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Consensus (if you can call it consensus) seems to be that DAPI may or may not
get into living cells -- depending on the type of cell (membrane
characteristics/ glycoprotein pumps) and the patience of the investigator (that
is, DAPI penetrates slowly at best). In some cell types and with some
protocols , it doesn't penetrate living cells very well and therefore it can
be used to differentiate living and dead cells. In cases where it does get
into living cells, it will stain DNA -- but will probably not give as pretty
DNA histograms as Hoechst 33342 (although some people report better success on
this than others). Hoechst 33342 seems to be, by acclamation, the dye of
choice for looking at ploidy in living cells. Questions were raised about DAPI
not leaving the cells viable after the staining even if it does penetrate the
membrane; this is obviously a question that needs to be considered (along with
mutagenesis) for any dye that enters a living cell and binds to its DNA....
For fixed or permeabilized cells, consensus is that DAPI and PI both give good
DNA histograms.
Thanks for all your help.
Alice Givan
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