DOW INTERNATIONAL

MEDICAL COLLEGE
DUHS

DEPARTMENT OF PHYSIOLOGY

FIRST PROFESSIONAL MBBS

LOCOMOTOR MODULE

PHYSIOLOGY PRACTICAL MANUAL

Prepared by
Dr. Talha Idrees
Dr. Maimoona Parveen

Reviewed and edited by

Prof. Sadiqa Syed
 Practical No. 1: Introduction of Power Lab and Lab Tutor

Objective:
 To overview the History of Power Lab
 To be familiar with the major hardware and software components of the Lab
Tutor system

Origin of Power Lab

 Karl Friedrich Wilhelm Ludwig was one of the greatest researchers and teachers
in the history of physiology.
 Ludwig had based his investigation and teaching of physiology on chemical and
physical laws, explaining all physiological processes on the basis of measurable and
experimentally demonstrable phenomena--not on some speculative "vital force." He
invented a number of devices and methods to carry out his scientific approach to
physiology, many of them related to his interest in circulation and respiration.

Origin of Power Lab

 In 1847 he devised the kymograph to prove that blood is moved by mechanical

forces, not the invisible "vital force." Ludwig's kymograph used a mercury manometer
tube and a revolving drum to graphically record blood pressure variations and other vital
signs. With later modifications, the kymograph became a standard tool for recording
results of experiments.

 The Power Lab technology is derived from equipment such as the Ludwig drum
kymograph which recorded data on a revolving drum. This was replaced by paper chart-
recorders, such as the Grass polygraph. Then came computer-based data acquisition
systems. One of the first of these was MacLab, the predecessor of the AD Instruments
PowerLab/LabTutor system, which you will be using in this Lab.

Yesterday and Today

1890s 1960s Today
Grass polygraph

Drum kymograph according to Ludwig

 Panel, displaying a section of recorded data
The PowerLab and a LabTutor

PowerLab and LabTutor

 LabTutor is an HTML-based software package, designed specifically for
laboratory teaching and used in conjunction with ADInstruments' PowerLab.

 LabTutor controls the sampling, digitizing and storage of experimental data, and
allows you to display, manipulates and analyzes them.

Data Acquisition
• First, a transducer converts the signal of interest (for example blood pressure or
body temperature) into an analog voltage, whose amplitude usually varies over time. This
in turn is monitored by the recording hardware, which can modify the signal by
amplification and filtering, processes called 'signal conditioning'. The resulting signal is
sampled at regular intervals and converted from analog to digital form before
transmission to the attached computer, where the sampled data is stored and displayed.

Example of Data Acquisition

Signal Conditioners
AD Instruments signal conditioners are software-controlled pre-amplifiers for use with
Power Lab data acquisition systems. The two types you are likely to see are Pods and
Front-ends. Once connected, the signal conditioners are automatically identified by the
Power Lab system, and all settings are stored when the data file is saved on your
computer.
 Signal Conditioner is explained below:
Types of signal conditioners you may use in this course

 Pod used for electro-oculography;

 Amplifier for measuring galvanic skin resistance;
 Bridge amplifier for connection to force and pressure transducers.

Signal Transducers
Virtually any transducer that generates an analog voltage between ±10V may be attached
to the PowerLab. Either directly or through one of the signal conditioners mentioned
above.

Stimulation of nerves and muscles

PowerLab also has the ability to produce an output voltage to induce a response in a
nerve or muscle.
This stimulating bar electrode connects to the Isolated Stimulator on the PowerLab.
Signal Transducers

Event markers
Some transducers may be used to mark an event, like this pushbutton switch.

Force or pressure transducers

Some transducers measure force like this hand dynamometer.
 Chemical properties
Chemical properties may also be measured such as conductivity, dissolved oxygen, and
pH.

Organization of LabTutor Experiments: Every LabTutor experiment is organized

in basically the same way.

 Every experiment begins with an introductory page. On this page there is a brief
introduction and a list of learning objectives.
 Each exercise page contains a LabTutor panel in which data is recorded.
 After each Exercise page, there is an Analysis page. Data that you recorded during the
exercise is available here for you to make measurements and complete any tables or graphs that
are required.
At the end of the experiment is the Report section. Any recordings that are required for your
report are reproduced here, along with tables or graphs that you have completed. This section also
contains questions that you can answer by typing into the spaces provided.

Front panel of Powerlab includes following:

1. Power indicator light.
2. Analog output connections
3. Analog inputs:
4. Isolated Stimulator status light:
5. Dual Bio Amp input
6. Isolated Stimulator outputs:
7. Isolated Stimulator switch:
8. Pod ports
9. Trigger input, may be used to start or stop a recording
event
 Practical No. 2: Characteristics of Skeletal muscle contraction
Objective: To Record the Simple Muscle Twitch (SMT), summation, tetanization and
muscle fatigue.
EXERCISE 1: SIMPLE MUSCLE TWITCH
Objective: To record and measure the muscular twitch response to nerve stimulation, and
investigate recruitment as the stimulus strength increases.

Prerequisite:
1. Power lab along with bridge pot,
2. Human subject (Volunteer)
3. Electrode cream
4. Stimulating bar electrode.

THEORY:
Motor unit
It is defined as group of muscle fibers innervated by a single somatic motor neuron.
Skeletal muscles must be stimulated by a nerve to contract (motor neuron). Individual
muscle fibers respond to isolated stimuli in an all or none fashion. By varying the number
of motor units contracting at a given time, the amount of tension generated by the whole
muscle can vary. Strength of contraction increases with increased number of motor units
a phenomenon called Recruitment. The strength of the contraction varies with the
strength of the stimulus applied.

Quality of stimulus
 Strength of stimulus- subminimal, minimal (threshold), submaximal, maximal
and supramaximal
 Duration of stimulus.
As stimulus strength is being increased, progressively more muscle fibers reach their
thresholds and contract. Thus, the change in tension is due to the number of contracting
muscle fibers, not a change in how much tension the individual fibers are generating.
Note that stimuli below the minimum strength needed to trigger any of the muscle fibers
to reach threshold and undergo an action potential (i.e., subthreshold stimuli) will not
trigger any contraction in the muscle.

Threshold is considered to be the level of stimulation required to trigger the smallest

measurable contraction resulting from the excitation and contraction of the first few
muscle fibers. If stimulus is increased above threshold into a range of stimulus intensities
called submaximal stimuli, contraction strength will increase with stimulus intensity as
progressively more and more muscle fibers in the muscle undergo contraction. Finally,
when stimulus strength is increased above a certain level (maximal) no further increase in
tension occurs, as all muscle fibers in the muscle are contracting.

Procedure

1. Have the volunteer place his or her hand as shown above, with the fingers under the
edge of the table and the edge of the thumb resting lightly on the pulse transducer.
2. Wipe the electrode cream from the subject's wrist.
3. Apply a small amount of electrode cream to the pads of the stimulating bar electrode.
4. Make the subject hold the electrode with the free hand firmly in place at the site for
stimulation of the ulnar nerve at the wrist. Ensure that the edge of the subject's thumb
is resting lightly on the transducer.
5. Ensure that the stimulus current is set to 1 mA.
6. Set the stimulator switch to ON.
7. Click Start.

Lab Tutor will stimulate, record for a fixed duration of 0.5 seconds and then stop
automatically.

8. Increase the stimulus current to 2.0 mA.

9. Click Start.
10. Continue to increase the stimulus current in 1 mA steps, clicking Start each time.
For most subjects, the threshold stimulus at which a response is first observed is in the
range 3-8 mA.

When you first see a response, add a comment to the recording,

noting the subject's name and the stimulus current used.

S.NO. PHASES STANDARD OBSERVED

VALUES VALUES
1. Latent Period (L.P) 0.01 sec.
2. Contraction Period 0.04 sec.
(C.P)
3. Relaxation Period 0.05 sec.
(R.P)
4. Height of
Contraction
Recruitment with increasing stimuli

Recruitment

1. Reduce the amplitude by 1 mA.

2. Click Start.
3. Increase the amplitude in 0.5 mA steps, clicking Start each time and adding a
comment, noting the current used.
4. Continue this until the response no longer increases.
For most subjects, this maximal stimulus is in the range 6-15 mA.

EXERCISE 2: SUMMATION AND TETANUS

Objective: To record the processes of summation of contraction and tetanization

THEORY:

Summation means adding together of individual twitch contractions to increase the

intensity of muscle contraction. It occurs in 2 ways:
1. By increasing the number of motor units contracting simultaneously “multiple
fiber summation
2. By increasing the frequency of contraction “frequency summation’ that can lead
to tetanization.
Tetanization: by increasing frequency, the contractions are added to one another; and
whole muscle contractions become smooth and continuous.

Now stimulate with paired pulses, and investigate how varying the interval between the
pulses affects the response.
Use the same setup as in Exercise 1.

Procedure
1.Turn the Stimulator switch ON. In the Stimulator panel, set the current to
5 mA greater than the maximal stimulus you determined in Exercise 2.
2. Check that the stimulus interval is set to 1000 ms.
3.Click Start.
4.LabTutor will automatically deliver the stimulus and stop recording
after a fixed duration of 3 seconds.
5.In the Stimulator panel decrease the stimulus interval to 500 ms, and click Start.
6. Repeat this for intervals 200 ms, 150 ms, 100 ms and 50 ms, noting the
values in comments as you did above.
7.When you have finished, turn the Stimulator switch OFF, ready for the next
exercise.

EXERCISE 3: MUSCLE FATIGUE

Objective: To measure the decline in maximal force during a sustained
contraction, and examine some properties of muscular fatigue
THEORY:
Prolonged strong contractions of a muscle leads to muscle fatigue. It results mainly from
inability of contractile and metabolic processes of the muscle fiber to continue supplying
the same work output

The grip force transducer should already be calibrated for the volunteer, as
described in the previous page.

Procedure
1. Allow the volunteer to view the computer screen.
2. Click Start.
3. Ask the volunteer to maintain 25% maximal grip strength while
watching the recorded trace.
4. After 20 seconds, tell the volunteer to relax.
5. Click Stop.
6. Wait for 30 seconds to allow recovery of muscle function.
7. Repeat steps 2-6 for contractions of 50%, 75% and 100% of
maximal grip strength.
8. Allow the volunteer to rest for two minutes.
9. Turn the volunteer away so that he or she cannot see the computer screen.
10. Click Start.
11. Ask the volunteer to produce a sustained maximal contraction.
12. After, 8 to 10 seconds, or when the force has obviously declined, instruct
them to try harder.
13. After a further 8 to 10 seconds, repeat the encouragement.
14. A few seconds later, ask the volunteer to relax.
15. Click Stop.
16. Click Start.
17. Ask the volunteer to produce a sustained maximal contraction. Every 8 to
10 seconds, allow the volunteer to relax very briefly (half a second), and then
return to maximal contraction.
18. After 30 to 40 seconds click Stop.
19. Allow the volunteer to use his or her other hand if gripping the
transducer has become painful. Turn the volunteer so that they can see the
computer screen.
20. Click Start.
21. Ask the volunteer to produce a 50% contraction while watching the trace.
22. After 10 seconds, press the Enter key to enter a comment (to mark the time).
23. Have the volunteer close his or her eyes, and attempt to maintain exactly
the same contraction force for the next 30 seconds.
24. After the elapsed time, the volunteer should open their eyes, and
adjust the contraction force back to 50%.
25. Click Stop. Examine the trace.
Analysis
- Locate the beginning of the recruitment recordings. Type into the first column of
the table the current delivered to produce each response.
- Move the cursor over the waveform and click on the peak of each response to
transfer its value to the Value panel.
- Drag the value to the appropriate cell in the table.

- As you enter the data, the Graph panel will graphically display the relationship
between stimulus current and response size.

- Note the stimulus current at which the response no longer increases. This is called
the 'maximal stimulus'.

Precaution:
1. Some exercises involve application of electrical shocks to
muscle through electrodes placed on the skin.

2. People who have cardiac pacemakers or who suffer from

neurological or cardiac disorders should not volunteer for such
exercises.

3. If the volunteer feels major discomfort during the exercises,

discontinue the exercise immediately and consult your
instructor.

OSPE Questions:

1. Describe main steps of muscle contraction.

2. State simple muscle twitch and its phases.
3. Define motor unit and recruitment phenomenon.
4. Which factors determine quality of stimulus?
5. List two type of summation of contraction
6. Define tetanization.
7. At which stimulus interval did you observe tetanus?
8. Explain the mechanism behind this phenomenon.
9. Define fatigue and its state its causes.
 Practical 3: Electromyography (EMG)
Objective No. 1: To stimulate the median nerve at the wrist and record muscle activity
from adductor policies brevis (a thumb muscle)

THEORY
The action potentials generated by contracting muscle alter the electrical charge in the
surrounding extracellular fluid. These electrical changes are conducted through body
fluids, and can be detected from the surface of the skin using electrodes applied to the
skin. A variety of instruments can detect the differences in charge between the electrodes,
amplify them, and generate recordings of these electrical changes called
electromyograms (EMGs).
EMGs records electrical activity in muscle to detect nerve damage. EMGs are used
diagnostically to detect damage to muscle or to the neural pathways responsible for
triggering muscle contractions.
It is a technique for evaluating and recording the electrical activity produced by skeletal
muscles. The signals can be analyzed to detect medical abnormalities, activation level,
and recruitment order or to analyze the biomechanics of human or animal movement.

TYPES OF EMG
There are two kinds of EMG in widespread use:
1. Surface EMG
2. Intramuscular (needle and fine-wire) EMG

PRINCIPLE
– Stimulate the median nerve at the wrist and record muscle activity from the abductor
pollicis brevis (a thumb muscle).
– 'Latency' is the time elapsed from the start of the stimulus pulse (the start of each
record) to the start of the evoked response.
– Note: you may see a very early deflection. This is the stimulus artifact and must be
ignored.

PROCEDURE
– Electromyograms
– Evoked EMG

1. Set the pulse current in the Isolated Stimulator box to 8 mA by clicking the
arrows or dragging the slider control. Recording will automatically stop after 0.05
seconds.
2. Click Start every time that you wish to stimulate. You should expect to see a
waveform that looks something like this.
3. Apply manual pressure to the back of the stimulus electrode to ensure that the
 nerve is stimulated and that the electrode doesn't move around during the
exercise.
4. Adjust the electrode to find the best position for stimulation as judged by the
amplitude of the response.
5. If you cannot get a response, increase the pulse current to 10 or even 12 mA. If
there is still no response try stimulating the ulnar nerve. (In some people, the
abductor pollicis brevis is innervated by the ulnar nerve instead of the median
nerve - an example of anatomical variation).
6. Once the electrode is optimally placed, increase the amplitude in 2 mA
increments. Record the responses until either you reach 20 mA or the response no
longer increases.
7. Turn the stimulator switch OFF.
8. Remove the stimulus electrode and mark with a pen the electrode indentation in
the skin nearest to the hand.

Analysis
-Use the scroll bar at the bottom of the LabTutor panel to review records recorded with
stimulation at the wrist.
-Measure the latency of a single waveform (the magnitude of the waveform is of no
consequence).
'Latency' is the time elapsed from the start of the stimulus pulse (the
start of each record) to the start of the evoked response. Note; you may
see a very early deflection. This is the stimulus artifact and must be
ignored.
Click at the point where the response begins.

ABNORMAL RESULTS OF EMG

EMG is used to diagnose diseases that generally may be classified into one of the
following categories:
1. Neuropathies
2. Neuromuscular junction diseases and
3. Myopathies
Abnormal results may be caused by the following medical conditions
Alcoholic neuropathy
Amyotrophic lateral sclerosis (ALS)
Axillary nerve dysfunction
Brachial plexopathy
Carpal tunnel syndrome
Cervical spondylosis
Dermatomyositis
Duchenne muscular dystrophy
Alcoholic neuropathy
Amyotrophic lateral sclerosis (ALS)
Axillary nerve dysfunction
-Brachial plexopathy
-Carpal tunnel syndrome
-Cervical spondylosis
-Dermatomyositis
-Duchene muscular dystrophy

Nerve Conduction Velocity (NCV)

Objective No.2: To record the nerve conduction velocity by stimulating the median
nerve at the wrist.

Prerequisite:
1. Power lab along with bridge pot,
2. Human subject(Volunteer)
3. Electrode cream
4. Stimulating bar electrode.
Theory:
A nerve conduction velocity (NCV) test determines how quickly electrical signals move
through a particular peripheral nerve or it measures how fast an electrical impulse moves
through a nerve.

PURPOSE OF CARRYING OUT NCV test

• Used in the diagnosis of nerve damage or nerve dysfunction.
• Plays a major role in evaluation of patients with neuromuscular disorders.
• Determine disorders of peripheral nervous system

The common disorders that can be diagnosed by nerve conduction studies are:
• Peripheral neuropathy
• Carpal tunnel syndrome
• Ulnar neuropathy
• Guillain-Barré syndrome
 • Facioscapulohumeral muscular dystrophy
• Spinal disc herniation

MOTOR NCS

 Motor NCS are performed by electrical stimulation of a peripheral nerve and

recording from a muscle supplied by this nerve.

 Latency: The time taken by the electrical impulse to travel from the stimulation
to the recording site. It is measured in milliseconds (ms).

You will measure responses evoked by nerve stimulation at the elbow.

The latency of these responses is longer than those evoked by
stimulation at the wrist. You will be able to calculate nerve conduction
velocity from the difference in latencies.
Electrode attachment:

1. Remove any watch, jewelry, etc. from your wrists.

2. Plug the five-lead Bio Amp cable into the Bio Amp socket on the PowerLab unit.
3. Plug the five color-coded lead wires into the Bio Amp cable, as shown.
4. Firmly attach the dry earth strap around your palm or wrist. The fuzzy side of the dry
earth strap needs to make full contact with the skin. Attach the green lead wire to the
earth strap. If the dry earth strap has a single connector lead wire it should be inserted
onto the pin nearest the Earth label.
5. If alcohol swabs are available, firmly swab the skin with them in each area where
electrodes will be placed. Lightly mark two small crosses on the skin overlying the biceps
muscle, in the position for the biceps recording electrodes, as shown. The crosses should
be 2-5 cm apart and aligned with the long axis of the arm. Lightly abrade the skin at these
areas with abrasive gel or a pad.
This is an essential step as it decreases the electrical resistance of the outer layer of skin
and ensures good electrical contact.
6.Prepare the skin over the triceps for attaching the electrodes as outlined in step 5 for the
biceps. The position for the triceps recording electrodes is shown in the figure.
Prepare the disposable ECG electrodes by removing the backing film. Place the
electrodes onto the skin over the crosses so they adhere well.
7. Plug the four shielded lead wires into the Bio Amp cable ports for positive and
negative, CH1 and CH2.
8. Snap the lead wires from CH1 on the Bio Amp cable onto the electrodes on the
subject's biceps. Snap the lead wires from CH2 on the Bio Amp cable onto the electrodes
on the subject's triceps. It does not matter which is positive and which is negative.
9. Check that all four electrodes and the dry earth strap are properly connected to the
volunteer and the Bio Amp cable before proceeding.
10. Make sure the PowerLab is connected and turned.

Setup:

- Position the Bar Stimulus Electrode on the medial aspect of the front of the elbow
as shown here. The electrode requires firmer pressure at the elbow than at the
wrist because the nerve is deeper in the tissues. The orientation of the electrode
should be the same as for wrist stimulation, with the red dot positioned closest to
the elbow.
- Turn the Stimulator switch ON.
Procedure:
1. Set the current in the Stimulator panel to 8 mA.
2. Click Start every time you wish to stimulate. Do this several times, using these
low-amplitude pulses to help to find the best position for the electrode.
3. If you cannot get a response, increase the stimulus current.
4. Once you have found the best position for the bar stimulus electrode, increase
the stimulus to 15-20 mA.
5. Click Start.
6. Repeat several times.
7. Turn the stimulator switch OFF.
8. Remove the stimulus electrode and mark with a pen the electrode indentation in
the skin nearest to the hand. Remove the other electrodes.

Analysis NCV

Analysis
1. Measure and record the distance between the marks at the
elbow and at the wrist. This is the distance between
stimulation sites.
2. Use the same steps as outlined for wrist stimulation to
measure the latency of a single waveform in the LabTutor
panel.
3. Record the latency value in the table.

The conduction velocity is calculated automatically in the table, using

the equation:
Velocity = Distance ÷ Time
The units of velocity are mm/ms or, equivalently, m/s.

Precaution:
1. Some exercises involve application of electrical shocks to muscle through electrodes
placed on the skin.
 2. People who have cardiac pacemakers or who suffer from neurological or cardiac
disorders should not volunteer for such exercises.
3. If the volunteer feels major discomfort during the exercises,
discontinue the exercise immediately and consult your instructor.

Questions
1. How do you vary the frequency, affecting the contraction force?
2. Which interval causes the greatest contraction?

Transfer the latency from the Value panel to the Latency (Wrist) column of the
table. In the next exercise you will stimulate at the elbow and again measure the
latency.

Median nerve conduction velocity

OSPE Questions:
1.Define EMG?
2.What is indication for an EMG? (an EMG exclude or detect muscle conditions due to muscular
or neurological disease)
3.Define nerve conduction velocity.
4.What is a normal nerve conduction velocity? (50-60 meters /second)
5.Which physiological factors affect NCV? (age, height, gender, temperature)
6.How NCV can be increased? ( Increasing diameter of nerve and Myelination)
7.What does decreased NCV mean? (nerve disease or abnormal pressure on nerve).
8.What are the signs of nerve damage? (numbness or tingling in hands or feet, glove and
stocking sensation, muscle weakness, sharp pain hands, arms, legs or feet, dropping objects held
in hands, a buzzing sensation that feels like a mild electrical shock.