I started working with BV2 and I'm having huge contaminations problems. excluding sources of contaminations, although I confirmed that the medium was not contaminated, nevertheless I always filter the culture medium, sterilise by UV to all the materials and clean with ethanol, however, even after doing these things I keep having contamination few days after splitting the cells. sometimes few wells, other times full plates are contaminated.
I'm running out of ideas.
Can anybody help?Please?
thank you Saber for answering. I've consider that they were contaminated before, fortunately is not. But I'm inclined to the incubator. Thanks for your insights
Check to see if your incubator has a HEPA filter and replace it if it does. You should also be sure to replace the vent filter and the sample port filter if equipped (both often overlooked). Also, have the hood checked for particles. Often when a hood is certified, it is certified for proper air flow but it may not be tested for particle penetration of the filter. The water pan can also be a source of contamination long before there is visible growth. The lab air can also be a problem. When you transfer your plates between the hood and incubator its quite easy to get contamination if well plates if the lab air is too dirty. This won't occur in t-flasks since they have a dry membrane filter on the cap but the well plate lid has to stand up a bit from the well plate base to allow gas exchange which makes it more vulnerable to contamination when outside your 'aseptic' spaces. We installed our own filters in the lab ceiling diffusers because of the unpredictable condition of the air in our old building. We use the Comfort First filters and like them very much.
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