Adrian Smith, We've had success with Calcofluor white (CFW) as a UV-excited live-dead dye. If you just compare %-pos with PI, CFW yields higher %-pos. On closer examination, you will see that CFW also appears to label "cell fragments" that PI does not (because these cell fragments possibly do not contain DNA). One reference from 1994 lists these sources: American Cyanamid, Difco, Polysciences, Remel, and Sigma (http://www.nejm.org/content/1994/0331/0021/1459a.asp). Interestingly, this reference also lists another source for CFW, "... Liquid Tide with Bleach Alternative, but any product containing a nonchlorine 'optical bleach' should do as well." For more information, consult Howard Shapiro's "Practical Flow Cytometry". He also provides a more useful reference for using CFW with animal cells. Eric >Hi all, > We are currently evaluating a B-D LSR in order to decide >whether it meets our requirements. One possible configuration we are >considering would use DAPI excited by the UV laser (325nm) for >live-dead discrimination in place of PI. I know we don't need to >devote a channel to PI but some users would prefer to have a separate >live/dead channel and in most experiment we wouldn't be using the UV >laser for anything else. > >My question is has anyone used DAPI for live-dead discrimination? How >does it compare to PI? Are there any particular issues we need to be >aware of? > > >Thanks, > >Adrian Smith /\/\/\_ Eric Van Buren, aa9080@wayne.edu \ \ \ Karmanos Cancer Institute and Immunology & Microbiology \_^_/ Wayne State University, Detroit, Michigan, USA
This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:01:07 EST