Department of Bio-chemistry (JNMC)

SOPs

Dept. data last updated on :17/06/2026

General Laboratory Safety Procedures

DEPARTMENT OF BIOCHEMISTRY

JAWAHARLAL NEHRU MEDICAL COLLEGE

ALIGARH MUSLIM UNIVERSITY, ALIGARH

 

SOPs for General Laboratory Safety Procedures

 

DO

·         Know the potential hazards of the materials used in the laboratory. Review the Safety Data Sheet (SDS) and container label prior to using a chemical.

 

·         Know the location of safety equipment such as telephones, emergency call numbers, emergency showers, eyewashes, fire extinguishers, fire alarms, first aid kits, and spill kitswhichcanbefoundonallcampuses(IUPUIdoesnotprovidelaboratoryspillkits).

 

·         Review your laboratory’s emergency procedures with your Principal Investigator or Lab Supervisor to ensure that necessary supplies and equipment are available for responding to laboratory accidents.

 

·         Practice good housekeeping to minimize unsafe work conditions such as obstructed exits and safety equipment, cluttered benches and hoods, and accumulated chemical waste.

 

·         Wear the appropriate personal protective apparel for the chemicals you are working with. This includes eye protection, lab coat, gloves, and appropriate foot protection (no sandals or open toed shoes). Gloves must be made of a material known to be resistant to permeation by the chemical in use.

 

·         Shoes must cover the entire foot. Open toed shoes and sandals are inappropriate footwear in laboratories. Fabric and athletic shoes offer little or no protection from chemical spills. Leather shoes with slip-resistant soles are recommended.

 

·         Street clothing is to be chosen so as to minimize exposed skin below the neck. Long pants and shirts with sleeves are examples of appropriate clothing. Avoid rolled up sleeves. Shorts (including cargo shorts), capris and, miniskirts are inappropriate clothing in laboratories. Tank tops, sleeveless shirts and midriff-length shirts are not appropriate if not covered by a full-length laboratory coat and must not be worn if wearing an apron alone. Synthetic fabrics must be avoided in high-hazard areas where flammable liquids and reactive chemicals are utilized.

 

·         Contact lenses are not recommended but are permitted. Appropriate safety eyewear is still required for those that use contact lenses. Inform the lab supervisor of the use of contact lenses.

 

·         Wash skin promptly if contacted by any chemical, regardless of corrosively or toxicity.

 

·         Label all new chemical containers with the “date received” and “date opened.”

 

·         Labelandstorechemicalsproperly.Allchemicalcontainersmustbelabeledtoidentify the container contents (no abbreviations or formulas) and should identify hazard information.Chemicalsmustbestoredbyhazardgroupsandchemicalcompatibilities.

 

·         +Use break-resistant bottle carriers when transporting chemicals in glass containers that are greater than 500 milliliters. Use lab carts for multiple containers. Do not use unstable carts.

 

 

·         Use fume hoods when processes or experiments may result in the release of toxic or flammable vapors, fumes, or dusts.

 

·         Restrain and confine long hair and loose clothing. Pony tails and scarves used to control hair must not present a loose tail that could catch fire or get caught in moving parts of machinery.

 

DON’T

·         Eat, drink, chew gum, or apply cosmetics in rooms or laboratories where chemicals are used or stored.

 

·         Store food in laboratory refrigerators, ice chests, cold rooms, or ovens.

 

·         Drink water from laboratory water sources.

 

·         Use laboratory glassware to prepare or consume food.

 

·         Smell chemicals, taste chemicals, or pipette by mouth.

 

·         WorkaloneinthelaboratorywithoutpriorapprovalfromthePrincipalInvestigator or Lab Supervisor. Avoid chemical work or hazardous activities at night or during off-hours. Have a partner for assistance (use the “buddy-system”) at night or during off-hours.

 

·         Leave potentially hazardous experiments or operations unattended without prior approval from the Principal Investigator or Lab Supervisor. In such instances, the lights in  the laboratory should be left on and emergency phone numbers posted at the laboratory entrance.

 

 

 

 

 

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TISSUE CULTURE Lab of the Dept

DEPARTMENT OF BIOCHEMISTRY

JAWAHARLAL NEHRU MEDICAL COLLEGE

ALIGARH MUSLIM UNIVERSITY, ALIGARH


SOPs for TISSUE CULTURE LAB OF THE DEPARTMENT



Part-(A):- Personal Protective Equipment


  1. Lab coat or lab gown.

  2. Safety glasses if handling eye hazards.

  3. Gloves if handling risk group 2 microbes or clinical samples.

  4. Proper enclosed footwear 5. Hair tied back if long.


Part-(B):- Potential Hazards and Safety precautions

  1. Ultra violet lights in tissue culture room pose a burn hazard to skin, and blindness hazard to eyes. UV light can cause DNA damage leading to cancer. Ensure UV lights are off before starting work.

  2. Ethanol and other disinfectants can be toxic and/or flammable. Know the risks associated with each reagent before you start work.

  3. Tissue culture may pose a biohazard risk depending on what it is used for, e.g. when used to propagate viruses, this can be a serious human health risk. Know the risks associated with the biohazards before starting work.

  4. Tissue culture work is typically done inside a biosafety cabinet or other protective cabinet. Ensure you also read the correct risk assessment and SOP for the type of cabinet you are using – e.g. consult the SMB SOP for Biosafety II cabinets.

  5. The CO2 gas used in the tissue culture incubator can pose an asphyxiation or toxicity risk if inhaled. Ensure you have also read and understood the risk assessment and SOP for working with compressed gases. If you have any evidence that a CO2 leak is occurring or imminent, leave the room immediately and consult your supervisor and/or the room custodian.


Part-(C):- Procedure

  1. Check first with the room custodian before starting work. The custodian will instruct you on which hood and incubator to use – follow these instructions, and do not use other hoods/incubator.

  1. Ensure that the door to the tissue culture room is kept shut at all times. When in the tissue culture room, wear disposable gloves and a gown with long sleeves and cuffs. The gown is to be kept inside the tissue culture room and MUST NOT be used for general lab work. Do not wear your regular lab coat/gown in the tissue culture room – this can lead to contamination problems.

  2. Wipe down work area and equipment with 80% ethanol before starting.

  3. Everything used in the tissue culture lab must be labelled, including your initials, the date, and the cell line. For cell passages, it is usually best to include passage # and split ratio for your own information.

  4. If you have a lot of tubes, etc. that are not for long-term use, you can avoid labelling them all, but ONLY IF you dispose of properly immediately after you are done. At no time can you leave unlabeled materials in the lab.

  5. Turn the UV light on and fan on for 20 minutes before using the hood.

  6. Turn UV off before starting work, but leave fan on.

  7. UV irradiate for 20 minutes between cell types and strains and when finished.

  8. Everything going into hood must be swabbed with EtOH including gloved hands.

  9. Dry media bottles thoroughly if they have been taken out of the water bath (this water is a great source of contamination). Swab with EtOH, especially at the neck and bottom before placing in hood.

  10. Clean spills of culture or media immediately with EtOH.

  11. Ideally, hoods should be cleaned thoroughly approximately every 6 weeks, depending on usage. Swab interior with 70% ethanol. Wash the interior of the cabinet with disinfectant/detergent, including inside of glass panels, roof and walls. Swab interior again with 70% ethanol. Replace front panels and UV irradiate for a further 20 minutes.

  12. Regularly check the temperature and CO2 levels displayed on the incubator and also the CO2 levels of the cylinder, notify room custodian if temperatures are incorrect or CO2 levels are low.

  13. Swab gloved hands with EtOH before opening the incubator.

  14. Dispose of unwanted flasks appropriately ASAP, to minimize clutter and contamination risk (see Risk assessment & SOP ‘Disposal of biological wastes’).

  15. Clean spills of culture or media immediately with 70% ethanol– use the appropriate disinfectant for the biological agents present (e.g. ethanol is insufficient to kill viruses).

  16. All cell lines should be inspected regularly for microbial contamination.

  17. Check the water level (top up with autoclaved water only) and check for any contamination in the water bath for example turbidity or fungal growth.

  18. Water should be changed monthly (more often if heavy usage). Spray the inside of the bath with EtOH before refilling with autoclaved water.



Part-(D):- – Disposal

  1. Cell culture liquids must be disposed of by autoclaving, or treated freshly prepared 1% bleach. Anything that has been exposed to cell culture should also be similarly treated. After decontamination by these methods, liquids can then be disposed of via the sink.

  2. Do not leave waste media in the tissue culture room and do not over fill the biohazard bag. When the bag is 2/3 full, tie and take to be autoclaved on Level 2 and please replace the bag immediately!

  3. Any spills must be cleaned up immediately. Depending on the nature of the spill, clean up as described in SOPs for Biohazard Spills, Flammables, Corrosives, or Toxic Substances.

  4. Any large spills of hazardous materials (>1L) or incidents resulting in injury must be reported to your supervisor immediately and via the online incident report form within 24 h. Near misses (dangerous situations not leading to an incident) should also be reported.








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ELISA Reader

SOPs for ELISA Reader


1. PURPOSE

1.1 To determine the optimal concentration of a relevant capture antibody or soluble antigen.

1.2
To determine the optimal concentration of a lot of specific enzyme linked secondary antibody.

1.3
To determine the suitability for use of a lot of substrate buffer as measured against a lot of known performance.

1.4
To determine the suitability for use of a lot of PNPP substrate as measured against a lot of known performance.



2. SCOPE

2.1
To evaluate reagents to be used in quantitation of 12.8 antibody in solutions.

2.2
This assay can also be used to evaluate reagents for use in most ELISA procedures.


3. RESPONSIBILITY

3.1
All personnel performing optimization of ELISA reagents are responsible for adhering to this procedure.



4. REFERENCE AND APPLICABLE DOCUMENTS

4.1
TM-0060, ELISA for the quantitation of 12.8 antibody in solutions.


5. MATERIALS AND EQUIPMENT

5.1
See section 9.1


6. HEALTH AND SAFETY CONSIDERATIONS

6.1
See company safety manual.



7. DOCUMENTATION REQUIREMENTS

7.1
Laboratory Notebook 7.2 Attachment A. Reagent Qualification Form


8. GENERAL

8.1

New reagents should be qualified prior to use in an ELISA procedure.

8.2
By titering new or unknown reagents together with known reagents in a ELISA based assay a optimal concentration for use can be determined for the new component.

8.3
The suitability for use of new preparations can be determined by comparing new buffers or substrate preparations side by side with old preparations. The new preparations should give results similar or superior to the old lots.

8.4 ELISA plates should be covered during incubations to prevent evaporation and contamination.





9. PROCEDURE

9.1     Materials and Equipment

9.1.1
Microplate reader/recording spectrophotometer capable of reading @ 405 nm +/- 10 nm (Molecular Devices V-Max).

9.1.2  Multi-channelmicropipettes able to dispense up to 200 uL (8 Channel Finn pipette 50-300 ul).

9.1.3
Single channel micropipettes able to dispense up to 200 uL (Gilson 20-200 ul).

9.1.4     200 uL sized tips for micropipettes (Intermountain P-3200-1).

9.1.5     12 x 75 mm borosilicate tubes (VWR 60824-546).


9.1.6    15 mL conical tubes (Falcon 2095)

9.1.7     50 mL conical tubes (Falcon 2070)

9.1.8     1.0 mL serological pipets (Falcon 7520)

9.1.9     10 mL serological pipets (Falcon 7551)

9.1.10   Reagent troughs or square petri plates for use as reservoirs (Falcon 1003).


9.1.11
Polystyrene flat bottomed 96 well microtiter plate (Nunc maxisorb ELISA plates) 9.1.12    Vortex mixer (Vortex - Genie2) 9.1.13 ELISA plate Washer (Dynatech ultra wash 2)

9.1.14    Disposable sealing tape for 96 well plates (Corning #430454).


9.2       
Reagents

9.2.1     Dulbecco's Phosphate buffered saline (D-PBS without Ca ++ and Mg ++, JRH # 210-3025)

9.2.2   PT buffer = 0.5% tween 20 (Sigma # P-1379) in 1x D-PBS + 0.01% Thimerosal (Sigma T5125)

9.2.3  
PB buffer = 1.0% Bovine serum albumin (Miles Fraction V reagent grade # 210-3025)    In Phosphate buffered saline buffer + 0.01% Thimerosal (Sigma T5125).


9.2.4    AMP buffer 0.1 M 2-amino-2-mehtyl-1, 3-propanediol buffer, in 0.01% MgCl2 pH 10.3.

9.2.4.1
Dissolve 10.51 gm of 2-amino-2-methyl-1, 3-propanediol (Sigma # A9754) + 100 mg MgCl2 (Sigma M9272) per 1.0 L Deionized or distilled H2O + 0.01% Thimerosal (Sigma T5125). Make 5.0 litres.


9.2.4.2
Assign a lot number to this batch of buffer and assure that the number is written on each bottle along with the date and the initials of the technician preparing the buffer.


9.2.4.3
Each lot of buffer will be compared against the previous lot as described in this procedure. If this is the reagent to be qualified then samples of both the new and old lot will be needed.

9.2.4.4   Store the AMP buffer at 5 oC and make a new lot once/month.

9.2.4.5   Other substrates or buffer systems (e.g., HRPO) can be evaluated using this protocol. Follow instructions in SOP for particular assay for reagent preparations. 9.2.5 PNPP substrate solution - Dissolve 1.0 MG/ML p-Nitrophenyl phosphate (Sigma 104-100 or Zymed # 00-2201) in amino-2-methyl-1,3-propanediol buffer from 5.2.4. Each new lot of powdered PNPP will be qualified according to this procedure by comparing it against a previous lot. If this is the reagent to be qualified then samples of both the new and old lot will be needed. (Store PNPP at -20 ℃ (+/-5 ℃) in dark containers.)

9.2.6    Capture antibody or antigen used for coating plate. If this is the reagent to be qualified then samples of both the new and old lot will be needed.


9.2.7   Alkaline Phosphatase conjugated secondary antibody. If this is the reagent to be qualified then samples of both the new and old lot will be needed.

9.2.8    Bicarbonate buffer - 0.05 M pH 9.6 (Sigma # C-3041) make from powder; store at room temperature for up to 6 months.

9.2.9   Primary antibody or antigen for which the particular assay being evaluated is designed. If this is the reagent to be qualified then samples of both the new and old lot will be needed.

9.3      Evaluation of optimal concentrations of capture antibody (or antigen).

9.3.1
Prepare 1.5 mL of a 20 ug/mL solution of capture antibody or soluble antigen in Bicarbonate buffer.

9.3.2    Add 100 ul of bicarbonate buffer to each well of a 96 well ELISA plate.

9.3.3    Add 100 ul of coating material from 5.3.1 to each well of the first row of the ELISA plate. Titrate this material down the rows of the plate discarding the 100 ul of excessl material remaining from the last row.

9.3.4    Allow plate to incubate at 2-8 oC for 18-22 hr. 9.3.5 Wash the plates 3 times with PT buffer by sequentially dispensing >200 ul of buffer into the plate and aspirating off (repeated a total of three times using the plate washer).

9.3.6    Block the plates by adding 100 ul of PB buffer to each well.

9.3.7   Allow plates to incubate for 1.0 hour at R.T. and wash the plates with PT buffer as follows: 9.3.7.1 Wash 3 times as described in 5.3.5. Fill the plate with PT buffer.

9.3.7.2
Allow the plate to sit at R.T. for 5 minutes.

9.3.7.3
Wash 3 more times as described in 5.3.5. 9.3.8 Add 100 ul of PT buffer to each well of the plate.






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Flame photometer

SOPs for Flame Photometer


    The principle of flame photometer is based on the measurement of the emitted light intensity when a metal is introduced into the flame. The wavelength of the colour gives information about the element and the colour of the flame gives information about the amount of the element present in the sample.

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


Application


     The flame photometer is one of the most useful instruments in clinical analyses. This is due to the suitability of the flame photometer for determining sodium, potassium, and calcium, which are of immense importance in the development of the living being and indispensable to physiological functions. In the clinical analysis of sodium and potassium, the flame photometer gives, rapidly and accurately, numerous differential data for normal and pathological values.


Working Steps


·     Both the standard and sample solution are prepared in fresh distilled water.

·     The flame of the photometer is calibrated by adjusting the air and gas. Then the flame is allowed to stabilize.

·     Then switch on the instrument and the lids of the filter chamber are opened to insert appropriate colour filters.

·     The readings of the galvanometer are adjusted to zero by spraying distilled water into the flame.

·     The sensitivity is adjusted by spraying the most concentrated standard working solution into the flame. Now the full scale deflection of the galvanometer is recorded.

·     Again distilled water is sprayed into the flame to attain constant readings of galvanometer. Then the galvanometer is readjusted to zero.

·     Now each of the standard working solutions is sprayed into the flame for three times and the readings of galvanometer are recorded. After each spray, the apparatus must be thoroughly washed.

·     Finally, sample solution is sprayed into the flame for three times and the readings of galvanometer are recorded. After each spray, the apparatus must be thoroughly washed.

·     Calculate the mean of the galvanometer reading.

·     Plot the graph of concentration against the galvanometer reading to find out the concentration of the element in the sample.

·     The solvent is first aspirated to obtain fine solid particles.

·     These molecules in the solid particles are moved towards the flame to produce gaseous atoms and ions.

·     These ions absorb the energy from the flame get excited to high energy levels from the ground state.

·     But as these ions are unstable, they return back to ground state. While returning they emit characteristic radiation.

·     The intensity of emitted light is proportional to the concentration of the element.



Precautions


  • When using certain substances, the liberation of hazardous gases may require the use of a fume cupboard or other means of extraction.

  • Be aware of biological contamination. Ensure sterilization of the capillary tube, waste, nebuliser and mixing chamber is performed after contamination.

  • Ensure equipment is used on a clean, dry, non-combustible, solid work surface with at least 300mm suitable clearance all around andat least 1m clearance above the chimney.

  • Ensure that the outer chimney is properly installed before operatingthe equipment.

  • Ensure that the gas and air pipes have been connected correctly and that there are no gas leaks present before igniting.

  • Unit should be properly connected to the gas cylinder making sure there are no gas leaks.

  • Take care when fitting and removing the capillary tube to the nebulizer needle.

  • Do not leave the flame photometer unattended during operation.

  • Do not look down the flame chimney when igniting or operating the equipment.

  • Do not handle the flame chimney whilst in operation.

  • Do not lean or stretch over the equipment whilst in operation.

  • Do not cover the chimney whilst in use. Do not block or obstruct ventilation slots.

  • Do not spill substances onto the unit. If spillage does occur, disconnect unit from mains supply. If using hazardous substances

  • Allow sufficient time for the chimney to cool before handling.

  • Over adjustment of the fuel valve will cause excess flame.

  • Avoid installing the flame photometer in areas that are susceptible to drafts, but ensure that there is adequate ventilation to prevent any build-up of gas.

  • In order to completely disconnect power to the flame photometer the mains power cord must be detached from the back of the unit.

  • This hazard symbol is present on the flame chimney outer casing and indicates that the surface may be hot to touch.


Water bath

SOPs for Water bath


A water bath is laboratory equipment made from a container filled with heated water. It is used to incubate samples in water at a constant temperature over a long period of time. Most water baths have a digital or an analogue interface to allow users to set a desired temperature, some have their temperature controlled by a current passing through a reader


SOP (Standard Operating Procedure)


1.     Ensure that the platform is dry.

2.     Connect the power supply.

3.     Ensure the water level in water bath. It should be sufficient to pour the heating element.

4.     Switch “ON” the main power supply and instrument mains.

5.     Set the desired temperature of the water bath by rotating the temperature adjustment knob and indicator bulb shows red light until the set temperature is not achieved.

6.     The temperature sensor will maintain the set temperature during use of water bath.

7.     Switch “OFF” the instrument mains & main power supply after use.

1.     It is used in warming of reagents

2.     Melting of substrates

3.     Incubation of cell cultures.

4.     It is also enables certain chemical reactions to occur at high temperature.


Precautions


Glowing of Red lamp indicate mains “ON” & glowing of yellow lamp indicate heater “ON”








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Incubator

SOPs for Incubator


An incubator is a device used to grow and maintain microbiological cultures or cell cultures. The incubator maintains optimal temperature, humidity and other conditions such as the CO2 and oxygen content of the atmosphere inside. Incubators are essential for much experimental work in cell biology, microbiology and molecular biology and are used to culture both bacterial and eukaryotic cells.The most commonly used temperature both for bacteria such as the frequently used E. coli as well as for mammalian cells is approximately 37 °C (99 °F), as these organisms grow well under such conditions.


SOP (Standard Operating Procedure)


  1. Ensure that the incubator is properly connected to the power supply.

  2. Switch “ON” the main switch and then the cabinet switch, the indicator will glow green, red andgreen, respectively.

  3. Set the required temperature on digital display by pressing the set button along with arrow button upto increase the temperature/down to decrease the temperature.

  4. Observe the temperature shown on the digital display. The temperatureshould not differ by ±2°C.

  5. After achieving the desired temperature keep the samples inside the incubator on the tray and close the door firmly.

  6. Record the temperature of Incubator.

  7. Report any discrepancy observed during operation or temperature monitoring, to QC Manager.


Uses


Laboratory incubators provide a controlled, contaminant-free environment for safe, reliable work with cell and tissue cultures by regulating conditions such as temperature, humidity, and CO2. Microbiological incubators are used for the growth and storage of bacterial cultures.


Precautions

  1. Doorways and vents can blow in contaminants and increase the chance of fungal growth. What’s more, they can create drafts which can affect the temperature stability of your unit.

  2. Direct sunlight can cause temperature fluctuations and issues with anti-condensation features.

  3. We need enough space (at least three inches) around the unit so that heat can vent and cords and sockets are easily accessible.

  4. Place floor incubators on a stand to limit the risk of contaminants sweeping in when the door is open.

  5. Avoid humid, damp areas that may be harbouring fungal growth.

  6. Place units away from sources of vibration such as shakers, stirrers, or refrigerators, as vibrations can affect cell growth.

  7. Ensure the area around the unit is as clean as possible.


Colorimeter

SOPs for Colorimeter


Colorimetry is the measurement of the wavelength and the intensity of electromagnetic radiation in the visible region of the spectrum. Colorimetry can help find the concentration of substances, since the amount and colour of the light that is absorbed or transmitted depends on properties of the solution, including the concentration of particles in it. A colorimeter is an instrument that compares the amount of light getting through a solution with the amount that can get through a sample of pure solvent. A colorimeter contains a photocell is able to detect the amount of light which passes through the solution under investigation. The more light that hits the photocell, the higher the current it produces, hence showing the absorbance of light. A colorimeter takes 3 wideband readings along the visible spectrum to obtain a rough estimate of a colour sample. Pigments absorb light at different wavelengths.


SOP of Colorimeter


  1. Switch on the Colorimeter.

  2. Wait until the fluctuation will stop.

  3. Set the percentage transmission at zero.

  4. Take the Cuvette and clean it very properly. There should not be any trace on the surface of the Cuvette.

  5. Fill the sample in the Cuvette which we are going to measure.

  6. Place the Cuvette very slowly in the Cuvette chamber.

  7. Note down Optical Density of the given sample.

  8. Switch off the instrument.


Uses

1.      It is used by hospitals as well as laboratories for analysing biochemical samples such as urine, cerebrospinal fluids, plasma, biochemical samples, and serum. 

2.      It is widely used to generate a quantitative estimation of the serum components, proteins, glucose, and various biochemical compounds.

3.      It is also used in food industries and by manufacturing industries to make textiles and paints


Precautions

1.      This instrument is a precision measuring instrument.

2.      This instrument is not waterproof and should not be used in high humidity environment or water mist.

3.      Keep the instrument clean and tidy.

4.      If the instrument will not be used for a long time, remove the battery.

5.      The power adapter other than our company's special one cannot be used, otherwise the instrument may be burnt down.

6.      After the instrument is used, the colorimeter and the whiteboard cover should be placed in the instrument box and stored properly.

7.      The instrument should be stored in a dry, cool environment to avoid damage to the instrument.




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Office of the Department

The following staff members are responsible for smooth functioning of office work.


  • Mr. Syed Faisal Viqar

  • Mrs. Huma Shahid

  • Mr. Mohd. Faizan Ali


  • Receive & Dispatch:

All the paper received in the office by Mrs. Huma Shahid. 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 comment within three days.

The paper are their placed in the respective file folders (Office file/Personal file/Circular file/ Notice Board file/ PG file/NAAC/IQAC/File for Minutes of BOS).

Papers are then circulated/put on the Notice Board or given to concerned staff Teaching and Non-Teaching.

All papers, which are to be despatched from the office. The details are first entered in the despatch register.


  • Store

The following staff members are responsible for smooth functioning of store Mr. S. Nasir Ali (financial matters and matters related to Purchase).


  • Purchase of the following items are done

- Chemicals

- Equipments

- Departmental items (Fan, A/C, Furniture, Stationary etc).

All purchase/repair of the amount ranging between 1 – 25,000 – are done by Chairperson

Purchase ranging between 25,000 – 2.5 lakhs -  through the purchase Committee.

Purchase >2.5 lakhs are done through Finance office of the University.

  • Purchase of various items are done as follow / through the following processes to ask for quotations (at least four) are first asked by the concerned company/supplier.

The lowest cost quotation is considered, by the approval of Chairman.

Order is placed after Chairman approval.

It should be procured at the earliest possible time.

The received items are entered in the stock register and kept in the store.

Then depending upon the requirements of UG lab / research lab / PG lab / Clinical lab they are issued through the Indent book.


  • Indent book carries the following details:

Name of indenter

Name of items

Quantity


  • Indent book is then approved / permitted  / signed by the Chairman and the items are issued.

  • The following registers are maintained by the store keeper

Register for record of all the instruments in the department.

Register for Chemicals and glass ware.

Register for stationary.

Register for various items of the department

Register for record of repair of various items

Register for day to day expenses.

Miscellaneous

Budget Control Register

MAS Control Register

Register for consumables items

Register for Bank related work TA, DA.

Financial matters of Research Projects operated in the department are also taken care of by store keeper of the department.


  • Maintenance

Person concerned write an application related to the repair work and gives it in the
office of the Department.

Chairperson mark the application to the store incharge for necessary action.

Store incharge calls the person related to that particular work and ask Quotation for rates.

Chairperson  approved quotation of lowest rates.

Repair work is done at the earliest.

SOPs for Department in General


  • Official Working Hours 8:00 AM – 4:00 PM.

  • Research Scholars and MD students work in the research scholars even beyond these hours.

  • The concerned person will collect the keys from Chairman and open the department at 7:45 AM followed by opening of office labs, Chambers of the staff etc.

  • The extra lights switched on at night in the corridors, are switched off in the morning.

  • Sweepers are responsible for cleaning the department and ambience. Indoor plants and water cooler are taken care of by the non-teaching staff and the particular committee constituted.

  • All the staff members and students enter signature in the attendance register, in the displaying “on leave” if any is on leave.

  • Concerned staff update the (board name / designation board) and Notice board. Members of cleanliness and Ambience committee as well as members of the department take care of cleanliness.

  • Before closing the department all, the electrical switches and water taps are checked.

  • All the research scholars and MD students check the  electrical points and water taps before leaving the lab.


COVID-19

  • INSTRUCTIONS

Get Vaccinated

Keep physical distance of at least 1 meter from others.

Avoid crowd and close contact.

Wear a properly fitted  mask

Clean hands frequently with alcohol-based hand rub or soap and water.

Cover mouth and nose with a bent elbow or tissue when cough or sneeze.

Dispose off used tissues immediately and clean hands regularly.


  • Warning Signs for COVID 19

Fever

Dry cough

Headache

Tiredness

Dizziness

Breathlessness

Loss of smell and taste

Diarrhea


  • Seek Medical care immediately if any of these is present
  • If tested positive, isolate until recovery.

Clinical Lab

SOPs for Clinical Lab of the Department

 

v  Surface disinfection: lab bench is cleaned daily with any of the following Spirit- 70% alcohol or antiseptic/ Lysol/ Bleach (0.05%) solution. Preparation of bleach solution: 7gm or half tablespoon full per one litre of water 0.5%- 1:10 dilution; 0.05%- 1:100 dilution; 1% - 1:5 dilution, 0.1% -1:50 dilution)

 

v  The sample collection time is from8:30 am to 12:30 pm on weekdays and 8:30 am to 10:30 am on Friday.

 

v  For the analysis of Liver Function Tests (LFTs) 2 – 3 ml whole blood is required in serum vacutainer.

 

v  After collection, the samples are taken from collection desk to the clinical lab, where they are sorted and labelled.

 

v  Following this the particulars of the samples are entered in the lab register.

 

v  Samples are then centrifuged and serum is separated for performing the tests.

 

v  Haemolysed samples are discarded.

 

v  The tests for Bilirubin (direct and Indirect), AST, ALT, ALP and amylase are performed colorimetrically.

 

v  Calculations are done manually to obtain results.

 

v  The reporting is done following this on the investigation forms and the reports are also entered on the lab register for future reference.

 

v  The reports are available from 3:30 pm onwards from the collection counter itself.

 

v  Reagents are prepared on a weekly basis or as and when required.

 

v  Staff on duty follow the personal protective measures (use of gloves, mask, apron)

 

v  SR’s on duty supervise overall investigations and look for quality control.

v  Bio-medical waste generated is disposed off as per the guidelines

 

   

SOPs for Spill Management

 

1.0         Purpose

The purpose of this document is to provide guidance  during management of  accidental spills.

 

2.0         Scope

This document is use as a guidance document for management of  accidental spills for Clinical Biochemistry Laboratory, Department of Biochemistry, J.N.Medical College, AMU, Aligarh.

 

3.0         Roles and Responsibilities

3.1         Laboratory staff: It is responsibility of Mr. Ashfaq Ai, Mr. Raziuddin & Mr. Arsad Ahmad Laboratory technician to read and follow content of applicable SOP of Spill Management.

 

4.0         Procedure

4.1         Wear disposable gloves.

4.2          Cover spills with absorbent material and pour 1% disinfectant around  the spill and over the absorbent material and leave for 30 minutes.

4.3         And prohibit the place where spillage occurred by keeping a signage board.

4.4         Remove the absorbent material using brush and pan and place in the biohazard bags of infectious waste.

4.5         Wipe off the surface again with disinfectant.

4.6         Sweep broken glass etc. with a brush and collect in a dust pan. Discard into the sharps disposal unit/puncture proof container.

4.7         Report the spills to the laboratory In charge.

4.8         Keep a written record of such accidents.

 

5.0         Appendix

Appendix 1:Spill management log

 

6.0         References

6.1         NACO Manual

6.2         ISO 15189:2012

 

7.0         Appendix 1: Spill Management

 

Date

Spill

Reported By

Cleaned By

Checked by

 

 

 

 

 

 

 

 

 

 

 

 

SOPs for Biomedical Waste Management



8.0         Purpose

The purpose of this document is to provide guidance for proper segregation, treatment and disposal of biomedical waste generated by Clinical Biochemistry Laboratory  in a safe manner.

 

9.0         Scope

This document is used as a guidance document for proper segregation, treatment and disposal of biomedical waste generated by Clinical Biochemistry Laboratory, Department of Biochemistry, J.N. Medical College, AMU, Aligarh.

 

 

10.0     Roles and Responsibilities

10.1     Laboratory staff: It is responsibility of all laboratory staff to segregate the waste generated at the point of generation according to the SOP. 

10.2     Safai Karamchari: It is the responsibility of safai karamchari to take away the segregated biomedical waste daily

10.3     In Charge of Clinical Lab/Chairperson: In Charge of Clinical lab/Chairperson will ensure the compliance of the SOP.

    

10.4     Safety Issues and Waste Handling

10.4.1    Handle all the samples with care , as they can be potentially infectious.

10.4.2    Wear protective laboratory clothing’s in laboratory areas.

10.4.3    Do not pipette any reagent by mouth. Prevent splashing or spilling of samples or solutions containing samples. In case  of  spillage,  immediately  clean  it  with  1%  freshly  prepared  sodium hypochlorite. and dispose of the cleaning material by a suitable method.

10.4.4    Wash hands thoroughly with disinfectant after completion of the test.

 

10.5          Procedure  for segregation, transport and storage of biomedical waste  

10.5.1    Waste segregated at source as soon as it is generated. Refer to Appendix 1

10.5.2    Non-Infectious waste collected in black bags and disposed as general waste.

10.5.3    Infectious plastic is collected in labeled red bags after disinfection, sterilized by autoclaving and sent for disposal

10.5.4    Glassware disinfected by Sodium hypochlorite and sterilized by autoclaving  followed by  cleaning. Never attempt cleaning before Autoclaving.

10.5.5    The liquid infectious waste should be treated with chemical disinfectant for decontamination then neutralized and flushed  into sewer.

 

10.6          Document and Records

10.6.1    Laboratory technician maintain records related to the generation, collection, reception, storage, transportation, of bio-medical waste in accordance with the SOP and any guidelines issued.Refer to Appendix 1.

10.6.2    All records shall be subject to inspection and verification by the prescribed authority at any time.

10.6.3    Date and total number of each bag ,sterilization of Plastic waste and sharps container is recorded in a register and receipt of the same is maintained in the register.(Refer to  Appendix 03 & 04)

 

11.0          Appendix

11.1          Appendix 1: Table of Waste Segregation in color coded bags.

11.2          Appendix 2: Log of Disposal of BMWM Bags/Container.

11.3          Appendix 3: Log of Biomedical Waste Autoclaved

12.0     References

12.1     NACO Manual

12.2     ISO 15189:201

 

APPENDIX 1

Table Of Waste Segregation In Color Coded Bags

Color coding

Waste category

Red

Soiled disposable items like plastic syringes, Vacutainers, serum storage vials, used rapid test devices, used ELISA plates, micro tips; gloves, reagent vials

Yellow

All non sharps and non plastic infectious waste including human anatomical tissues,  cotton, filter paper, gauze contaminated with blood/ body fluids

White Puncture Proof

Needles, syringes with fixed needles, needles from needle tip cutter or burner, scalpels, blades, or any other contaminated sharp object that may cause puncture and cuts. This includes both used, discarded and contaminated metal sharps.

Blue

Broken or discarded and contaminated glass including

medicine vials and ampoules except those contaminated with

cytotoxic wastes. Body Implants

Black

general non infectious waste like paper, wrapper, cardboard box, kit boxes

 




















APPENDIX 2

LOG OF DISPOSAL OF BMWM BAGS/CONTAINER

 

Date

Time

No. of  BMWM Bags/Container

Signature of Safai karamchari

Signature of Lab technician

Red

Yellow

Black

White

Blue

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Incharge Signature…………………..

Date……………………….

 

 

 

APPENDIX 3

LOG OF BIOMEDICAL WASTE AUTOCLAVED

 

Date

Time Start

Time End

Temp & Pressure

Control tape

No.of Bags Autoclaved

Autoclaved by

Remark

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

  

Regents  for Estimation of  Alkaline Phosphatase:

 

1.

Carbonate Bicarbonate Buffer

=

Sodium bicarbonate (NaHCO3) = 1.68 gr

=

Sodium Carbonate (Na2CO3) = 3.15 gr

=

Dissolve in 500ml Distilled water

2.

Substrate

=

Phenylphosphate Disodium Salt Dihydrate (C6H5Na2O4P2H2O) = 1.09 gr

=

Dissolve in 500ml Distilled water

3.

0.5N NaOH

=

25 ml NaOH Solution 10N = Dissolve 500 ml distilled water

4.

Sodium bicarbonate 0.5N (NaHCO3)

=

Sodium Bicarbonate = 21 gr

 

 

=

Dissolve in 500 ml Distilled water

5.

4-Amino Antipyrine (C11H13N3O)

=

4 Amino Antipyrine = 3.0 gr

=

Dissolve 500 ml Distilled water

6.

Potassium Ferricynide K3Fe (CN)6

=

PotasiumFerricynide = 12 gr.

=

Dissolve 500 ml Distilled water

 

Preparation of STD

=

10ml Distilled water

                +

100 ml Phynol Solution

 

   

SGOT Aspartate  Aminotransferase (AST):

 

1.

Substrate

=

Aspartic Acid = 2.66 gr

=

 aKetoglutaric Acid = 30 mg

=

1.8 ml 10N NaOH

=

Makeup 100ml by stock Buffer

 

Stock Buffer = Potassium Dihydroge Phosphate = 2.7 gr

Phosphate Buffer = Disodium hydrogen = 11.3 gr

= Dissolve 1000 ml Distilled water

2.

2, 4 – Dinitrophenylhydrazine (DNPH) (C6H6N4O4)

=

DNPH = 100mg +50 ml HCl

=

Dissolve in 500 ml Distilled Water

3.

0.4 N NaOH

=

NaOH 10N = 80 ml

=

Dissolve in 2000 ml Distilled Water

= Dissolve in 1200 ml Distilled Water

 

Preparation of STD

Step (1)

=

=

Sodium pyruvate = 22 mg

10ml Phosphate Buffer

 

Step (2)

=

=

1ml stock Standard

4ml Phosphate Buffer

(Use 100 ml)

 

    

SGPT Serum glutamate – pyruvate transaminase (ALT):

 

1.

Substrate

=

DL Alanine = 1.8 gr

=

 aKetoglutaric Acid = 30 mg

=

18 ml Distilled water Dissolve

=

100 ml makeup by Phosphate Buffer

=

PH = 7.0

2.

DNPH

=

DNPH = 100mg

=

50 ml HCl

=

Dissolve in 500 ml Distilled Water

3.

0.4 N NaOH

=

NaOH 10N = 80 ml

=

Dissolve in 2000ml Distilled water

Preparation of STD:

 

Step (1)        (Stock Standard)

=

Sodium pyruvate = 22 mg

=

10 ml Phosphate Buffer

 

Step (2)        (Use 100 ml)

=

1 ml stock standard

=

4 ml phosphate Buffer

 

 

Reagents for Estimation of Serum Bilirubin:

 

Diazo A

Sulfanilic Acid

=

1.0 gr

Hydrocloric Acid HCl

=

15 ml

Dissolve in 1000 ml

=

Distilled Water

Diazo B

Sodium nitrite

=

500 mg

Dissolve in 100 ml

=

Distilled Water

Diazo Reagents

Diazo A 10 ml

=

mixed

 

  

Regents  for Estimation of  Serum Amylase:

 

1.

Buffered Starch Substrate (PH 7.0)

=

Dry anhydrous disodium hydrogen phosphate = 2.66 gr

=

Benzoic Acid = 800 mg.

 

Both reagents dissolve in 50 ml Distilled water. Bring to the boil. Mix 40 mg of soluble starch in 5-10 ml. Cold distilled water in a beaker and add it all to the boiling mixture, rinsing the beaker out with additional cold water. Continue boiling for 1 min. then cool to room temperature and dilute to 100 ml. Keep the solution at 40 and prepare freshly each month.

2.

Stock iodine Solution (0.1N)

=

Pure Sublimed iodine = 1.3 gr.

=

Potassium iodine = 2.4 gr.

=

Both reagents dissolve in 20 ml Distilled water and make to 100 ml Distilled water.

3.

Working iodine solution

=

Potassium fluoride = 5.0 gr.

 

Dissolve in 10ml Distilled water and make to 100ml with Distilled water. Store at 40 in a brown bottle.

 

 

Undergraduate Lab

  • Undergraduate lab is cleaned daily in morning by the assigned person.
  • Reagents/ chemicals for the practical class are prepared one day in advance.
  • Setting of the lab is done, all reagents and glass wares are kept at their respective places.
  • Lab is cleaned again after the practical class is over.
  • Glass wares are cleaned and kept at their respective places.

Tissue Culture Lab

Part-(A):- Personal Protective Equipment


  • Lab coat or lab gown.

  • Safety glasses if handling eye hazards.

  • Gloves if handling risk group 2 microbes or clinical samples.

  • Proper enclosed footwear 5. Hair tied back if long.


Part-(B):- Potential Hazards and Safety precautions


  • Ultra violet lights in tissue culture room pose a burn hazard to skin, and blindness hazard to eyes. UV light can cause DNA damage leading to cancer. Ensure UV lights are off before starting work.

  • Ethanol and other disinfectants can be toxic and/or flammable. Know the risks associated with each reagent before you start work.

  • Tissue culture may pose a biohazard risk depending on what it is used for, e.g. when used to propagate viruses, this can be a serious human health risk. Know the risks associated with the biohazards before starting work.

  • Tissue culture work is typically done inside a biosafety cabinet or other protective cabinet. Ensure you also read the correct risk assessment and SOP for the type of cabinet you are using – e.g. consult the SMB SOP for Biosafety II cabinets.

  • The CO2 gas used in the tissue culture incubator can pose an asphyxiation or toxicity risk if inhaled. Ensure you have also read and understood the risk assessment and SOP for working with compressed gases. If you have any evidence that a CO2 leak is occurring or imminent, leave the room immediately and consult your supervisor and/or the room custodian.


Part-(C):- Procedure


  • Check first with the room custodian before starting work. The custodian will instruct you on which hood and incubator to use – follow these instructions, and do not use other hoods/incubator.

  • Ensure that the door to the tissue culture room is kept shut at all times. When in the tissue culture room, wear disposable gloves and a gown with long sleeves and cuffs. The gown is to be kept inside the tissue culture room and MUST NOT be used for general lab work. Do not wear your regular lab coat/gown in the tissue culture room – this can lead to contamination problems.

  • Wipe down work area and equipment with 80% ethanol before starting.

  • Everything used in the tissue culture lab must be labelled, including your initials, the date, and the cell line. For cell passages, it is usually best to include passage # and split ratio for your own information.

  • If you have a lot of tubes, etc. that are not for long-term use, you can avoid labelling them all, but ONLY IF you dispose of properly immediately after you are done. At no time can you leave unlabeled materials in the lab.

  • Turn the UV light on and fan on for 20 minutes before using the hood.

  • Turn UV off before starting work, but leave fan on.

  • UV irradiate for 20 minutes between cell types and strains and when finished.

  • Everything going into hood must be swabbed with EtOH including gloved hands.

  • Dry media bottles thoroughly if they have been taken out of the water bath (this water is a great source of contamination). Swab with EtOH, especially at the neck and bottom before placing in hood.

  • Clean spills of culture or media immediately with EtOH.

  • Ideally, hoods should be cleaned thoroughly approximately every 6 weeks, depending on usage. Swab interior with 70% ethanol. Wash the interior of the cabinet with disinfectant/detergent, including inside of glass panels, roof and walls. Swab interior again with 70% ethanol. Replace front panels and UV irradiate for a further 20 minutes.

  • Regularly check the temperature and CO2 levels displayed on the incubator and also the CO2 levels of the cylinder, notify room custodian if temperatures are incorrect or CO2 levels are low.

  • Swab gloved hands with EtOH before opening the incubator.

  • Dispose of unwanted flasks appropriately ASAP, to minimize clutter and contamination risk (see Risk assessment & SOP ‘Disposal of biological wastes’).

  • Clean spills of culture or media immediately with 70% ethanol– use the appropriate disinfectant for the biological agents present (e.g. ethanol is insufficient to kill viruses).

  • All cell lines should be inspected regularly for microbial contamination.

  • Check the water level (top up with autoclaved water only) and check for any contamination in the water bath for example turbidity or fungal growth.

  • Water should be changed monthly (more often if heavy usage). Spray the inside of the bath with EtOH before refilling with autoclaved water.



Part-(D):- – Disposal


  • Cell culture liquids must be disposed of by autoclaving, or treated freshly prepared 1% bleach. Anything that has been exposed to cell culture should also be similarly treated. After decontamination by these methods, liquids can then be disposed of via the sink.

  • Do not leave waste media in the tissue culture room and do not over fill the biohazard bag. When the bag is 2/3 full, tie and take to be autoclaved on Level 2 and please replace the bag immediately!

  • Any spills must be cleaned up immediately. Depending on the nature of the spill, clean up as described in SOPs for Biohazard Spills, Flammables, Corrosives, or Toxic Substances.

  • Any large spills of hazardous materials (>1L) or incidents resulting in injury must be reported to your supervisor immediately and via the online incident report form within 24 h. Near misses (dangerous situations not leading to an incident) should also be reported.


General Laboratory Safety Procedures

DOs

·         Know the potential hazards of the materials used in the laboratory. Review the Safety Data Sheet (SDS) and container label prior to using a chemical.

 

·         Know the location of safety equipment such as telephones, emergency call numbers, emergency showers, eyewashes, fire extinguishers, fire alarms, first aid kits, and spill kitswhichcanbefoundonallcampuses(IUPUIdoesnotprovidelaboratoryspillkits).

 

·         Review your laboratory’s emergency procedures with your Principal Investigator or Lab Supervisor to ensure that necessary supplies and equipment are available for responding to laboratory accidents.

 

·         Practice good housekeeping to minimize unsafe work conditions such as obstructed exits and safety equipment, cluttered benches and hoods, and accumulated chemical waste.

 

·         Wear the appropriate personal protective apparel for the chemicals you are working with. This includes eye protection, lab coat, gloves, and appropriate foot protection (no sandals or open toed shoes). Gloves must be made of a material known to be resistant to permeation by the chemical in use.

 

·         Shoes must cover the entire foot. Open toed shoes and sandals are inappropriate footwear in laboratories. Fabric and athletic shoes offer little or no protection from chemical spills. Leather shoes with slip-resistant soles are recommended.

 

·         Street clothing is to be chosen so as to minimize exposed skin below the neck. Long pants and shirts with sleeves are examples of appropriate clothing. Avoid rolled up sleeves. Shorts (including cargo shorts), capris and, miniskirts are inappropriate clothing in laboratories. Tank tops, sleeveless shirts and midriff-length shirts are not appropriate if not covered by a full-length laboratory coat and must not be worn if wearing an apron alone. Synthetic fabrics must be avoided in high-hazard areas where flammable liquids and reactive chemicals are utilized.

 

·         Contact lenses are not recommended but are permitted. Appropriate safety eyewear is still required for those that use contact lenses. Inform the lab supervisor of the use of contact lenses.

 

·         Wash skin promptly if contacted by any chemical, regardless of corrosively or toxicity.

 

·         Label all new chemical containers with the “date received” and “date opened.”

 

·         Labelandstorechemicalsproperly.Allchemicalcontainersmustbelabeledtoidentify the container contents (no abbreviations or formulas) and should identify hazard information.Chemicalsmustbestoredbyhazardgroupsandchemicalcompatibilities.

 

·         +Use break-resistant bottle carriers when transporting chemicals in glass containers that are greater than 500 milliliters. Use lab carts for multiple containers. Do not use unstable carts.

 

·         Use fume hoods when processes or experiments may result in the release of toxic or flammable vapors, fumes, or dusts.

 

·         Restrain and confine long hair and loose clothing. Pony tails and scarves used to control hair must not present a loose tail that could catch fire or get caught in moving parts of machinery.

 

DON’Ts

·         Eat, drink, chew gum, or apply cosmetics in rooms or laboratories where chemicals are used or stored.

 

·         Store food in laboratory refrigerators, ice chests, cold rooms, or ovens.

 

·         Drink water from laboratory water sources.

 

·         Use laboratory glassware to prepare or consume food.

 

·         Smell chemicals, taste chemicals, or pipette by mouth.

 

·         WorkaloneinthelaboratorywithoutpriorapprovalfromthePrincipalInvestigator or Lab Supervisor. Avoid chemical work or hazardous activities at night or during off-hours. Have a partner for assistance (use the “buddy-system”) at night or during off-hours.

 

·         Leave potentially hazardous experiments or operations unattended without prior approval from the Principal Investigator or Lab Supervisor. In such instances, the lights in  the laboratory should be left on and emergency phone numbers posted at the laboratory entrance.

Safety Procedures In Research Laboratory

  • A science laboratory is a place where basic experimental skills are learnt only by performing a set of prescribed experiments. Safety procedure usually involves chemical hygiene plan and waste disposal procedures and significant physical and health hazards associated with the specific type of research and instruction in specific procedures that researchers should use in order to prevent and limit exposure to the health hazards in that workplace. Safety is a learned behavior that must incorporate into our instructional plans.


    v  PERSONAL PROTECTIVE EQUIPMENT AND CLOTHING

    Personal protective equipment is used in order to protect ourselves when working with chemical hazards. Common examples of personal protective equipment include: lab coats, footwear, gloves, safety goggles and glasses, face shields, hard hats and respirators.


    ·         Lab Coat

    The primary purpose of a lab coat is to protect against splashes and spills. A lab coat should be nonflammable and should be easily removed. Lab coats should be buttoned when in use. Rubber coated aprons can be worn to protect against chemical splashes and may be worn over a lab coat for additional protection.


    ·         Footwear

    Leather shoes which completely cover the toes, heel and top of foot provide the best general protection. The shoes must be made of water proof materials. The shoe must have a nonslip sole firmly attached to the foot. Sandals, sneakers, perforated shoes, open-toed shoes etc. do not provide adequate protection in case of spills.


    ·         Gloves

    When handling chemical, physical, or biological hazards that can enter the body through the skin, it is important to wear the proper protective gloves.


    ·         Eyewear

    Safety goggles provide the best protection against chemical splashes, vapors, dusts, and mists. Contact lenses should not be worn during any investigations using chemicals (even if you are wearing goggles).


    ·         Face Shields

    A face shield should be worn whenever there the entire face needs protection. Face shields can protect against impact, dust, particulates, and splashes to the face, eyes, and throat.


    v  EMERGENCY EQUIPMENT

    Research laboratory is equipped with a wide range of emergency equipment that can be invaluable in case of an accidental exposure to or a fire or explosion involving a hazardous reagent.


    ·         Fire Extinguisher

    Periodically check the date on the fire extinguisher to make sure that the extinguisher is full and the extinguisher is in good working order.


    ·         Eye Wash Stations

    Eye wash stations consist of a mirror and a set of bottles containing saline solution that can be used to flood the injured eye with water. The eye wash station is intended to allow us to flood the eye with a continuous stream of water.


    ·         Safety Showers

    Emergency shower are intended to provide on-the-spot cleansing when a chemical or solvent has been spilled.


    ·         First- Aid Kit

    A first aid must be readily available in science laboratory for use during accidents and emergencies. It is equipped with tincture of iodine, ferric chloride (alcoholic), burnol, savlon, cotton, sodium bicarbonate solution, adhesive plaster, bandages and scissors. Periodically inspect and restock the first aid kit.


    ·         Fume hood

    A fume hood is a type of local ventilation device that is designed to limit exposure to hazardous or noxious fumes, vapors or dusts.


    ·         Exhaust fans

    Exhaust fans must be fitted near the ceiling of laboratory for speedy removal of waste gases to keep the laboratory free from polluted air.


    v  HOUSEKEEPING

    The area is kept clean and items are stored in appropriate areas to ensure the safety of students. If everything is organized then labs will be more productive and cost effective.


    ·         Work area

    Work areas should be kept clean and free from unnecessary chemicals and obstructions. Reagents and chemicals to be used frequently are kept on reagent shelves. Reagent bottles are arranged in a definite order on these selves. All sensitive electronic equipment should be placed safely on table or within bag under table so that expensive damage can be avoided. We should not block access to emergency equipment (i.e. fire extinguishers, eyewashes, etc.), emergency shut-offs, and utility controls (i.e. electrical panels). When the fire alarm sounds we must evacuate the building via the nearest exit. Extinguish all flames and turn off all equipment before leaving.


    ·         Labeling samples and materials

    All containers containing chemicals or solutions of any kind that are retained between laboratory sessions must be labeled with full chemical name and hazard classification. The label must also contain the date and the name of the responsible person. Each laboratory must maintain chemical inventory that should be updated at least once in a year.


    ·         Chemical Handling and storage

    When dispensing chemicals, we should read the label carefully before starting the experiment. To avoid contamination and possible violent reaction we should never return unwanted chemicals to their container. We should not put dropper into a supply bottle. To avoid overheating and spurting never add water to concentric acid instead always add acid to water slowly with stirring. No pipetting should be done by mouth; we should use a pipette bulb or other pipetting device. Pipette must be used in a vertical position. Always dispense and dilute concentrated acid into a fume hood. Spilled chemicals should be cleaned up immediately and disposed of properly. Acids and  corrosive chemicals should be neutralized with soda ash (sodium carbonate) or sodium bicarbonate and spillage of alkali be neutralized by covering with dry sand. Chemicals should be stored in their original containers. We should not store chemical containers on the floor.


    ·         Hot equipment and glassware handling

    All glassware should be inspected before use and any broken, cracked, or chipped glassware should be disposed of in an appropriate container. All hot equipment should be allowed to cool before storing it. All glassware must be handled carefully and stored in its appropriate place after use. All glass chemical containers should be transported in rubber or polyethylene bottle carriers. Only borosilicate (Pyrax, Kimax etc) containers should be used for heating solutions. We should never heat a closed system such as a sealed test-tube or closed bottle. When a burner or hot plates are used we should always wear goggles and an apron to protect our eyes and clothing. We should never leave a hot plate unattended while it is turned on. Handle hot equipment with safety gloves and other appropriate aids but never with bare hands. We should keep our head, hands, hair, and clothing away from the flame or heating area, and turn heating devices off when they are not in use. Gas burners should be lit only with a spark lighter. Make sure all heating devices and gas valves are turned off before leaving the laboratory.


    v  UNAUTHORIZED ACTIVITY

    ·         Horse Play

    Horseplay in the lab is very dangerous. Unauthorized person should not allow entering in the laboratory.


    ·         Food, drink and Smoking

    No food or drinks of any kind are allowed in the laboratory. Any food or drink brought to the lab must remain in the carrying bag until they leave. We should not store food and drinks in laboratory refrigerators. Wash hands frequently throughout the day and before leaving the lab to avoid carrying toxic materials. Smoking is banned throughout the college and it is never allowed in any laboratories. In addition, do not apply cosmetics the laboratory. Never smell or taste chemicals or touch them with bare hands.


    v  WASTE DISPOSAL

    Use water taps only when required and should not waste any water. It is essential to clean the sink regularly. We should never put broken glass or ceramics in a regular waste container. Use a dustpan, a brush, and heavy gloves to carefully pick up broken pieces, and dispose of them in a container specifically provided for this purpose. Hazardous chemical waste including solvents, acids, and reagents should never be disposed of down sewer drains. Waste must be separated and disposed of in the proper waste containers.

Operation and Calibration of pH Meter

  • Objective:

To lay down the procedure to perform the Operation and calibration of pH meter

  • Key functions of pH meter

On/Off press the On/Off button of the pH meter to switch on the meter and switch off the meter.

CAL/MEAS:

HOLD/ENTER:

MI (memory input): Press MI to capture the measured reading of the pH with its corresponding temperature values and store them in memory.

Scroll through each SETUP and its sub group menu.

MR (memory recall): Press MR to retrieve the stored data from memory.

SETUP: Press SETUP  to activate the parameter setting menu to customize meter configuration, view calibration points and electrode offset data, select auto power off, reset meter and clear memory.

MODE: Select the measurement parameter option between pH with temperature.

  • General

Electrode Cleaning: Electrodes, which are mechanically intact, can often be restored to normal performance by one or combination of the following procedures.

Salt Deposits: Dissolve the deposit by immersing the electrode in raw water for ten to fifteen minutes. Then thoroughly rinse with de-ionized water.

Oil / Grease Films: Wash electrode pH bulb in a little detergent and water. Rinse electrode tip with de-ionized water.

Clogged Reference Junction: HeatHhhhhhhhhhHeat a dilute KCI solution to 60-80 C. Place the sensing portion of the pH electrode into the heated KCO solution for approximately 10 minutes. Allow the electrode to attain room temperature while immersed in some unheated KCI solution.

Electrode Activation: In case of dehydration/electrode showing sluggish response, the bulb can be re-hydrated by immersing the electrode in an ideal storage solution (e.g. buffer pH 5.00/5.01 solution) for 1-2 hours. If this fails, proceed for the electrode rejuvenation.

  • Operation of pH meter to determine pH of solution

Press ON to switch on pH meter. The MEAS annunciator appears on the top center of the LCD. The ATC indicate appears in the lower right hand corner to indicator automatic temperature compensation.

Rinse the electrode with de-ionized or distilled water before use to remove any impurities adhering to the probe body.

Maintain the temperature of the sample to 25C + 2C, unless otherwise specified in the individual monograph and dip the electrode along with the temperature sensor into the sample.

When dipping the electrode into the sample the sensor or the glass bulb of the electrode must be completely immersed into the sample, stir the probe gently in the sample to create a homogeneous sample.

Allow time for the reading to stabilize.

If the READY indicator has been activated the READY annunciator lights when the reading is stable

Press the HOLD to freezing of the measured reading and Press the ENTER to confirm the measured reading.

Record the observed values of pH and Temperature in the respective test data sheet or technical information sheet.

Wash the electrode in de-lonized water after use and store the electrode in storage solution or pH buffer 5.00/5.01 or 7.00/7.01 buffer solution.

  • Calibration of pH meter

Before starting the calibration make sure that the correct measurement mode is selected.

Wash the electrode thoroughly with de-lonized water or a rinse solution. Do not wipe the electrode, this causes a build-up of electrostatic charge on the glass suface.

Maintain the temperature of the buffers to 25C + 2C, unless otherwise specified in the individual monograph and dip the electrode along with the temperature sensor into the buffers. 

Perform the five point calibration buffer. The end of the electrode must be completely immersed into the sample. Stir the electrode gently to create a homogeneous sample.

Press CAL/MEAS key to enter pH calibration mode. The CAL indicator will be shown. The primary display will show the measured reading while the smaller secondary display will indicate the pH standard buffer solution reading.

Wait for the measured pH value to stabilize.

Press HOLD/ENTER key to confirm calibration. The meter is now calibrated to the current buffer.

Fist rinse the electrode with de-ionized water, which is followed by next buffer solution and place it in the buffer solution.

Follow steps 5.5.5 to 5.5.9 for additional calibration points.

When all the calibration points set in the unit configuration set up are completed the meter returns to the measurement mode automatically. However calibration can be terminated without completing the number of points as set in the unit configuration. It can be done by pressing CAL/MEAS to return to pH measurement mode. Record the calibration details and Temperature.

Change the calibration buffer every week or whenever required and record.

  • Determination of pH Electrode Slope

Press the SETP key to enter Set Up mode.

Press the MI/▲ or MR/▼ key to scroll through subgroups until you view parameter P3.0.

Press the HOLD/ENTER key. The display show the electrode offset value. If you have not calibrated at any buffer, the primary display show ‘-----‘

Press the HOLD/ENTER key to proceed to electrode slope display.

The display shows electrode slope in percentage. Slope displayed will be the current slope of the slope zone to which a measurement is made or calibration is done. Record the electrode slope.

  • Storage

Always keep the electrode bulb wet, preferably in the pH 5.00/5.01 buffer. Raw water is also acceptable storage media, but avoid storage in de-ionized water.

During handling of pH meter, check if electrode is broken, replace the same with new electrode and enter the details in instrument usage log book for pH meter.

  • Electrode Rejuvenation

If the electrode fail to give appropriate result during calibration, the same shall be Rejuvenated as mentioned below:-

Dip and stir the electrode in ethyl alcohol for 5 minutes.

Leave the electrode in tap water for 15 minutes.

Dip and stir the electrode in concentrated acid (e.g. Hydrochloric Acid, Sulphuric Acid) for 5 minutes

Leave the electrode in tap water for 15 minutes.

Dip and stir the electrode in strong base (Sodium Hydroxide) for 5 minutes.

Leave for 15 minutes in raw water.

Test with standard calibration buffer solution.

If the electrode still fails to restore the normal electrode response, the electrode rejuvenation steps (5.7.1.3 – 5.7.1.6) shall be repeated.

If the response does not improve, then the electrode has completed its useful life and same shall be replace with a new electrode.

Record the Electrode Rejuvenation details.

Electronic Balance

  • All balances should be located on a sturdy bench, free of vibration and drafts. The balance should be leveled. Adjust the position of the bubble in the level indicator by using the screw feet at the bottom of the instrument. The bubble should be in the center of the circle. Leave balance on at all times to maintain thermal equilibrium.

  • Clean balance after each use using a non-abrasive cleaner or a lint-free brush. Use only clean weighing vessels for weighing. Disposable, plastic weigh boats are most convenient for weighing of various chemicals.

  • Place weighing vessel (use smallest possible container) squarely in the center of the weighing pan.

  • Tare the instrument by pressing the re-zero or tare position on the control bar of the instrument. Instrument should read zero before adding substance. The weight of the container is now tared out.

  • With a clean scoop, add a small portion of sample at a time to avoid overshooting target weight. Avoid touching weigh vessel or pan. After reaching target weight, remove weigh vessel with sample and re-tare (re-zero) balance by pressing the tare (re-zero) position on the control bar.


Microscope

Procedure:

1.      Ensure that the microscope and its surrounding area is clean.

2.      Plug the microscope power cord in to electrical out let.

3.      Turn on the microscope by switching it on from main switch and two other switches one at back of microscope and at left lower side of microscope. This will switch on the light of microscope also.

4.    Set the intensity of light to the lowest setting using illumination control knob present at bottom in front side of microscope.

5.      Place the specimen slide on the stage.

6.      Rotate the nosepiece to move the objective (10X/20X/40X/60X) into working position.

7.   Change the position of nosepiece by using the knob present on left lower side to focus on the specimen. See through the eyepiece to do so

8.    Next, focus the specimen through the left eyepiece by turning the eye tube. Cover the right eyepiece while doing this and be sure to focus with the left eye tube only, without using the focusing knob.

9.      Observe the slide.


Cleaning and Maintenance

1.      Whenever lack of contrast, cloudiness or poor definition is encountered, Clean the lower magnification objectives and optical surfaces with a lint free cloth or lens tissue moistened (not wet) with ethanol.

2.      Avoid excessive use of solvent for cleaning.

3.      Cover the microscope always with dust cover, whenever the microscope is not in use.

4.      Wipe the bottom of Oil immersion lens of a fast absorbing tissue paper before and after using the lens.


Zeiss Optical Fluorescence Microscope

  • Materials allowed on the Microscope:

1.      No restriction on samples unless they are non-standard biological samples requiring level 3 or greater isolation rooms.

2.      For biological samples, prior approval from respective supervisor is required.

3.      Oil immersion microscopy can be carried out.

4.      It is better for the candidate to know the excitation frequency for the sample but it is not a mandate.


  • Authorization Procedure:

Authorization will be given by the chairperson. The candidate must have the understanding of the general components of the Microscope; i.e. the fluorescence source, the different filters, the magnification tool and other basics. An understanding of the data capturing software. Data storage and transfer is permitted using a CD/DVD. No USB drives allowed. Hands on experience using the standard sample is carried out a couple of times. Safe switching on' and 'switching off' of microscope.


  • Standard Operating Protocol:

The standard rule for the microscope's use is to complete one's work and leave the microscope back to its initial state; made ready for the next operator's use.

1.      When the Microscope is off, all the electrical switches must also be off. The eyepiece cover, the plastic encapsulation must be in the right place. The lever that helps switch from the camera to the eyepiece must at all times in a switched off mode be directed to the eyepiece.

2.      When using the microscope for normal fluorescence microscopy, the mains must be switched on first. The computer must not be switched on along with the microscope. First, the microscope's power module must be switched on, then the microscope and then the Fluorescence power source (Xcite). Fluoroarc is another module that sits just below the Microscope's power source. This is the secondary Fluorescence source. As long as the 'Xcite' source is functional, 'Fluoroarc' must always be kept off.

3.      Before turning ON the microscope, always check the log book for last user entry. If the fluorescence lamp was used in the past hour, allow it to cool. Also, report any abnormalities in the log book to the system owner.

4.      Before placing the sample on to the microscope table and before switching the system off, please ensure that the magnification has not been set to max or minimum; i.e. 100X or 1X.

5.      While switching off, the system must be left as found by the user before switching it on.


  • Negligence:

1.      If the last user for instance forgets to write proper details of his/her run in the log book

2.      If after use, the eye-piece cover or the microscope encapsulation has not been put on properly

3.      If the table-top has not been cleaned properly after the experiment.

4.      If the user does not operate the hardware or the software properly and is visible in the form of remnants after the experiment or not saving data appropriately thus putting other users' data at risk

5.      If the fluorescence lamp is left on. These components even though long lived, are very crucial to the operation of other experiments as well. Leaving it on results in unnecessary heating of the lamp not allowing the next user to use it.

6.      If the user does not take care of the lenses and does not report a mishap. This negligence might result in others loosing precious working hours.


This negligence put the system at absolute risk and shows direct negligence towards other users.


  • Manual

An electronic copy of the maul as provided by the manufacturer is available on the computer supporting the Microscope's data capture.


  • Logbook

The Logbook clocks in every activity by the authorized users.


  • First Level Tool Maintenance

The first level maintenance involves a weekly check of the vital part of the microscope namely,


  1. Eye piece, Lens - for dirt
    1. If dirt is found, the chairperson must be notified who will then schedule an appointment with the Zeiss engineer for a cleanup.
    2. The Zeiss instructed not to try and clean up any of the internal components because each one is extremely expensive.

  1. Fluorescence lamp
If the intensity of the lamp or any of its filters is seen to be different, it must be reported.

Cooling Centrifuge

Procedure:

Identify the speed and duration at which you wish to centrifuge samples PRIOR to using this instrument.


1.          Check the rotor you intend to use and be certain that the rotor is rated for the speed at which you would like to use it.

2.         If the rotor is not capable of being operated at the target speed, you will need to identify the rotor that is capable of being used at the desired speed, and then transfer your samples to a centrifuge tube that will fit and rebalance the samples, remembering to include the lids when balancing.

3.        Place the rotor in the centrifuge Check the name of the rotor and confirm the target speed.  Locate the correct lid for the selected rotor and place it beside the centrifuge.

4.     Place the samples into the rotor.  If the samples do not all have the same mass, place samples with the same mass on opposite sides of the rotor.

5.       Once the samples are loaded, check the rotor type again, confirm that you have the correct lid for the selected rotor and then screw the rotor lid onto the spindle.  The rotor lid set screw turns opposite to “normal” screws.

6.        Once the rotor lid is secured, close the centrifuge lid and set the desired temperature, the desired speed and the desired spin time.

7.        When everything is set, press the “Start” button and wait for the instrument to ramp up to the desired speed.

8.       Any large vibration in the instrument or irregular noise may indicate that something has gone wrong with the run.  Press the “Stop” button and move away from the instrument. Immediately contact a faculty member who can assist you BEFORE you open the lid to the centrifuge.

9.            If no problems are detected as the instrument reaches the desired speed, you may leave the area until the run is complete.

10.   When the run has completed and the rotor has come to a complete stop, you may unscrew the rotor lid and carefully remove your samples.

11.    Should a sample vial have leaked during the run, you MUST IMMEDIATELY clean the spill and dry the rotor. It is easy to miss spilled liquids in the bottom of the positions of the rotor, however, this liquid will cause an imbalance in the subsequent run that may lead to catastrophic failure.

12.     Remove the rotor from the centrifuge and place in the appropriate box adjacent to the unit.

13.     Should a sample vial have leaked during the run, you MUST IMMEDIATELY clean the spill and dry the rotor.

14.     It is easy to miss spilled liquids in the bottom of the positions of the rotor, however, this liquid will cause an imbalance in the subsequent run that may lead to catastrophic failure.

15.    Clean the area around the centrifuge with 70% ethanol (denatured) and wipe dry. This MUST be done following EVERY run.  Failure to clean up after yourself may ruin subsequent users’ experiments and/or facilitate the spread of contamination throughout the department.


Safety: Wear gloves throughout the procedure.  Do not leave the instrument.


Troubleshooting: Inform your supervisor or person responsible for that area of any issues


Operating and Cleaning of Distilled Water Unit

  • Objective

To lay down the procedure for Operating and cleaning of distilled water unit


  • Procedure

Connect the water inlet pipe to the tap.

Switch on the main supply

Keep an empty clean water container below the water outlet pipe to collect the distilled water.

Switch on the unit.

Collect about 500 ml of distilled water initially in the tank and discard it. Then collect the required amount of distilled water by operating the unit

Analyse the distilled water every week and report.


  • Cleaning of the unit

Dismantle the unit with the help of maintenance person

Clean thoroughly the storage container.

Put about 500 ml of dilute (5%) Hydrochloric Acid in the distillation unit.

Keep the Hydrochloric Acid in the unit for about 12 hours.

Wash away the acid with tap water very thoroughly.

Start the unit and collect the water to about 2 Lit. discard the collected water

Clean the unit every month.


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