I'm differentiating my cells which require temperature induction of 38 degrees for 14 days. I'm seeing weird stuff in my images. I hope the big cells are my differentiating podocytes which look different from its undifferentiated cobblestone shape.
but then i see these black dots in between cells. i have asked senior members in my lab what are these and one of them opine that it is cell debris (which is the case as lots of cells are dying at 38 degrees).
any opinions? the black dots do not 'move'. 24 hrs after changing media, media is still pinkish RPMI but less 'red'
Well if you had a yeast infection you may say spherical bodies growing in connected chains (3-6 or more) and bacteria normally looked rod shape. In either case you should be able to visualize them under a well aligned light microscope. You can try staining with dapi or do some electron microscopy to cofirm. I refer you to this question posted prior.
https://www.researchgate.net/post/Can_somebody_help_me_to_identify_the_type_of_contamination_from_a_cell_line_in_my_lab_Is_it_bacterial_yeast_mycoplasma
My cells went from looking like the below attached undifferentiated morphology to the cells posted at the top question section. Today all the cells died. Initially when i see that the cells were "elongating", i thought the cells were differentiating. But now with so many cell death occuring i suspect its some sort of viral infection or mycoplasma.
I will restarting my differentiation program. But any further advice is appreciaed
A quick test would be to stain with DAPI or Hoescht 33342 to see if these same small black dots are positive for nucleic acid. If positive, I would screen these further for mycoplasma. Our group had similar issues a couple of years ago.
most probably mycoplasma contamination!
For, yeast infection looks spherical bodies growing in connected chains and bacteria normally appeared rod shape with rapid movement. You can see both under high magnification.
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