I saw small dust like particles floating in my cell culture media (DMEM) and in the cell culture flask as well. My cells are growing fine in this media. Also, I see the same particulate matter in FBS. Is this some sort of contamination or FBS is particulate?
Hello Adity.
It happens everywhere since everyone is too busy to do only the experiments. Both lamina flow and CO2 incubator require weekly cleanness and maintenance with sterile cleaning since they are making whirling all the time. The odd corners of them are accumulating dusts, particulates, evaporated, dried salts and dried dead or live microbes.
Usually they become getting bigger and heavier and darker, precipitating on the bottom of the dishes. Lids or tops of culture dishes and flasks are allowing free flights of those particulates through the fanning action in the incubator. It may not be OK.
I doubt that FBS has the floated particulates at first place, and I also suspect that milliporing would remove the floated matters in your culture conditions.
Regards.
FBS can have fibrin precipitates, they don´t usually affect cell growth. If you mix the serum you can see particles floating and they start to decant to the bottom.
Is your serum heat-inactivated? If you do it yourself, you should allow the serum to settle down at room temperature and carefully take only the supernatant serum which does not have fibrin and broken complement proteins. I also used to have particulate matter floating in my culture vessel but I eliminated it completely by filtering the media+serum in Millipore stericup vacuum filtration unit (https://www.fishersci.ca/shop/products/emd-millipore-steritop-sterile-vacuum-bottle-top-filters-8/p-105717), after which I add antibiotics and antimycotics.
Hello Adity.
It happens everywhere since everyone is too busy to do only the experiments. Both lamina flow and CO2 incubator require weekly cleanness and maintenance with sterile cleaning since they are making whirling all the time. The odd corners of them are accumulating dusts, particulates, evaporated, dried salts and dried dead or live microbes.
Usually they become getting bigger and heavier and darker, precipitating on the bottom of the dishes. Lids or tops of culture dishes and flasks are allowing free flights of those particulates through the fanning action in the incubator. It may not be OK.
I doubt that FBS has the floated particulates at first place, and I also suspect that milliporing would remove the floated matters in your culture conditions.
Regards.
Hi
These particles are debris from cell membrane and called brownian particles and their movements are also known as "Brownian motion". They are natural and must not get confused with microbial contamination. They usually are well-observed at 400X magnification under inverted microscope. If you can see these particles with naked eyes the others have implicated possible reasons as well.
Best
You should centrifuge FBS with high speed (25000rpm for 5min) and then filter before using FBS. It will solve your problem. Actually chances of geeting contamination become high so better used centrifuged and then filtered FBS
Can you share photograph if possible ?
I think, its mostly cell debris. Make similar media with Pen/Strep. See if anything grows in the media.
- Badges
- Science topic