Hello all,
I have been seeing small dark dots scattered all over my fibroblast cells. The growth rate appears normal, there is no clouding, but the pH appears to drop within 24 hours. I stained for DNA with PicoGreen (since we don't have DAPI at the moment) and found tiny dots all around. I am attaching one of these images. Can anyone verify if this is a mycoplasma contamination, or if not, what it could possibly be? Thanks!
This could very well be a mycoplasma infection with one of the cell-adhering species such as M. hyorhinis. As I see the Picture, the dots are lining the cell membrane, typical for mycoplasma infection seen in DAPI or Hoechst staining. Have no experience with PicoGreen, however.
The rapid decrease in pH would be consistent with an M. hyorhinis infection as this species is a glucose fermenter.
Best thing you can do is to find a lab that can test your cells with a PCR test - do not rely on the enzymatic or immunological tests on the market.
Good luck
This looks potentially like Mycoplasma contamination, and I concur with Dr. Jensen that the rapid acidification is further suggestive. The best confirmation would be mollicutes-specific 16S or RpoB PCR. Sorry to add to the suspicion!
Thanks for the query.
There is only way out that you must go for mycoplasma group specific PCR test. The protocol of how to conduct it is available in the references. It is with 16S rRNA. The product size probably of 270 bp will confirm the contamination of cell line.
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