particularly significant since the basic frequency of the EEG varies
greatly with different behavioral states. To assist in the EEG analysis, the normal
frequency range of the EEG (0.5Hz to 30Hz) has been divided into five bands.
Delta(δ) 0.5 - 4Hz
Theta(φ) 4 - 8Hz
Alpha(α) 8 -12Hz
Beta(β) 12 - 22Hz
Gamma(γ) 22 - 30Hz

Page35
Page36
EEG signal voltage amplitudes range from about 1 to 100 µV peak to peak at low
frequencies (0.5Hz) at the cranial surface. At the surface of the cerebrum,
signals may be 10times larger. Also, brain stern signals measured at the cranial
surface are often no larger then
0.25µV peak to peak (100-3000Hz). Weak EEG signals require input pre amplifiers
(differential type) that have high gain and internal/external noise rejection.
Alpha activity: -This activity is less than 10µV peak to peak, occurring data
frequency between 8 to 13 Hz. These signals at the posterior brain in the waking
person with eyes closed. Opening the eyes and focusing attention greatly
reduces alpha waves. Subject to a sleep, the alpha wave disappears completely.
Beta activity: - This activity is less than 20 µV peak to peak in the frequency
range of 13 to 22 Hz. It is available over the entire brain but is most
predominant over the central region. Gamma activity: - This activity is less than
2 µV peak to peak in the range of 22 to 33 Hz. This is a low-amplitude& high
frequency wave’s resulting from attention or sensory stimulation. The activity: -
This activity is less than 100µV peak to peak in the range of 4 to 8Hz. These
occur mainly in the parietal and temporal region in children. This also occurs in
some adults during disappointment and frustration. Delta activity: - This activity
in the frequency range of 0.5 to 4 Hz. They occur in deep sleep.

PROCEDURE:
1. Connect the components as per circuit diagram.
2. Place the electrodes in frontal lobe and observe the response in DSO.
3. Plot the response.

RESULT:
Thus, the EEG amplifier is constructed and responses are observed.

Page37
Page38
Structure of human eye:

Circuit Diagram:

Model graph:

Page39
Expt.No:06
DESIGN OF EOG AMPLIFIER TO DETECT EYE BLINK
Date:
Aim:
To design an EOG amplifier and detect the response of eye blink.

Apparatus required:
S. Name of the Component Specification Quantity
No
1. OP07–Precision amplifier - 2
2. AD620–Instrumentation amplifier - 2
3. Resistors 560Ω,1MΩ, 1,1,4,1
1kΩ,4.75kΩ
4. Capacitors 1µF,3.3µF, 1,1,1
4.7µF
5. RPS (0-30) V 1
6. DSO (0-8) MHz 1
7. Surface electrodes - 4
8. Connecting wires - As req

Theory:
The eye is a place of a steady electric potential field that is quite unrelated to
light stimulation. This field can be detected even with the eyes closed or eyes in
total darkness. This steady electric potential can be viewed as a fixed dipole with
positive pole at the cornea and negative pole at the retina. The magnitude of this
cornea retinal potential is in the range 0.4-1.0 mV. This potential is not
generated by excitable tissue but it is due to the occurrence of higher metabolic
activity in the retina. For the invertebrates, the polarity of this potential
difference is of opposite to that found in vertebrates such as human beings. This
cornea retinal potential is roughly aligned with the optic axis and hence rotates
with the direction of gaze, which can be measured by surface electrodes placed
on the skin around the eyes. Rotation of the eye and the cornea retinal potential
form the basis for a signal measured at a pair of per orbital surface electrodes.
The signal is known as the Electrooculogram (EOG).

Generation of EOG:
Due to the higher metabolic rate at the retina compared to the cornea, the eye
maintains a voltage of +0.40 to +1.0 mill volts with respect to the retina. This
cornea retinal potential, which is roughly aligned with the optic axis and (as a
result) rotates with the direction of gaze, can be measured by surface electrodes
placed on the skin around the eyes. The actual recorded potentials are small
either range of 15 to 200 micro volts and require amplification before
processing. With proper calibration, the orientation of the electric dipole can be
used to specify the angular position of the eyeball with optimum accuracy which
lies within 2 degrees vertically and 1.5 degrees horizontally.

Electrodes and input cables:

Disposable pre-gelled Ag/AgCl electrodes were used to acquire EOG signals from
the body. Since the EOG signal amplitude range was in microvolts, they were

Page40
very much susceptible to various noise sources. To overcome the effects of RF
noise and electromagnetic interference, shielded wires were used to connect
Ag/AgCl electrodes and data acquisition circuits.

Page41
Amplifier Circuitry:
In general, the EOG signal amplitude and band width varies in the range of 50-
3500 µV. USB-4704 is a 12-bit analog-to-digital converter. Hence, care had to be
taken to design a bio potential which would amplify the signal in such a way that
the digitization of the EOG signals result in minimum quantization error. To
ascertain this, the gain of the bio potential amplifier should be adjusted so that
100 µV EOG signal is amplified above 1.4 mV [20]. Also, the characteristics of
AD620 suggest that as the gain of bio-potential amplifier is increased, there was
a subsequent increase in the common-mode rejection ratio (CMRR). Taking the
above facts into consideration, the bio potential amplifier was designed with a
gain of 1500. Circuit diagram 31 of vertical EOG signal acquisition system is
given in Figure 4.1. Same circuit can be used to acquire vertical EOG signals.

Figure: Electrooculography electrode placements.

EOG signals were measured by placing electrodes on the region surrounding the
eye. They were recorded from two separate regions: horizontal and vertical.
Horizontal electrodes were for detecting horizontal eye movements (left and
right eye movement) and vertical electrodes were for detecting vertical
movements (up and down cornea movements) [18]. Three to five electrodes were
used to acquire the EOG signals. Figure 3.2 shows the five-electrode placement
in Electrooculography. Electrodes x+ and x- were horizontal electrodes and y+
and y- were vertical electrodes. The fifth electrode placed on the forehead was
for reference [2, 19].AD 620AN was used as the preamplifier. The AD620was a
low cost, high accuracy 27 instrumentation amplifier. It was used as a pre
amplifier because of its high CMRR and high input impedance. The main source
of noise present in the signal was power line noise and baseline wandering. Base
line wandering was due to the presence of low frequency components in the
signal. These two noises were removed by using a band pass filter with cut-off
frequency 0.5 to35 Hz.

Procedure:
1. Place the surface electrodes around the eye as per figure.
2. Connect the circuit as per given.
3. View their spouse of eye during movement and draw the graph.

Page42
Result:
Thus, EOG amplifier was designed and eye movements are tacked and plotted.

Page43
Circuit Diagram:

ECG waveform:

Page44
Expt.No:07
DESIGN A RIGHT LEG DRIVEN ECG AMPLIFIER
Date:
Aim:
To design a right leg driven ECG amplifier circuit.

Apparatus Required:
S. No Name of the component Specification Quantity
1 AD620–Instrumentation amplifier - 1
2 LF411-JFETinputopamp - 1
3 Resistors 5.5kΩ,10kΩ,20kΩ, 2,1,2,1
1MΩ
4 Capacitors 0.1µF 1
5 ECG electrodes - 3
6 DSO (0-8) MHz 1
7 RPS (0-30) V 1

Theory:
A Driven Right Leg Circuit or DRL circuit 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). Unfortunately,
the patient's body can also act as an antenna which picks up electromagnetic interference, especially
50/60Hz noise from electrical powerlines. 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.

This stage provides a reference point on the patient that normally is at ground potential. It is
implemented in ECG measurement systems to counter common mode noise in the body. This circuit is
as shown in Fig. The two signals entering the differential amplifier are summed then inverted and
amplified in the right leg driver before being feed-backed to an electrode attached to the right leg. The
other electrodes also pick up this signal and hence the noise is cancelled. Without the driven right leg
circuit, a direct ground path is provided, which hence introduce risk of a ground fault hazard. With the
right leg driven circuit provided, the body’s displacement current flows not to ground but rather to the
Op Amp output circuit as shown below.

Procedure:
1. Connect the components as per circuit diagram.
2. Connect the right leg to output of Op-amp and check the output
waveform.

Result:
Thus, right leg driven ECG circuit is constructed and responses are plotted.

Page45
Pinout Diagram– MC2TE

Circuit Diagram:

Response Diagram:

Page46
Ex. No:8
DESIGN AND STUDY THE CHARACTERISTICS OF OPTICAL ISOLATION AMPLIFIER
Date:

AIM:
To study of characteristics of optical isolation amplifier.

APPARATUS REQUIRED:
S. Name of the Component Specification Quantity
No
1. MC2TE–Opti isolator IC - 1
2. Resistor 1kΩ 1
3. Connecting wires - As reqd

THEORY:
This amplifier enables galvanic isolation between input and output circuits in any
electronic system and is intended for applications requiring safe, accurate
measurement of DC. and low-frequency, low level signals in the presence of high
common-mode voltage. Very high common-mode voltage (> 1000 v), high
isolation resistance 1011ohm) and high CMRR (-120 dB) are the required
characteristic of such an amplifier. Optically coupled isolation amplifier is
important for this purpose.
It makes use of the isolation properties of LED-photodiode or LED-
phototransistor coupler together with the OP AMP. In this coupler, LED is used
at the input and it is optically coupled to a photodiode or phototransistor
enclosed in the same package at the output terminals. As the coupling is only
through optical means, excellent input-output isolation is obtained by main
training the power supplies for these devices separate. While phototransistor
isolators have limited bandwidth (< 100 KHZ), the photodiode isolators facilitate
bandwidth of MHZ. However, the overall bandwidth is determined by the OP
AMP characteristics. A major problem in the operation or such an amplifier is
the non1inear transfer characteristic of optical coupling which introduces signal
distortion. Further, high temperature sensitivity of the optical coupling
introduces large gain drift with temperature.
A compensating, non-linear temperature sensitive feedback is used in
the isolated voltage amplifier circuit. Here, the current supplied to the
transmitting LED is controlled by a feedback LED-phototransistor coupling. The
two LEDs are supplied the same level or current, i.e., the value which makes the
feedback phototransistor accept the signal current from vin and the bias current
from V1. This current is independent of the non-linearity and thermal variations
of the feedback LED-phototransistor coupling. Considering the two couplers to
be well-matched, for the same power supply voltage level in the two sections
(i.e., V1 = V2).

MC2TE–Opti coupler IC:

MCT2Eis a phototransistor Optocoupler, as the name “phototransistor” suggests it
has a transistor which is controlled based on light (photon). So this IC basically has
an IR (infrared) LED and a photo-transistor inside it. When the IR led is powered the

Page47
light from it falls on the transistor and it conducts. The arrangement and pin-outs of
the IR LED and the photo-transistor is shown below.

Page48
This IC is used to provide electrical isolation between two circuits, one part of the
circuit is connected to the IR LED and the other to Photo-transistor. The digital
signal given to the IR LED will be reflected on the transistor but there will be no
hard electrical connection between the two. This comes in very handy when you are
trying to isolate a noisy signal from your digital electronics, so if you are looking for
an IC to provide optical isolation in your circuit design then this IC might be the
right choice for you.
TheMCT2Ecan be used in two modes, the Phototransistor mode and the photodiode
mode. Out of which the Phototransistor mode is mostly used, so let us look at that
mode first.
In the Photo-transistor mode we will not be using the base pin (pin 6) of the
transistor; we just have to connect the anode pin of the IRLED (pin1) to the logic in
put which has to be isolated and the cathode (pin 2) of the IR led to the ground.
Then Pull high the collector pin of the transistor using a resistor (here I have used
1K) and connect the collector pin to the output of your desired logic circuit. The
Emitter (pin 4) is grounded.
Note: The ground line of the IR LED (pin 2) and the ground line of the transistor
(pin 4) will not be connected together. This is where the isolation occurs.
Now, when the Logic input is low the IRLED will not conduct and hence the transistor
will also be in off state. Hence the Logic output will remain high, this high voltage can be
set anywhere up-to30V (Collector-Emitter Voltage) here I have used +5V. There pull-up
resistor 1Kactsasa load resistor.
But when the Logic input is made high, this high voltage should be a minimum of
1.25V (Diode Forward voltage) the IR LED conducts and so the photo-transistor is
also turned on. This will short the collector and emitter and hence the Logic Output
voltage will become zero. This way the logic input will be reflected at the logic
output and still provides and isolation between the two. The complete working can
also be understood form the GIF file above.
Another important parameter to consider while using an Optocoupler, is the rise
time (tr) and fall time (tf). The output will not get high as soon as the input logic is
made low and vice versa. The below waveform shows the time taken for the output
to transit from one state to another. For MCT2E the rise time (TPDHL) and fall time
(TPDLH) is 5us.
CHARACTERISTICS OF OPTICAL ISOLATION AMPLIFIER:
The characteristics of optical isolation amplifier are as follows:
Very high common-mode voltage (> 1000 v), The input common voltage is defined as
the average voltage at the inverting and noninverting input pins. For op amp,
CMVR is the range of common mode signal for which the amplifier's
operation remains linear. If the voltage present on the"-" input equal V1,
and the voltage on the"+" input equal V2, then the common mode voltage
is VCM = (V1+V2)/2.

Page49
 Very high common-mode rejection ratio(120db), The common-mode
rejection ratio is the rejection by the device of unwanted input
signals common to both input leads, relative other wanted difference
signal. It is defined as the ratio of the differential voltage
amplification to the common- mode voltage

Page50
 amplification. Ideally this ratio would be in invite with common mode
voltages being totally rejected.
 High isolation resistance (1011ohm)
Procedure:
1. Connect the components as per circuit diagram.
2. Check the output by changing input logic as per model response
diagram and plot the output.

RESULT
Thus, the characteristics of optical isolation amplifier are studied.

Page51
Page52
Multiplexer with digitally adjustable amplifier gain:

Demultiplexer with digitally adjustable amplifier gain:

Truth Table:
4:1channel multiplexer:

1:4channel demultiplexer:

Page53
Expt.No:09
DESIGN A MULTIPLEXER AND DEMULTIPLEXER FOR ANY TWO BIO SIGNALS
Date:

Aim:
To design an analog multiplexer and demultiplexer for bio signals.

Apparatus Required:
S. No Name of the component Specification Quantity
1 Medical simulator - 1
2 Resistors 1KΩ,10KΩ,100Ω, 1each
200Ω,500Ω
3 IC741 - 1
4 Dual RPS (0-30) V 1
5 DSO (0-8) MHz 1
6 Bread/PCB board - 1
7 Connecting wires - As required
8 SPST switch - 4

Theory:
Multiplexers are switching circuits that just switch or route signals through
themselves, and being a combinational circuit, they are memoryless as there is
no signal feedback path. The multiplexer is a very useful electronic circuit that
has uses in many different applications such as signal routing, data
communications and data bus control applications.
When used with a demultiplexer, parallel data can be transmitted in serial form
via a single data link such as a fiber-optic cable or telephone line and converted
back into parallel data once again. The advantage is that only one serial data line
is required instead of multiple parallel data lines. Therefore, multiplexers are
sometimes referred to as “data selectors”, as they select the data to the line.
Multiplexers can also be used to switch either analogue, digital or video signals,
with the switching current in analogue power circuits limited to below 10mA to
20mA per channel in order to reduce heat dissipation.
Here, the voltage gain of the inverting operational amplifier is dependent upon
the ratio between the input resistor, RINand its feedback resistor, Rƒ as
determined in the Op-amp tutorials.
A single 4-channel (Quad) SPST switch configured as a 4-to-1channel multiplexer
is connected in series with the resistors to select any feedback resistor to vary
the value of Rƒ. The combination of these resistors will determine the overall
voltage gain of the amplifier, (Av). Then the voltage gain of the amplifier can be
adjusted digitally by simply selecting the appropriate resistor combination.
Digital multiplexers are sometimes also referred to as “Data Selectors” as they
select the data to be sent to the output line and are commonly used in
communications or high-speed network switching circuits such as LAN´s and
Ethernet applications.
Some multiplexer IC´s have a single inverting buffer (NOT Gate) connected to
the output to give a positive logic output (logic “1”, HIGH) on one terminal and a
Page54
complimentary negative logic output (logic “0”, LOW) on another different
terminal.

Page55
It is possible to make simple multiplexer circuits from standard AND and OR
gates as we have seen above, but commonly multiplexers/data selectors are
available as standard i.c. packages such as the common TTL 74LS151 8-input
to 1 line multiplexer or the TTL 74LS153 Dual 4-input to 1 line multiplexer.
Multiplexer circuits with much higher number of inputs can be obtained by
cascading together two or more smaller devices.

The data distributor, known more commonly as a Demultiplexer or “Demuxer”

for short, is the exact opposite of the Multiplexer. The demultiplexer takes one
single input data line and then switches it to any one of a number of individual
output lines one at a time. The demultiplexer converts a serial data signal at
the input to a parallel data at its output lines as shown below. The circuit above
illustrates how to provide digitally controlled adjustable/variable op-amp gain
using a demultiplexer. The voltage gain of the inverting operational amplifier is
dependent upon the ratio between the input resistor, R INand its feedback
resistor, Rƒ as determined in the Op-amp tutorials.
The digitally controlled analogue switches of the demultiplexer select an input
resistor to vary the value of Rin. The combination of these resistors will
determine the overall voltage gain of the amplifier, (Av). Then the voltage gain of
the inverting operational amplifier can be adjusted digitally simply by selecting
the appropriate input resistor combination.
Standard Demultiplexer IC packages available are the TTL 74LS1381to 8-
output demultiplexer, the TTL 74LS139 Dual 1-to-4 output demultiplexer or the
CMOS CD4514 1-to-16 output demultiplexer.
Another type of demultiplexer is the 24-pin, 74LS154 which is a 4-bit to 16-line
demultiplexer/decoder.Heretheindividualoutputpositionsareselectedusinga4-bit
binary coded input. Like multiplexers, demultiplexers can also be cascaded
together to form higher order demultiplexers.
Unlike multiplexers which convert data from a single data line to multiple lines
and demultiplexers which convert multiple lines to a single data line, there are
devices available which convert data to and from multiple lines.

Procedure:
1. Connect the components as per circuit diagram.
2. Connect medical simulator as signal source.
3. Change SPST switch and check the output using multimeter /DSO.

Result:

Thus, analog multiplexer and demultiplexer were designed for bio signal.

Page56
Page57
Components of Photo transducer probe:

Infrared light absorption in the finger:

Page58
Expt.No:10
MEASUREMENT OF PULSE RATE USING PHOTO- TRANSDUCER
Date:

AIM: To measure the pulse rate using a photo transducer.

EQUIPMENT AND ACCESSORIES REQUIRED:

S. Name of the Equipment Quantity

No
1. Pulse rate monitor modules. 1
2. Sensor (finger probe) 1
3. Connectors. 1
4. DSO 1

THEORY

Pulse rate monitor is a non-invasive technique for the measurement of pulse rate
due to pulsatile blood flow. The pulse rate monitor determines the pulse rate by
passing two wavelengths of light, one red and one infrared, through the body
tissue to a photo-detector. The two wavelengths assumes that only two
absorbers are present; namely oxy hemoglobin (HbO2) and reduced hemoglobin
(Hb).

During measurement, the signal strength resulting from each light source
depends on the color and thickness of the body tissue, the probe placement, the
intensity of the light sources and the absorption of the arterial and venous blood
in the body tissues.

Pulse oximeter finger probe consists of two LEDs (Light Emitting Diodes), one
that transmits infrared light at a wavelength of approximately 940nm and the
other transmitting light at approximately 660nm. The absorption of these
selected wave lengths of light through living tissues is significantly different for
oxygenated hemoglobin (HbO2) and reduced hemoglobin (Hb) and is measured
with a photosensor.

The red and infrared LEDs within the probe are driven in different ways
depending on the manufacturer. Most probes have a single photodetector, so the
light sources are generally sequenced ON and OFF as shown in the diagram. The
output of the photodiode has raw signal as shown in the diagram.

There will be one signal that represents the absorption of red light (660nm) and
one that represents infrared light (940nm). The ac signal is due to the pulsating
of arterial blood while the dc signal is due to all the non- pulsing absorbers in
the tissue.

Page59
Page60
BLOCK DIAGRAM:

TABULATION:

S.NO. NAME OF THE SUBJECT PULSE RATE

WAVEFORM
(beats/min)

Page61
BLOCK DIAGRAM DESCRIPTION:

Pulse Amplifier: Pulse has to be amplified for further detection by electronic

circuitry, hence a pulse amplifier is required. The output voltage is 1V peak to
peak.
Pulse Detector: Pulse detect or gives a pulse after the detection of peripheral pulse.
The minimum
Input voltagerequiredis1Vand frequency is from 2Hz-20Hz.

Refra Generator: The refractory generator is a non-trigger able monostable

multivibrator which generates a 250ms delay following the pulse detection. The
input is a pulse of 0-5V (Min 1msec) and output is pulse of 0-5V(250msec).

Synch Generator: The synch generates a synchronous pulse with the incoming
wave. The input is a pulse of 0-5V(200msec) and output pulse is 0-5V(100msec).

F-V Converter: The frequency output of the synch generator is converted into
the voltage for detection of an increase or decrease of the heart rate. The input
pulse width is of 100msecand the output voltage is 1N/100 pulse per minute.

High Alarm and Low alarm: The modules high and low alarm are calibrated at
desired threshold level, e.g., pulse rate of 90pulses/min and 60 pulses/min
respectively for high and low alarm limit.

Audio Buzzer: The audio buzzer generates an audio beep when the heart rate
increases or decreases beyond the specified limit. The frequency of the buzzer
is1kHz and minimum voltage is 5V.

PROCEDURE
1. Connect the modules to form an instrument as in the block diagram.
2. Plug the sensor firmly in to the sensor port.
3. The finger to be inserted in the probe must be free from dirt or grease. Insert
the finger into finger probe.
4. Note the Pulse Rate (PR) interims of beats per minute from the monitor.
5. Drawtheplethysmographicwaveformofthesubjecttappedfromtheamplifierpoint
displayed on the DSO.

RESULT:
Thus, the pulse Rate is measured using a photo transducer.

Page62
Page63
pH Electrode–Combination type:

THE ELECTRODE SYSTEM FOR THE MEASUREMENT OF pH:

Where
A-Amplifier
D- Display
TC-Temperature compensation

Page64
Expt.No:11
MEASUREMENT OF pH AND CONDUCTIVITY
Date:
AIM
1. To measure the pH of the given bio logical samples.
2. Tomeasurethespecificconductivityofthegivenbiologicalsampleusingaconduc
tivity meter.

EQUIPMENTANDACCESSORIESREQUIRED

S. No Name of the Equipment Quantity

9. pH meter 1
10. pH electrode 1
11. Buffer solutions 1
12. Conductivity meter. 1
13. Conductivity cell. 1
14. Tissue paper &Distilled water As reqd.
15. Beaker, pipette, burette, flask and 1 each
THEORY:
pH-PRINCIPLE:
When one metal is brought in contact with another, a voltage difference occurs
due to their differences in electron mobility. When a metal is brought in contact
with a solution of salts or acids, a similar electric potential is caused, which has
led to the invention of batteries. Similarly, an electric potential develops when
one liquid is brought in contact with another one, but a membrane is needed to
keep such liquids apart. A pH meter measures essentially the electro-chemical
potential between a known liquid inside the glass electrode (membrane) and an
unknown liquid outside. Because the thin glass bulb allows mainly the agile and
small hydrogen ions to interact with the glass, the glass electrode measures the
electro- chemical potential of hydrogen ions or the potential of hydrogen. To
complete the electrical circuit, also a reference electrode is needed.
pH Meter:
The pH of a solution measures the degree of acidity or alkalinity relative to the
ionization of water. pH is defined as the negative logarithm of hydrogen ion
concentration. Measuring pH involves comparing the potential of solutions with
unknown [H+] to a known reference potential. pH meters convert the voltage
ratio between a reference half-cell and a sensing half-cell to pH values. Based
upon the Nernst equation, at 25°C, the output of a pH measuring electrode is
equal to 59.16 mV per pH unit. At 7.00 pH, which is the is potential point for a
perfect electrode, the output is 0 mV. As the solution pH increases (less acidic),
the mV potential becomes more negative. Conversely, as the solution pH
decreases (more acidic), the mV potential becomes more positive. At 25 C::pH 0
=+414.12 mV(Acidic)pH 4
=+177.480mV (Acidic)pH7=0.000mV (Neutral)pH9=-119.68mV (Basic)
Thevoltageontheouterglasssurfacechangesproportionallytochangesin[H+]. The pH
meter detects the change in potential and determines [H+] of the unknown by the
Nernst equation:
E=Er+(2.303RT/nF) log (unknown[H+]/internal[H+])
Where: E=totalpotentialdifference(measuredinmV)Er=referencepotentialR=gas
Page65
constantT=temperatureinKelvinn=numberofelectronsF=Faraday’sconstant

Page66
TABULATION:

S. SAMPLE pH VALUE mV in Temp

No. SOLUTIO pH
N Meter

General Information:
pH in body fluids pH
Gastric acid 0.7
lysosome 5.5
Granule of chromaffin cell 5.5
NeutralH2Oat37°C 6.81
Cytosol 7.2
CSF 7.3
Arterial blood plasma 7.4
Mitochondrial matrix 7.5
exocrine secretions of pancreas 8.1

Page67
GLASS pH ELECTRODE:
A glass pH electrode is the heart of pH meter. A glass electrode is a type ion
selective electrode made of a doped glass membrane that is sensitive to a
specific ion. A pH electrode consistsoftwohalf-
cells;anindicatingelectrodeandareferenceelectrode.Most applications today use a
combination electrode with both half-cells in one body. Over 90% of pH
measurement problems are related to the improper use, storage or selection.
Since pH glass electrodes measure H+ concentration relative to their reference
half-cells, they must be calibrated periodically to ensure accurate, repeatable
measurements. Although calibration against one buffer typically ensures
accurate pH readings, frequent two-buffer calibrations ensure them osteria able
results. Electrode should always be washed with distilled water and blot dried
with a paper towel. Electrode should be stored in electrode in storage solution to
ensure that your electrode glass stays hydrated.

BUFFER SOLUTION:
Buffers are solutions that have constant pH value and the ability to resist the
changes in that pH value. They are used to calibrate the pH measurement
system (electrode and meter).

PROCEDURE
CALIBRATION:
1. Switch the instrument ON.
2. Prepare buffer solutions of pH 4 and pH 9.2 by dissolving the respective
buffer tablets separately in 100ml of fresh distilled water.
3. Measure the temperature of the buffer solution and adjust the TEMP
4. COMPENSATECONTROLtothevalueofthetemperatureofthebuffersolutions.
5. Keep the container with4pHbuffersolution on the base plate of the electrode
stand.
6. Clip the electrode holding clamp at the appropriate height on the rod of
the electrode stand such that the electrode is immersed in the buffer
solution.
7. Push the pH /mV switch to pH position and push the STBY/READ switch to
READ position and adjust CAL control to set 4 on the READOUT and wait
for 30 seconds. Set the STBY/ READ switch to STBY position.
8. Removethecontainerwith4pHbuffersolution.
9. Wash the electrode with distilled water and blot clean with tissue paper.
10.SetSTBY/READswitchtoSTBYpositionandplacetheelectrodeinthepH9.2buffer.
11.A pH value of 9.2 will be
displayedontheREADOUT.Removethebeakerwith9.2pH buffer solution.
12. Clean the electrode with distilled water and blot clean with tissue paper.

pH MEASUREMENT OF GIVEN SAMPLE:

1. Measure the temperature of the given sample and set the
TEMPCOMPENSATE CONTROL and then immerse the electrode in the
given sample.

Page68
2. Set the pH/mV switch to pH position and STBY/READ switch to READ and
wait for 30seconds.
3. The pH value of the sample will be displayed on the READOUT.

Page69
 MEASUREMENT OF CONDUCTIVITY

Conductivity Meter:

PRINCIPLE:
Solution of electrolytes conducts electricity due to the presence of ions. The
specific conductance of a solution is proportional to the concentration of ionsinit.
In conductometric titrations conductance of the solution is measured during the
titration. Lowest conductance is usually observed at the end point. The titration
of HCl vs NaOH is given as an example. The reaction between HCl and NaOH is
represented as,

HCl+NaOHNaCl+H2O

When a solution of HCl is titrated with NaOH the fastmoving hydrogen ions are
progressively replaced by slow moving sodium ions resulting in the formation of
salt and water. As a result, conductance of the solution decreases. This decrease
will take place until the neutralization point is reached. Further addition of alkali
raises the conductance sharply, as there is an excess of hydroxide ions.
A graph is drawn between volume of NaOH added and the conductance of
solution. Two straight lines are obtained and the end point is indicated by
intersection of the two lines.

CONDUCTANCE:
The conductance ‘L’ of electricity through the solution depends upon the
migration of the cations and anions of the electrodes and different potentials. All
ionic species in a solution contribute to the ability of that solution to conduct
electricity, both intrinsic mobility and concentration of anions deciding the
Page70
extent of participation of each individual species. This property of the
electrolytic solution is known as conductance and is non

Page71
specific in nature. The conductance, which is a measure of the current that
results from the application of a given electrical force, is directly dependent
upon the number of charged particles in the solution.

The Conductance ‘L’ of a solution is inverse of resistance. Its unit is ohm -1or mho.
L=1/R
Specific conductance ‘k’: Since the conductance is reciprocal of resistance it
should be
proportionaltocrosssectionalareaandinverselyproportionaltothelengthofthe
conductor. Thus, L α A; L α 1/l
L=kA/l
k is proportionality constant called specific conductance. Its unit is ohm -1cm-
1or mho
cm-1

TABULATION:

S. SAMPLESOLUTION CONDUCTIVITYVALUE
No.

PROCEDURE
CALIBRATION:
1. After switching on the instrument, adjust the ‘SET 100’ control to read 100
on DPM and then calibrate the instrument.
2. After thoroughly cleaning, immerse the cell in standard solution,
say 0.1 N kcl (aqueous) which has specific conductivity of the order
0.01412 at 30◦C.
3. Put the SET/CAL/READ switch to CAL/READ position.
4. Select20mMHOrange.
5. After finding the temperature of the given solution with a thermometer,
Page72
refer the conductivity of standard solutions chart and adjust the CELL
CONSTANT to the corresponding value.

Page73
MEASUREMENT OF CONDUCTIVITY OF THE GIVEN SAMPLE:
1. Put the SET/CAL/ READ switch to SET position. Remove the conductivity
cell fromthe1N kcl solution and wash it with distilled water, blot with
tissue paper and immerse it in the unknown solution.
2. Select CAL/READ position and select appropriate range control and the
DPM will display the conductivity of unknown solution.

SAFETY PRECAUTIONS:
 Make sure that the electrode tip does not touch the beaker. This will lead
to breakage of the tip.
 Donotmixthestandardph7andph4solutionswithdistilledwaterinthebeaker.
This will alter the solution concentration and hence the pH values will
change
 Always ensure that the standby mode is selected before lifting the
electrode each time from the solution.
 Keeprepeatingthecalibrationwithstandardsolutionsph7,
ph4tillperfectvalueis obtained.
 Do not use the electrode beyond the pH of the temperature range.
 Every solution should be prepared by distilled water.
 Permanent damage may occur if used beyond the range.
 Electrode should be cleaned every time.

MAINTENANCE:
1. Conductivity cells should be kept immersed in water at 50c for few hours and
soaked in distilled water for at least 2 hours before use.
2. No air bubble should be sticking on to the plates

RESULT:
Thus, the pH value and conductivity value are noted and tabulated for
unknown solutions. pH value=
Voltage value= mv
Type of Solution=
Strength of the given HCl solution= N
Amount of HCl present in the given solution = g/l

Page74
Page75
Sphygmomanometer diagram:

Blood pressure measurement steps:

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Expt.No:12
MEASUREMENT OF BLOOD PRESSURE USING SPHYGMOMANO METER
Date:

Aim:
To measure blood pressure using sphygmomanometer.

Apparatus required:
S. No Name of the component Quantity
1 Analog/Digital sphygmomanometer 1
meter
2 Stethoscope 1
3 Handcuff 1

Theory:
A sphygmomanometer, also known as a blood pressure meter, blood pressure
monitor, or blood pressure gauge, is a device used to measure blood pressure,
composed of an inflatable cuff to collapse and then release the artery under the
cuff in a controlled manner, and a mercury or mechanical manometer to
measure the pressure. It is always used in conjunction with a means to
determine at what pressure blood flow is just starting, and at what pressure it is
unimpeded. Manual sphygmomanometers are used in conjunction with a
stethoscope. A sphygmomanometer consists of an inflatable cuff, a measuring
unit (the mercury manometer, or aneroid gauge), and a mechanism for inflation
which may be a manually operated bulb and valve or a pump operated
electrically.

Types:
Both manual and digital meters are currently employed, with different trade-offs
in accuracy versus convenience.

Manual:
A stethoscope is generally required for auscultation (see below). Manual meters
are used by trained practitioners, and, while it is possible to obtain a basic
reading through palpation alone, this only yields the systolic pressure.

Mercury sphygmomanometers are considered the gold standard. They show

blood pressure by affecting the height of a column of mercury, which does not
require recalibration. [2] Because of their accuracy, they are often used in
clinical trials of drugs and in clinical evaluations of high-risk patients, including
pregnant women. A wall mounted mercury sphygmomanometer is also known as
a Bau manometer.
Aneroid sphygmomanometers (mechanical types with a dial) are in common use;
they may require calibration checks, unlike mercury manometers. Aneroid
sphygmomanometers are considered safer than mercury sphygmomanometers,
although inexpensive ones are less accurate.[4] A major cause of departure from
calibration is mechanical jarring. Aneroids mounted on walls or stands are not
susceptible to this particular problem.

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Digital:
Digital meters employ oscillometric measurements and electronic calculations
rather than auscultation. They may use manual or automatic inflation, but both
types are electronic, easy to operate without training, and can be used in noisy
environments. They measure systolic and diastolic pressures by oscillometric
detection, employing either deformable

Page78
membranes that are measured using differential capacitance, or differential
piezo resistance, and they include a microprocessor. They accurately measure
mean blood pressure and pulse rate, while systolic and diastolic pressures are
obtained less accurately
thanwithmanualmeters,andcalibrationisalsoaconcernDigitaloscillometricmonitor
s may not be advisable for some patients, such as those suffering from
arteriosclerosis, arrhythmia, preeclampsia, pulse salter nans, and pulse us
paradoxus, as their calculations may not correct for these conditions,[citation
needed] and in these cases, an analog sphygmomanometer is preferable when
used by a trained person. Digital instruments may use a cuff placed, in order of
accuracy and inverse order of portability and convenience, around the upper
arm, the wrist, or a finger. Recently, a group of researchers at Michigan State
University developed a smart phone-based device that uses oscillometry to
estimate blood pressure. The oscillometric method of detection used gives blood
pressure readings that differ from those determined by auscultation, and vary
according to many factors, such as pulse pressure, heart rate and arterial
stiffness, although some instruments are claimed also to measure arterial
stiffness, and some can detect irregular heartbeats.

Procedure:
1. To begin blood pressure measurement, use a properly sized blood
pressure cuff. The length of the cuff's bladder should be at least
equal to 80% of the circumference of the upper arm.
2. Wrap the cuff around the upper arm with the cuff's lower edge one
inch above the antecubital fossa.
3. Lightly press the stethoscope's bell over the brachial artery just
below the cuff's edge. Some health care workers have difficulty
using the bell in the ante cubital fossa, some suggest using the
Bellor the diaphragm to measure the blood pressure.
4. Rapidly inflate the cuff to 180mmHg. Release air from the cuff at a
moderate rate (3mm/sec).
5. Listen with the stethoscope and simultaneously observe the
sphygmomanometer. The first knocking sound (Korotkoff) is the
subject's systolic pressure. When the knocking sound disappears,
that is the diastolic pressure (such as 120/80).
6. Record the pressure in both arms and note the difference; also
record the subject's position (supine), which arm was used, and the
cuff size (small, standard or large adult cuff).
7. If the subject's pressure is elevated, measure blood pressure two
additional times, waiting a few minutes between measurements.

Note: A BLOOD PRESSURE OF 180/120mmHg OR MORE REQUIRES IMMEDIATE

ATTENTION!

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Result:
The blood pressure measured by sphygmomanometer is found to be mm/Hg.

Page80
Block Diagram:

TABULATION

S. No Input Frequency (Hz) Blood flow velocity (cm/s)

Page81
Expt.No:13
MEASUREMENT AND RECORDING OF PERIPHERAL BLOOD FLOW
Date:

AIM
To measure the blood flow velocity using ultra sound transducer.

EQUIPMENTANDACCESSORIESREQUIRED
S. Name of the Equipment Quantity
No
1. Ultrasonic Blood flow meter 1
2. Ultrasound transducer 1
3. DSO 1

THEORY
Ultrasonic Blood flow meter is used to measure the velocity of blood flow.
Ultrasonic Blood flow meter are based on Doppler principle. An oscillator
operating at a frequency of 8MHz and the piezoelectric transducer is made of
barium titanate. The piezoelectric transducer is coupled to blood vessel and
ultrasonic beam with a frequency F is transmitted into the flowing blood. A small
part of the transmitted energy is scattered back and received by the transducer.
Since the scattering occurs mainly as a result of the moving blood cell, the
reflected signal has a different frequency due to the Doppler effect in frequency
may be (F+FD) or(F-FD), depending on the direction of the blood flow.
Using the Doppler Effect, it is possible to measure motions within the
body. The ultrasonic flow meter is a medical device that can approximate the
velocity of blood flow in a particular easel. The transducer of ultrasonic flow
meter is composed of both a transmitter
and a receiver component. Ultrasonic waves of a set frequency𝑓0are emitted by
the transmitter, which are then scattered by the moving red blood cells (RBCs)
that they come in contact with. These scattered waves return to the receiver
with a frequency altered by the
DopplerEffect(𝑓′). Figure1depictshowanultrasonicflowmeterfunctions.

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Page83
CALCULATION

𝒇(𝒄−𝒗𝒄𝒐
𝒇𝟏 𝒔𝜽)
= 𝒄

Where f1 = Apparent frequency =

8 MHz = Transmitted

frequency

c= Velocity of sound in blood=1570m/sec

𝜃=Angle of inclination of the in-accident wave to the direction of

blood flow V = Velocity of blood cell

Also, the incident & scattered radiation are both inclined at an angle 𝜃 to the
direction of flow, so

𝒇𝟐
𝒇𝟏𝑪
= (𝑪+𝑽𝒄𝒐
𝒔𝜽)

Doppler Shift

f =f -𝐟𝟐

Since c>>v

𝟐𝒗𝒇𝒗𝒄
∆𝒇= 𝒐𝒔𝜽
𝒄
𝒄∆𝒇
𝟐𝒇𝒄𝒐𝒔𝜽
V=

∆𝑓=Doppler shift frequency

Page84
RESULT:
Thus, the peripheral blood flow velocity is measured using ultrasound
transducer.

Page85
Expt.No:14
DESIGN A PCB LAYOUT FOR ANY BIO AMPLIFIER USING SUITABLE SOFTWARE
TOOL
Date:

AIM:

To create PCB layout of a given bio amplifier using software tool.

Apparatus Required:

S. No Name of the Component Quantity

1 PCB software tool 1

Theory:
Breadboards are great for prototyping circuits, but they aren’t so good for
actually using the thing you ’rebuilding. At some point, you ’probably
want to make a project more permanent. The best way to do that is to put
it on a PCB. The performance of your circuit will depend greatly on how
it’s laid out on the PCB, so I’ll give you lots of tips on how to optimize your
design.

You can always etch PCBs at home with a process that’s similar to
developing prints from photographic film. But that method is messy and it
uses a lot of chemicals. It’s much easier (and cheaper) to get your PCB
made by a professional manufacturer. To demonstrate the process, I’ll use
an online service called Easy EDA to design a PCB
layoutforanLM386audio amplifier, then I ’ll have it manufactured and
show you the results. Their free online design software is easy to use and
the rates are very affordable.

PCB Design Optimization:

Identify what each part of your circuit does, and divide the circuit into
sections according to function. For example, my LM386 audio amplifier
circuit has four main sections: a power supply, an audio input, the LM386,
and an audio output. It might help to draw some diagrams at this point to
help you visualize the design before you start laying it out. Keep the
components in each section grouped together in the same area of the PCB
to keep the conductive traces short. Long traces can pick up
electromagnetic radiation from other sources, which can cause
interference and noise.

The different sections of your circuit should be arranged so the path of

electrical current tis as linear as possible. The signals in your circuit
should flow in a direct path from one section to another, which will keep
the traces shorter. Each section of the circuit should be supplied power
with separate traces of equal length. This is called a star configuration,
and it ensures that each section gets an equal supply voltage. If sections
are connected in a daisy-chain configuration, the current drawn from
sections closer to the supply will create a voltage drop and result in lower
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voltages at sections further from the supply:

Page87
PCB Shape and Size:
It’s not uncommon to see round, triangular, or other interesting PCB
shapes. Most PCBs are designed to be as small as possible, but that’s not
necessary if your application doesn’t require it. If you plan on putting the
PCB into an enclosure, the dimensions may be limited by the size of the
housing. In that case, you’ll need to know the enclosure’s dimensions
before laying out the PCB so that everything fits inside. The components
you use will also have an effect on the size of the finished PCB. For
instance, surface mounted components are small and have a low profile,
so you’ll be able to make the PCB smaller. Through hole components are
larger, but they’re often easier to find and easier to solder.

User Interfaces:
The location of components like power connections, potentiometers,
LEDs, and audio jacks in your finished project will affect how your PCB is
laid out. Do you need an LED near a power switch to indicate that it’s on?
Or do you need to put a volume potentiometer next to a gain
potentiometer? For the best user experience, you might have to make
some compromises and design the rest of your PCB around the locations
of these components.

PCB Layers:
Larger circuits can be difficult to design on a single layer PCB because
it’s hard to route the traces without intersecting one another. You might
need to use two copper layers, with traces routed on both sides of the
PCB.

The traces on one layer can be connected to the other layer with avia. A
via is a copper plated hole in the PCB that electrically connects the top
layer to the bottom layer. You can also connect top and bottom traces at a
component’s through hole:

Page88
Page89
Ground Layers:
Some double layer PCBs have a ground layer, where the entire bottom
layer is covered with a copper plane connected to ground. The positive
traces are routed on top and connections to ground are made with
through holes or vias. Ground layers are good for circuits that are prone
to interference, because the large area of copper acts as a shield against
electromagnetic fields. They also help dissipate the heat generated by the
components.

Layer Thickness:
Most PCB manufacturers will let you order different layer thick nesses.
Copper weight is the term manufacturers use to describe the layer
thickness, and it’s measured in ounces. The thickness of a layer will affect
how much current can flow through the circuit without damaging the
traces. Trace width is another factor that affects how much current can
safely flow through the circuit (discussed below).To determine safe values
for width and thickness, you need to know the amperage that will flow
through the trace in question. Use an online trace width calculator to
determine the ideal trace thickness and width for given amperage.

PCB Traces:
If you look at a professionally designed PCB, you’ll probably notice that
most of the copper traces bend at 45° angles. One reason for this is that
45° angles shorten the electrical path between components compared to
90° angles. Another reason is that high speed logic signals can get
reflected off the back of the angle, causing interference:

If your project uses digital logic or high-speed communication protocols

above 200 MHz, you should probably avoid 90° angles and vias in your
traces. For slower speed circuits, 90° traces won’t have much of an effect
on the performance of your circuit.

Trace Width:
Like layer thickness, the width of your traces will affect how much current
can flow through your circuit without damaging the circuit.

The proximity of traces to components and adjacent traces will also

determine how wide your traces can be. If you’re designing a small PCB
Page90
with lots of traces and components, you might need to make the traces
narrow for everything to fit.

Page91
Procedure:

It all Starts with a Schematic

Before you start designing your PCB, it’s a good idea to make a schematic
of your circuit. The schematic will serve as a blueprint for laying out the
traces and
placingthecomponentsonthePCB.Plus,thePCBeditingsoftwarecanimportall
of the components, footprints, and wires into the PCB file, which will
make the design process easier (more on this later).Start by logging in to
Easy EDA, and create a new project: Once you’re on the Start page, click
on the “new Schematic” tab:

Now you ’ll see a blank can as where you can draw the schematic:

It’s best to place all of your schematic symbols on the canvas before
drawing any wires. In Easy EDA, schematic symbols are located in
“Libraries”. The default Easy EDA library has most of the common
symbols, butther earealso “User" Generated Libraries” with lots of other

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Page93
Each schematic symbol you use needs to have a PCB foot print associated
with it. The PCB footprint will define the component’s physical
dimensions and placement of the copper pads or through holes. Now is a
good time to decide which components you’ll be using.

The schematic symbols in the Easy EDA library already have footprints
associated with them, but they can be changed if your’ re using a different
size or style:

To change the footprint associated with a schematic symbol, search in the

“User Generated” libraries for a footprint that matches the component
you’re using. Once you find it, click on the heart icon to “Favorite” it:

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Page95
Then copy the name of the component:

Now click on the symbol in the schematic editor, and paste the name of
the new footprint into the “package” field in the right sidebar menu
(watch the video below for a demonstration):

Page96
Once all of your symbols are placed on the schematic and you’ve assigned
foot prints to each symbol, it’s time to start drawing the wires. Rather
than explain the details of all that in this article, I’ve made a video so you
can watch me draw the schematic for my LM386 audio amplifier:

After all the wiring is done, it’s a good idea to label the symbols. The
labels will be transferred over to the PCB layout and eventually be printed
on the finished PCB. Each symbol has a name (R1, R2, C1, C2etc.) and
value (10 μF,100Ω, etc.) that can be edited by clicking on the label.

The next step is to import the schematic into the PCB editor, but before
we do that, let’s talk about some things to keep in mind when designing
your PCB.

Creating the PCB Layout:

Now that we’ve discussed some off the ways you can optimize your PCB
design, let’s see how to layout a PCB in Easy EDA. Open your schematic
in the schematic editor, and click on the “Convert Project To PCB” button:

The footprints associated with each schematic symbol will be

Page97
automatically transferred to the PCB editor:

Page98
Notice the thin blue lines connecting the components. These are called
rats nest lines. Rats nest lines are virtual wires that represent the
connections between components. They show you where you need to
route the traces according to the wiring connections you created in your
schematic:

Now you can start arranging the components, keeping in mind the design
tips mentioned above. You might want to do some research to find out if
there are any special design requirements for your circuit. Some circuits
perform better with certain components in specific locations. For
example, in an LM386 amplifier circuit the power supply decoupling
capacitors need to be placed close to the chip to reduce noise.

After you’ve arranged all of the components, it’s time to start drawing the
traces. Use the rats nest wires as a rough guide for routing each trace.
However, they won’t always show you the best way to route the traces, so
it’s a good idea to refer back to your schematic to verify the correct
connections. Traces can also be routed automatically using the software’s
auto-router. For complicated circuits, it’s generally better to route traces
manually, but try the auto-router on simpler designs and see what it
comes up with. You can always adjust individual traces later. Now it’s
time to
definethesizeandshapeofthePCBoutline.Clickontheboardoutlineanddrag
each side until all of the components are inside: The last thing to do
before placing the order is to run a design rule check. A design rule check
will tell you if any components overlap or if traces are routed too close
together. The design rule check can be found by clicking the “Design
Manager” button in the right-side window:

Items that fail the design rule check will be listed below the “DRC Errors”
folder. If you click on one of the errors, the problem trace or component
will be highlighted in the PCB view. You can specify your own settings for
the design rule check by clicking the drop-down menu in the upper right-
hand corner and going to Miscellaneous> Design Rule Settings:

This will bring up a window where you can set design rules for trace
width, distance between traces, and other useful parameters:

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Page100
At this point it’s a good idea to double check your PCB layout against your
schematic to make sure that everything is connected properly. If you’re
satisfied with the result, the next step is to order the PCB.

Result:

Thus, the PCB layout of given bio amplifier is done using software tool.

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