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@ski.mskcc.org
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