There are different ways to disinfect the tissue culture media. Some are effective while others are not. In this blog, we will be discussing a standard operating procedure to disinfect tissue culture media in vacuum cell culture flasks.
This procedure requires you to utilize proper engineering controls and personal protective equipment. It also requires a list of materials to set up a vacuum flask and to ensure proper disposal and disinfection of tissue culture wastes. You would require a plastic vacuum flask, thick-walled plastic tubing, plastic or glass tube, HEPA filter, and rubber stopper. (blink, 2021)
Bleach is the best and easily accessible disinfectant for the chemical disinfection of biohazardous liquid waste. For every part of bleach, there should be nine parts of liquid waste and 30 minutes of contact time. This measurement and timing should be extremely accurate in order to carry out the perfect procedure. After the 30 minutes are over, the waste should also be sewered.
Here is how you carry out the perfect, error-free procedure to disinfect tissue culture media in a vacuum flask.
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Prepare The Primary Vacuum Flask With Bleach
The flask should have a clear level indicating the maximum fill. It is important to make sure all of your measurements lie under it to carry out this step without any errors. Your first step would be to prepare the vacuum flask with bleach before adding the tissue culture media. In order to disinfect the liquid waste, you need to add an adequate amount of household bleach. You want to create a final bleach concentration that is less than or equal to 10%. Always remember that you need only one part of bleach for nine parts of liquid waste. For example, you can make a mixture by adding 100 millilitres of bleach for every 900 millilitres of biohazardous liquid waste. Make sure that the vacuum flask doesn’t contain a volume that is more than two-thirds full. Furthermore, if you don’t have access to a biosafety cabinet, vacuum flasks should be stored inside a leak-proof and unbreakable secondary container that has sufficient space to hold the liquid waste.
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Label The Flask
It is important that you properly label the flask in order to warn other people who may be using the lab. Additionally, since the liquid waste is biohazardous, you should be especially careful when labelling it.
So apply the label that clearly states the content inside it is biohazardous and also apply a label that says, “Tissue culture media disinfected with bleach (9:1)”. You can use a separate label for the caution and contents or a combined label that indicates both biohazard warning and contents.
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Protect the Vaccum System With A Filter
This can be achieved by inserting an in-line hydrophobic HEPA filter before the flasks’ vacuum outlet that is contracted to the vacuum system of the building. If you have individual vacuum pumps, HEPA filters are not required. Although due to the danger of contamination of the vacuum pump, an additional vacuum flask trap or a HEPA filter is recommended. Just make sure that the vacuum line is turned off when not being used.
Additionally, the filters should be replaced annually, bi-annually, or whenever there is evidence of deficiencies, including failure, filter blockage, or wetness. When replacing the filters, make sure to dispose of the old ones as regulated medical waste, radiological waste, or hazardous chemical waste.
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Empty The Flask Into The Laboratory Sink
Once the flask is two-thirds full, discontinue the use of the vacuum flask. Add a small volume of household bleach in order to achieve the final, less than or equal to 10%, bleach concentration. Leave it to sit at room temperature for at least 30 minutes to ensure that the disinfection is completed successfully. Once that’s done, empty the flask, the treated liquid waste, inside the laboratory sink. Make sure you use caution while doing that to minimize splashing. Apart from that, you should always use gloves, a lab coat, and safety glasses while working on the procedure.
When you are done with emptying the contents in the sinks, rinse the discharge thoroughly to ensure nothing is left behind on it. Furthermore, the flasks should be cleaned and emptied out weekly or whenever full to prevent overflow.
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References
blink. (2021, July 1). Biosafety: How to Disinfect Tissue Culture Media in Vacuum Flasks. blink. Retrieved October 6, 2021, from https://blink.ucsd.edu/safety/research-lab/biosafety/decontamination/flasks.html
