Re: DAPI as a viability exclusion dye

From: Howard Shapiro (hms@shapirolab.com)
Date: Thu Mar 15 2001 - 17:06:05 EST


Joseph Webster and Mark Kukuruga asked about the viability of cells stained
with Hoechst 33342 for DNA content and sorted after measurement in a UV
illuminating beam.  There was an old publication from Los Alamos which, if
I remember correctly, concluded that CHO cell viability was decreased
substantially at laser powers above 100 mW; this presumably involves
interaction of the UV light with the DNA-bound dye.  There is much older
literature showing that acridine-stained cells can be killed by blue light,
and that cells stained with methylene blue (which does bind to DNA) can be
killed with red light.  The same cells are not harmed substantially by
brief exposure to the dyes if they are not also exposed to light.

Since DAPI does not normally get into viable eukaryotic cells, there should
not be much reason to worry about effects on the nonstaining viable cells
when DAPI is used as a "nonviability" indicator.

-Howard



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