cell number assay

Oswaldo oswaldo at iats.csic.es
Tue Sep 26 08:02:58 EST 2000



Tom wrote:
> 
> Does anybody here know of an assay - preferably elisa or similar - which
> allows you to compare the number of live cells (tissue culture) between
> wells/plates, without affecting the cells?


You are probably looking for tetrazolium salts-based assays. They
measure mitochondrial dehidrogenase activity in live cells by
colorimetry in multiwell plates. These tests can be optimized so the
color measures correlates pretty well to the number of viable cells in
culture (or that is what they claim). There are several salts available,
but the most common are XTT or MTT. I particulary have used kits from
Promega and from Boehringher (now Roche diagnostics). Both are good but
the salt from Roche (WST-1) is very stable and used as a single, ready
to use solution. Very convenient although rather expensive.

You can get plenty of information on the procedures by looking at their
catalogs and web pages. Look for cell proliferation/citotoxicity
reagents.
 
BTW, these assays obviously do affect the cells, but at least don't kill
them immediately like the methods for measuring DNA synthesis. I guess
that is what you meant by "without affecting the cells".

Cheers,

Oswaldo






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