more contamination owes - spherical cell like contamination (Oct/07/2005 )
Hi
I have been going crazy over some weird contamination in my cell culture plates.
Some spherical cell like refractile stuff keeps showing up in all my plates even after reapeated washing. It is too big to be yeast and the media also doesn't turn cloudy. It persists even after treatment with 2.5ug/ml fungizone.
Mostly, I see them floating in the media, but sometimes a few are attached to the plates near my cells. I have used cells from the same stock before and have had no problems.
This is driving me crazy and I am totally wasting my time and unable to do any experiments. I desperately need these cell lines and cannot throw them out.
Somebody please help!!
Thanks.
this is probably not your problem, but are you QUITE certain you are not looking at air bubbles?
i had the same problem once after switching pipet brands (different sized bore) and it took me almost two weeks to figure out that it wasn't some strange sort of yeast; i tried EVERYTHING and then one day I realized I always saw the most 'contaminant' right after a media or buffer rinse or change....
Man, don't you just HATE that kind of thing? It is one of those things that will only happen to you once...
These dont sure look like air bubbles to me, but I cant figure out what they are:(
have you done all the obvious stuff, like incubate uninoculated media, and check your flasks, pipets, etc for contamination?
Sorry for the delay in replying. I have tried everything. The uninouculated media does not have any contamination. But whenever I plate any kind of cell, these stuff are still there floating around and sometimes attached to the cell surface.
I still havent found a way out.
too big to be yeast?? how big are you talking?
where did you get your cells? can you trust the source?
are they contaminated with another cell type? for example, our keratinocytes usually have a few fibroblasts floating around, but only a very tiny number. Until one day I let a culture plate go a little past senescence and tried to subculture it (back when I was young and carefree and unwise to the ways of tissue culture)...my keratinocyte growth was wimpy and the fibroblasts took over the culture plate within a couple days. I initially thought that was contamination too.
have you tested your trypsin? your PBS? all that other stuff that is not media?
Do you think, that these are dividing cells? these cells look spherical and they can be attached or floting.
I have been going crazy over some weird contamination in my cell culture plates.
Some spherical cell like refractile stuff keeps showing up in all my plates even after reapeated washing. It is too big to be yeast and the media also doesn't turn cloudy. It persists even after treatment with 2.5ug/ml fungizone.
Mostly, I see them floating in the media, but sometimes a few are attached to the plates near my cells. I have used cells from the same stock before and have had no problems.
This is driving me crazy and I am totally wasting my time and unable to do any experiments. I desperately need these cell lines and cannot throw them out.
Somebody please help!!
Thanks.
Could you please tell me what cells are you trying to grow.
Also, these spherical refractile bodies, is there anything inside them or are they empty. If there is something inside them, what do you see. What do you think their approximate size is (50-150um in diamter). If you have any photos I would love to see them. I had similar problems a while back, with spherical bodies flaoting in the medium, sometimes they were attached to the cells. Please send me some photos to the following e-mail address bscience@hotmail.co.uk, and I will reply back asap.
I look forward to hearing from you