DAPI as a viability exclusion dye
crissman at telomere.lanl.gov
Fri Mar 16 19:54:39 EST 2001
The paper that Howard suggests is: H. A. Crissman, M. H. Hofland, A.
P. Stevenson, M. E. Wilder, and R. A. Tobey, Use of DiO-C5-3 to
Improve Hoechst Uptake, Resolution of DNA Content, and Survival of
CHO Cells. Exp. Cell Res. 174: 388-396 (1988). What was surprising
was the effect of DiO-C5-3, particularly on cells that probably have
high levels of p-glycoprotein.
At 5:06 PM -0500 3/15/01, Howard Shapiro wrote:
>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.
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