A potential cross-contaminated infection with no obvious reason. The cells that I am using is a normal human fibroblasts. I scanned the control sample after fixation, staining with gamma-h2ax and then DAPI. Small spots were stained as well around the nuclei. Is this considered as a mycoplasma infection?
There are many kits available in the market to detect mycoplasma. I would suggest you to use them and then confirm.
If the cells are already on a coverslip, then the extracellular DAPI staining is a very strong indication of an infection. There are many kits available from many companies one of which is Sigma.You can check your alive cells taking an aliquot of the culture media and doing PCR with one of the kit of your choiche.If you want to have a not expensive and quick answer to you problem you can grow some fresh cells on coverslip and then stain them with Hoechest or DAPI. If you still see the cytoplasmic speck florescence may be you can consider buying one of the kits (they are actually expensive). But on cells already fixed you can't perform the assay and to waste ab or other reagents in continuing your experiment will be only a waste of money and time because you will get unreliable data
Hi guys,
Thanks very much. Some of my cells fixed on small dishes covered with coverslip.s In some of them, I found a spready blue dapi spots around the nuclei. I have not checked the live cells yet.! We sterilised most of everything in the lab and destroy all unsealed tubes, media and incubated cells. I am running some experiments thus still keeping some of my cells. I hope this infection is not affecting my results!
Thanks.
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