Dr. Howard Petrie h-petrie at ski.mskcc.org
Tue Feb 11 16:39:51 EST 1997

Fellow flow cytometrists: We are working on a three laser Vantage
(Enterprise for UV/488 source, with rhodamine dye tuned to ca. 605nm; dye
and UV beams are colinear).  We are having problems with DAPI (FL5, in
linear) spilling into the APC detector (FL4, log, with 660/20 filter).
According to the spectral data I have, DAPI should lose >95% of its
fluorescence by 575-580nm, but certainly by 600nm in any case.  We are
using DAPI at the lowest concentration that will give tight CVs on fixed
cells with 2n DNA content.  I can't figure out if we might have a bad batch
of DAPI, or something else.  Does anyone else have experience using this
combination?  Any ideas what gives?  Thanks for your input, Howard


Howard T. Petrie, Ph.D.
Assistant Member, Immunology Program
Memorial Sloan-Kettering Cancer Center
Box 341, 1275 York Avenue
New York NY 10021

phone (212)639-2149
fax (212)794-4019
e-mail: h-petrie at ski.mskcc.org

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