From: Howard Shapiro (hms@shapirolab.com)
Date: Tue May 14 1996 - 19:49:38 EST
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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