Hi Kimmo, We made a simple modification to our LSR almost a year ago, and determined that the side population profile compared favorably to the one obtained on our FACSVantage SE. We changed the height of the pinhole in front of the detector which houses a 670 LP such that it will collect fluorescence generated from the 325 nm excitation instead of the 488 nm excitation. The Hoechst 33342 blue fluorescence is collected with the 424/44 BP (=FL5 on the LSR), while the Hoechst 33342 red fluorescence is collected with the 670 LP (=FL3 on the LSR). The only caveat with this make-shift setup is that we "lose" one of the four detectors for the 488 nm laser pathway. I am certain that BD will come up with a better solution. Teresa ------------------------------- Teresa S. Hawley, B.S. Project Leader, Flow Cytometry Research and Development Hematopoiesis Department : American Red Cross Holland Laboratory 15601 Crabbs Branch Way Rockville, MD 20855 Tel: 301-738-0425 Fax: 301-738-0444 E-mail: hawleyt@usa.redcross.org <mailto:hawleyt@usa.redcross.org> -----Original Message----- From: Porkka Kimmo [mailto:Kimmo.Porkka@hus.fi] Sent: Wednesday, February 27, 2002 2:06 AM To: Cytometry Mailing List Subject: side population wtih LSR Has anyone have experience on running a side population analysis of human bone marrow with the BD LSR? I'm using the Hoecst 33342 dye and a FITC cell surface marker and following the published protocol to the letter, but still cannot see any Hoechst fluorescence on the red channel. Filter setup? Software compensation? Any help would be much appreciated. Kimmo Porkka ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Kimmo Porkka, M.D., Ph.D. Helsinki University Central Hospital Dept. of Medicine, Division of Hematology P.O. Box 340, 00029 HUS Helsinki, Finland kimmo.porkka@hus.fi +358-9-471-951029 (HUCH) +358-9-471-71890 (Biomedicum) +358-400-492-100 (mobile) +358-9-471-71897 (fax) +1-810-283-8318 (eFax)
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