Re: Sorting G0 cells?
Howard Shapiro (hms@shapirolab.com)
Tue, 14 May 1996 20:49:38 -0400 (EDT)
Years ago, I had no luck in attempting to grow cells (stimulated lymphocytes
and lymphoblastoid lines) vitally stained for DNA and RNA, respectively,
with Hoechst 33342 (about 3 uM) and pyronin Y (5 uM), which did allow
differentiation of G0 and G1 phase cells. I subsequently had occasion to
look at cells stained with Hoechst and cyanine dye (DiOC6(3)) and Hoechst
and rhodamine 123, which produce similar bivariate distributions and thus
appear usable for discrimination of G0 and G1 cells. Although I have not
had occasion to sort and attempt to culture cells stained with these dye
combinations, I believe others have done so successfully. It might help to
use verapamil or trifluoperazine to inactivate the efflux pump; you could
then use lower dye concentrations. If you try any of these approaches, I'd
like to hear about the results.
-Howard
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