Re: viability on fixed cells

From: Julie Oughton (oughtonj@ucs.orst.edu)
Date: Thu Mar 16 2000 - 13:46:55 EST


Yes--it is possible! I had excellent results using 7-AAD to stain dead
cells prior to fixation.
The leakage of 7-AAD to live cells can be blocked by using non-fluorescent
actinomycin D in the media. I followed the technique outlined by Schmid et
al. (1999) in Cytometry 35:64-74.

Julie Oughton
Oregon State University


>
>
>
>At a recent flow cytometry workshop in the Research Triangle Park area  the
>following question was asked:  Is it possible to measure viability in a cell
>population and then fix the cells prior to analysis by flow cytometry.
Can the
>dye stay in the dead cells without leaking into the live cells?  I
remember that
>Dr. Allan Pollock worked on these techniques in the 1980's . Does  any one
have
>any knowledge or information on this type of protocol. The research groupe
wants
>to apply this technique on AIDS blood. Thanks
>Bob
>
>Robert M. Zucker, Ph.D
>U.S. Environmental Protection Agency
>National Health and Environmental Effects Research Laboratory
>2525 NC Highway 54,  MD 72
>Research Triangle Park, North Carolina, 27709
>tel: 919-541-1585 fax 919-541-4017
>e-mail: zucker.robert@epa.gov
>
>
>



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