Department of Physiology
STANDARD OPERATIVE PROCEDURE
General SOP of the Department:
Official Working Hours 8:00 AM – 4:00 PM, Monday to Saturday except for Friday where the working hour is 8:00 AM to 12:30 PM.
for the job, will collect the keys from Chairman and open the department at 7:45 AM followed by the opening of the office, labs, Chambers of the staff, etc.
All the teaching and non-teaching members sign the departmental attendance register daily in the morning.
Members of the Department, teaching as well as non-teaching informs the Chairman before leaving the Department.
Members availing any leave, provide a piece of prior information to the Chairman by filling out the respective leave form and submitting it in the office.
In case of any unforeseen emergency, the leave information is relayed to the chairman’s office through e-mail or call. Later on, the required leave form is filled and submitted.
The members of the Department, teaching, and non-teaching carry out their academic, administrative duties and other assigned duties as per the defined protocols assigned by BOS.
The designated non-teaching staff concerned with the closing of the Department ensures that all the electrical equipment/appliances except for the laboratory deep freezers and refrigerators are switched off.
After Closing the department, the key is delivered to the Chairman.
SOP of the Office of the Chairman.
The concerned person designated, open the Office room and switch on the internet router enabling internet connectivity for the Department.
All the papers/notices/circulars are received in the office of the Department. Details of paper are entered in the receipt register.
Papers are shown to the chairman for his/her comment.
Chairman returns the paper with his/her comment and if needed mark the paper to the relevant person related to the work.
The paper is then placed in the respective file folders (Office file/ Personal file/ Circular file/ Notice Board file/ PG file/ NAAC/ IQAC/ File for Minutes of BOS etc).
Copies of the papers are then circulated/put on the Notice Board or given to concerned Teaching and Non-Teaching staff marked by the Chairman.
The details of all papers which are to be despatched from the office are first entered in the despatch register.
They are then despatched to the respective offices (Dean’s office/ Principal’s office/ various section of Registrar office etc.)
The office maintains all the concerned academic and administrative records.
SOP for Undergraduate Laboratory
Every student must wear an apron before entering their scheduled labs.
All students are required to maintaining the decorum of the laboratories. Eating, drinking, and cell phones usage are not allowed in the laboratories.
Students should check their laboratory schedule a day earlier, read up the relevant topic from the practical Physiology book and come prepared for the experiment.
Pay due attention to the practical demonstration given by your teacher before each experiment.
Always bring your practical manual to the laboratory.
Apparatus required for performing experiments will be delivered only after signing in the record register with the laboratory technician.
Check the apparatus being issued to you. If there is any breakage or damage to the apparatus, it must be reported to the lab technician.
Equipment’s should be placed in a proper manner & handled carefully.
For clinical physiology practical maintain proper etiquettes and ethics with the subject provided. Take consent and explain the procedure to the subject before performing a procedure.
Maintain a clean and hygienic work area
After completion of an experiment, complete the assignment given in the lab manual pertaining to the particular experiment and get it checked by the concerned teacher.
SOP of the Department library.
The Department has a well-stocked Seminar Library. It facilitates both faculty and students to use the books for academic purposes.
The departmental library is open for all faculties and UG students of the Department of Physiology during college hours.
The Library books are cataloged with Accession numbers for each, in line with the procedure followed by the Maulana Azad Library of the University.
All users are required to sign the register kept with the designated non-teaching staff appointed as a caretaker of the library.
Certain books like the Dissertation/Thesis cannot be taken outside the Department and should be returned the same day.
The faculty/students issuing the book are responsible for the proper maintenance of the book and should return the book in the condition it was issued.
In case of damage or loss of the book, information is to be provided to the Chairman who may then decide the proper procedure of refurbishing the lost/damaged book.
SOP ‘S OF LABS AND ADVANCED SCIENTIFIC EQUIPMENT
The lab is meant for facilitating patients’ care and research studies.
Patients referred from various clinical departments present to the lab for different neurophysiological tests viz., Nerve Conduction Study, Brainstem Evoked Response Audiometry, EEG, VEP, etc.
The patient and his/her attendants are advised to wear masks and sanitize his/her hands.
The procedure is done most probably on the same day but if there is any time issue or the patient load is high then, the date is given to the patient for the test.
The patient’s registration is done before undergoing the nerve conduction velocity test.
The patient’s history is taken before the procedure
Neurophysiology tests are charged services hence information about the cost of the test is also given prior to the procedure.
The amount is deposited in the revolving fund on the same day or the next day.
The patient will be asked to remove any clothing, jewelry, hairpins, eyeglasses, hearing aids, or other metal objects that may interfere with the procedure.
The patient will be asked to sit or lie down for the test.
A recording electrode will be attached to the skin over the nerve with a special paste and a stimulating electrode will be placed at a known distance away from the recording electrode.
The nerve will be stimulated by a mild and brief electrical shock given through the stimulating electrode.
The patient may experience minor discomfort for a few seconds.
NCV is not conducted for patients having pacemaker implantation.
The stimulation of the nerve and the detected response will be displayed on an oscilloscope (a monitor that displays electrical activity in the form of waves.
Patients come to neurophysiology lab, department of physiology from different OPD’s & ward’s like Medicine, Surgery, Plastic Surgery, CTVS, TB & Chest, Orthopaedics, Paediatrics, Ophthalmology, ENT, Psychiatry, Neurology, Endocrinology, Dermatology and Gynaecology, etc.
NeuroPerfect plus Machine EMG-2000 series (EMG/NCV/EP Machine)
Motor Nerve Conduction studies:
1. Nerve is stimulated and the compound muscle action potential is recorded by the surface electrodes.
2. The recording electrodes are placed with the help of adhesive tapes over skin on the surface of the muscle.
3. Active electrode is placed on the belly of the muscle.
4. Reference electrode is placed over the electrically inactive site like tendon.
5. Ground electrode is placed in between the stimulating and the recording electrodes (to get the zero voltage reference point).
6. The stimulating current/ voltage is increased gradually up to a point where the increase in the intensity do not produce any increment in the amplitude of compound muscle action potential.
7. The nerve is stimulated at more proximal site.
8. Stimulating nerve at distal and proximal site results in two CMAPs of similar shape and amplitude.
9. Latency is greater for proximal stimulation as compared to distal stimulation due to greater distance between stimulating and recording electrodes.
10. Difference in latency is time taken for faster nerve fibers to conduct between the two stimulation points.
a. Onset latency is time in msec from stimulus artifact to first negative deflection of CMAP.
b. MNCV (m/sec)= distance between the 2 stimulation sites (mm)/ [latency site 2- latency site 1(ms)]
i.e. MNCV (m/s) = D/(PL-DL), where, D is the distance in mm between proximal and distal stimulation.
Orthodromic: the nerve is stimulated distally and the recording is done in the physiological direction of the sensory conduction i.e. proximally
Antidromic: the nerve is stimulated proximally and the recording is done in reverse direction i.e. distally.
Latency is recorded from stimulus artifact to initial positive or subsequent negative peak.
Single stimulation site is used to measure the SNCV.
Calculation: SNCV (m/s)= Distance/ latency
1. Two channels BERA recording machine with an electrode box and headphones from Medicaid Neuroperfect is used to record the responses.
2. Connected to the computer via a USB cable.
3. Tones and clicks are produced at different intensities, at different rates and frequencies. Click sounds are produced by the transducer having a diaphragm that moves outwards (towards the eardrum producing condensation phase) and inward (away from the eardrum producing rarefaction).
4. The recordings are printed with the help of the attached printer.
5. Both earlobes, area behind ears and forehead are cleaned. Clean any oil and grease. This reduces the impedance (should be kept less than 5 ohms).
6. Placement of the electrodes:
· Right ear: connect to blue plug (side A)
· Left ear: connect to blue plug (side B)
· Low forehead: connect to the black plug
· High forehead: connect to red plug with Y adaptor
· Recording electrode Cz(at the vertex) placed midline at anterior fontanelle.
· Reference electrode (Ai) at ipsilateral ear lobule or mastoid process and contralateral ear lobule (Ac).
· Ground electrode (Fz) on the forehead.
7. Montage :
· Channel 1: Ai- Cz
· Channel 2: Ac-Cz
· Ground: Fz
8. Place the headphones.
9. Select the ear (right and left) with the help of the control panel and mouse button.
10. Set the machine:
· Recording sweep: 15-20 ms
· Low-frequency filter: 20-30 Hz
11. Apply broadband click sound stimulus to the test ear. Rarefaction phase stimulus is used.
12. 30-90 dB are used to determine the threshold response. The minimum intensity required to stimulate produce wave V is the hearing threshold. Electric signals generated by nerve fibers in the CNS are picked up by the surface electrodes, amplified and the signal averaging is done.
13. Two recordings are generated for reproducibility
1. Set the equipment
a. Sweep speed: 5-10 ms/div
Spontaneous activity: 50 μ V /division
MUPs: 200 μ V /division
c. Filter: 20-10,000 Hz
2. Duration of MUPs: measured at a gain of 100 μ V /division, sweep speed of 5 ms/div, and low filter of 2-3 Hz.
3. Cooperative patient
4. Clean the surface, remove any oil, grease.
5. Electric interference should not be there
6. Surface electrodes
7. Select the muscle
8. Proper instructions to patients related to contraction and relaxation of the selected muscle.
9. Place electrode slightly away from the motor point to avoid the endplate noise.
10. Sharp MUPs on minimal contraction ensure the correct placement.
11. Record the activity of the muscle:
a. Spontaneous: originate from the neuromuscular junction or the terminal axon.
b. MUP on voluntary activity:
i. Recruitment pattern
ii. Motor Unit Action Potential
12. Remove the electrodes and clean the area.
13. Interpretation of the findings
Enzyme-Linked Immunosorbent Assay (ELISA) captures the target antigen/ antibody in the sample by utilizing the specific antibody/antigen and also helps to detect/ quantify the target molecule with the help of the reaction of the enzyme with its substrate. The enzymatic activity is measured calorimetrically.
a. To each well of 96 well ELISA microplate add diluted antibody.
b. To prevent evaporation seal the microplate.
c. Incubate at 40 C for at least 15-18 hours.
d. Remove the diluted antibody and wash at least three times.
a. To each well of the microplate add the buffer.
b. Incubate at 370 C for one hour.
c. Remove the buffer by washing at least three times.
3. Diluted samples are added to each well (100 μL)
4. Prepare the dilution series of the standard on the same microplate.
5. Incubate at 370 C for one hour.
6. Wash at least five times
7. Add the detection antibody:
a. Add 100 μL of the diluted detection antibody in each well of the microplate
b. Incubate at 370 C for one hour
c. The reaction occurs and now remove the detection antibody
d. Wash at least five times
8. Add the enzyme-labeled secondary antibody:
a. Add 100 μL of the diluted enzyme-labeled secondary antibody in each well of the microplate.
b. Incubate at 370 C for one hour
c. The reaction occurs
d. Remove secondary antibody
e. Wash at least five times
9. Add substrate solution:
a. Add substrate solution
c. Color develops
d. When sufficient color develops then add the stop solution
10. Measure the absorption with the help of a plate reader
Types of ELISA:
Target protein( or antibody) immobilized on the surface of microplate wells
Incubate with enzyme-labeled antibody to the target protein (or specific antigen to target antibody).
Measure the activity of the well-bound enzyme
Target protein immobilized on the surface of microplate wells
Incubate with an antibody to the target protein (primary antibody)
Incubate with secondary antibody against the primary antibody.
Measure the activity of the well-bound enzyme
Antibody to a target protein is immobilized on the surface of the microplate wells.
Incubate: Initially with the target protein and then with another target protein-specific antibody (labeled with enzyme).
Different epitopes of the target protein are recognized by immobilized and enzyme-linked antibodies.
Measure the activity of the well-bound enzyme.
Antibody specific for a target protein is immobilized on the surface of microplate wells.
Incubate with a sample containing the target protein and known amount of enzyme-labeled target protein.
Measure the activity of the well-bound enzyme.