I am growing some leukemia cell lines (MV411, CEM) and suspect that I might have mycoplasma contamination. I see black dots in my cell culture, and I am unsure if it is cell debris or mycoplasma. I understand DAPI staining can be used to detect mycoplasma in cell culture, but a lot of the protocols I am finding are for adherent cell culture. I am wondering if anyone has experience detecting mycoplasma in suspension cultures using DAPI.
I don't really think you can see mycoplasma with phase contrast microscopy if that is what you are using to describes "black dots", they are tiny tiny bugs! With DAPI staining perhaps, I've had mycoplasma contaminated cells and only after discovering this did we later realize the DAPI cells had faint blue objects separate from the cell signals. But the best way to do is to use a kit, they're quick and very convenient. Choose the one from your favorite vendor, the one we used was from Lonza (https://bioscience.lonza.com/lonza_bs/TR/en/Cell-analysis/p/000000000000186472/MycoAlert-Mycoplasma-Detection-Kit-%28100-Tests%29).
Hello Mubasher Iqbal ,
As Can Kiessling says, I am not sure you can really see a mycoplasma contamination by just looking at your cells in culture, especially suspension cultures. Maybe if your cells are heavily contaminated it might appear as an abnormal number and density of "clouds" of very tiny black dots, but I have only observed it in adherent cultures.
I would also suggest a kit to detect the mycoplasma contamination, this way you will be sure of the result. Personally in our facility we perform a PCR using a kit from VenorGem, but I think the kit suggested by Can Kiessling is way easier and quicker to use if you only have 1-2 samples to test.
Good luck with your experiments !
Spin down some cells and check for Mycoplasma contamination by qPCR. It is very sensitive.
you can also use myocoalert detection kit that does not require qPCR equiment
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