Greetings. I would like to find out what the consensus is on the loss of fluorescence of post sorted viable cells during re-analysis. I have seen this "quenching or bleaching" several times on viable cells labeled with FITC, PE, or PE-CY5 tandem dyes. Is it normal to adjust the pre-sorted region (within a few channels) to check the purity of the sort OR does one leave the region in the exact position from the original sort equation? The laser used is a 15mW air-cooled argon, pressure is 15psi, Na azide free sheath fluid, all viable cells are maintained at 4oC, "normal" surface immunophenotyping with directly labeled MAb is employed . Any and all comments are appreciated. Thank you in advance. O. Joseph Trask, Jr. The Ohio State University Analytical Cytometry Laboratory 416 Comprehensive Cancer Center 410 West 12th Avenue Columbus, Ohio 43210 Tel: 614/292-FLOW(3569) Fax: 614/292-7335 E-Mail: cytometry@osu.edu
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