Re: Hoechst toxicity
Howard Shapiro (hms@shapirolab.com)
Thu, 17 Oct 1996 19:32:40 -0400 (EDT)
The story on Hoechst toxicity goes back a long way; I believe it was in the
late 1970's that the Los Alamos group noted that viability of sorted vitally
(Hoechst 33342-)stained cells was compromised when laser power above 100 mW
was used. Augmentation of the mutagenic effects of acridine orange by blue
light (and of methylene blue by red light!) was described in the 1940's and
'50's.
In principle, one ought to be able to minimize damage to sperm by use of a
flow sorter with efficient optics; the best resolution of sperm stained with
Hoechst dyes and DAPI is generally obtained with the Partec and other arc
source instruments. However, the fluidic sorters available with more
optically efficient flow cytometers won't run as fast as conventional
droplet sorters, and certainly won't compare with high speed sorters.
Sulfhydryl compounds and other free radical trapping agents might lower
toxicity, but, if it remains a serious and pressing problem, you might want
to have some of the abundant local talent build you an optically and
fluidically efficient sorter.
-Howard
CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
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