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Cell Fixation for DNA flow cytometry in fibroblasts - (Sep/02/2012 )

I was wondering if someone could help me with my problem.
I've been trying to read my fibroblasts from adventitia in an hemocytometer. I dont know why but my cells dont get any difference. I believe that they destroy when I fixate them with ethanol 70% as every protocol indicates. I use IP as a dyer. I used to starvate them for 24 hours and then stimulate them whit Serum for 16, 18, 20, 22 and 24 hours in order to snapshot different cell cycle times.I f someone has ever worked with this kind of cells and know any tip or special protocol I would be totally gratefull.


I'm not quite sure what you are asking - but yes, ethanol will kill the cells, so that a trypan blue stain will only show dead cells after fixation.

It is possible to do live cell FACS, but I don't know any thing about it.


Your topic says flow cytometry, but then you are talking about a hemocytometer? I'm confused, which do you mean?

At any rate, what do you mean you don't get any difference? Any difference in what?