Re: cell cycle analysis
Howard Shapiro (hms@shapirolab.com)
Tue, 20 Aug 1996 22:10:59 -0400 (EDT)
>I have been reading articles that state the cell lines used have been
>staged in G1 or G0 and this justified based on a PI flow cytogram. Is this
>a good/accurate way or would AO work better, or maybe staining of cyclins?
>
Were the authors claiming that PI could tell G1 from G0 (which, at least
under normal circumstances, it can't) or simply that PI could identify cells
as being in G1 or G0 but not determine which (which is true of DNA stains in
general). DNA/RNA measurement using AO (or the combination of Hoechst 33342
or 7-aminoactinomycin D with pyronin Y) would discriminate G0 from G1 cells,
as would cyclin staining. There are other ways to do this, e.g.,
measurement of DNA and protein content of isolated nuclei using PI and FITC.
The Hoechst/pyronin and 7-AAD/pyronin techniques share the advantage with
cyclin measurements of being usable with surface or cytoplasmic
immunofluorescent staining, which is almost impossible to do with AO
(although if you have 488 and red lasers, it might work).
-Howard
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