It seems like your cells are fine. Mycoplasma will look like a nuclear foci everywhere on your slide. Moreover you cant see an intact nucleus incase of mycoplasma contamination. The other stains might be because of overloaded DAPI or background. Try DAPI without BSA. I noticed a mitotic cell at bottom. Beautiful!! All the best!

Thanks for your answer. I stopped suspecting mycoplasma contamination for now. I hope mycoplasma will stay out of our lab forever. Also when i saw that mitotic cell, it really made me feel as happy as a child.

That's nice!! But always do care for Mycoplasma.. Because it is omnipresent and very difficult to find source. So it is always better to add low level of plasmocin to keep mycoplasma under control!!

your cells are free of mycoplasma, don't worry! good luck!!

Hi. beautiful picture.

Cansu, I will do this test in neuroblastome culture (N2A). The cells are growing very slow. Is it possible do this test whitou fixation? Just permeabilization (triton 0.2%) and incubation whith DAPI? I never did this test. What you recommended me?

Mansoor, do you have a photo the cells contamination? For we can see how is in contamination state?

Thaks

Hi jafia,

It would be better if you fix the cells and do permeabilization with 0.2% triton X 100. i have attached a image with mycoplasma contamination (google source).. 

Hi All,

Can someone please comment on whether the image of cells that is attached show Mycoplasma contamination? I had fixed cells with 4% PFA for 10 min, permeablized with 0.5% Triton X-100 for 10min and stained with 0.05ug/ml DAPI for 5min.

Cells are imaged under 40X magnification.

Thanks a lot!

Hi,

Can someone please comment on whether the image of Vero cells that is attached show Mycoplasma contamination?

Hi Alina

Looks like your cell line is having slight mycoplasma contamination. To be sure please do a PCR check that will confirm