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Cell Culture Contamination

The document provides steps for dealing with cell culture contamination: 1. Examine flasks microscopically for contamination and remove infected flasks. Clean contaminated flasks with sodium hypochlorite and incubators with sodium hypochlorite or isopropanol. 2. For persistent contamination, discard all flasks, aliquots of supplements and open bottles, and decontaminate cabinets by fumigation or cleaning.

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62 views1 page

Cell Culture Contamination

The document provides steps for dealing with cell culture contamination: 1. Examine flasks microscopically for contamination and remove infected flasks. Clean contaminated flasks with sodium hypochlorite and incubators with sodium hypochlorite or isopropanol. 2. For persistent contamination, discard all flasks, aliquots of supplements and open bottles, and decontaminate cabinets by fumigation or cleaning.

Uploaded by

jeevan george
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Dealing with cell culture contamination

1. Use the microscope to examine all tissue culture flasks for any contamination (tiny dots of bacteria or stings of
hyphae from fungi / mould). Remove all infected flasks into an appropriate laboratory where no tissue culture
occurs.

2. Half fill the contaminated flask with 10% sodium hypochlorite. Leave for 2 hours before rinsing down the sink
with copious amounts of water. Wipe the outside of all non-infected flasks with 2.5% sodium hypochlorite and 70%
isopropanol

3. Clean the CO2 incubator thoroughly, including the water tray, with 2.5% sodium hypochlorite. The sodium
hypochlorite should be left to soak for a maximum of 5 minutes and rinsed off with water for irrigation and
absorbent tissue (to prevent sodium hypochlorite corroding the metal of the cabinet). Spray incubator with 70%
isopropanol and wipe with dry tissues to remove any residual sodium hypochlorite and water.

4. Refill the water tray with 1 litre of water for irrigation and a suitable concentration of mild detergent / fungiside
commercially available for water trays and incubators. Return the tray to incubator.

5. Discard all culture medium prepared at the same time as the culture medium used in the infected flasks (refer to
Reagent Preparation Records for reagent number).

6. Wipe out the cabinet used with 2.5% sodium hypochlorite (refer to SOP Care and Maintenance of an Incubator).
The sodium hypochlorite should be left to soak for a maximum of 5 minutes and rinsed off with water for irrigation
and absorbent tissue (to prevent sodium hypochlorite corroding the metal of the cabinet). Spray the cabinet with
70% isopropanol and wipe with dry tissues to remove any residual sodium hypochlorite and water.

7. Put all gowns to laundry when cleaning has been completed. Use freshly laundered gowns when cleaning
precautions are complete.

Persistent contamination
We recommend the following must be performed in addition to the points described in the above section if
contamination occurs frequently (more than once in a week).

1. Discard all cell culture flasks.

2. Discard all aliquots of penicillin /streptomycin, glutamine and fetal bovine serum and any open bottles of water
for irrigation.

3. Decontaminate the Class I / II cabinet by formaldehyde fumigation if possible.

4. Decontaminate incubator by using the usual laboratory cabinet cleaning procedure including disinfectants.

www.abcam.com/technical

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