8/3/2019 Exp. 1 - General Guidelines

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GENERAL GUIDELINES AND LAB HANDLING MEASURES

DATE:

EXP. No: 1

Good laboratory work will help you and the course a grand success. Follow the

following guide lines very strictly. These will protect you and your experiments.

1. No eating, drinking or smoking in the lab.

2. No storage of food in the lab.

3. No mouth pipetting. Use appropriate pipetting aids that are available.

4. Wear gloves for certain experiments.

5. Work cleanly and in organized manner. Wipe your tissue culture hood bench with

70% ethanol.

6. Most important, label everything you use with your initial, date and name of the

reagent, buffer or medium.

Procedure for handling cells and medium

1. Use sterile glassware and pipettes.

2. When you open a glass bottle, before and after, flame the mouth in a flame.

3. Flame the glass pipettes before and after the use.

4. Use bulbs to control pipettes. Do not mouths pipette. This is to protect both you

and the cells from contaminating. Never insert pipette which may have contacted

cells back into your stock bottle of medium. Use a fresh pipette. Thus, to change

the medium in a dish or TC flask procedure is as follows:

a. Prewarm medium and serum to 37°C in water bath.

  b. Wipe bottles with filter paper and transfer to the tissue culture hood

(which you should have been wiped with ethanol, equipped with pipettes,

 beakers for waste, etc.).

c. Open bottles, flame tops, replace caps loosely but take that they won’t fall

off.

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d. Transfer, desired quantity of serum to the medium, or mix in separate

(sterile) container. Use bulb and flamed pipette for transfer.

e. Move dishes of cells to the hood.

f. Using sterile, flamed, but NOT plugged, Pasteur pipette, aspirate the

medium. Flame the Pasteur pipette between dishes. Use a separate pipette

for different cells. Don’t use too many dishes at a time.

g. Using sterile, flamed pipette, transfer desired amount of medium and

serum to the dishes. Do not reuse the pipette.

h. Return dishes to incubator without shaking to avoid spill – over of 

medium.

i. Reflame tops of bottles and close tightly. Close pipette cans.

 j. Work in a desired tissue cultured area.

k. Wipe surfaces with ethanol before starting of experiments.

l. Wipe bottles dry (e.g. if they have been standing in a water bath) before

moving them into tissue culture hood.

m. Work cleanly, mopping spills immediately.

n. Discard used medium, and especially contaminated plates carefully.

Otherwise, they serve as source of contamination.

o. Label bottles and dishes with initials, cell type, date, etc. when you open a

new bottle of medium or serum, write the date on it and indicate how

much has been removed. If you add anything to it, indicate this on the

 bottle. If there is only a small amount left in a bottle, discard it.

  p. After you have finished, remove your belongings put them away or 

discard properly. Wipe the working surfaces with ethanol.

Equipments and supplies

A sterile hood is available for your use. Keep UV, lamp on for at least 10 mins

 before use. Turn off the UV lamp while you work.