We use PI at final concentration 2ug/mL in FL2 detector. Our negative population is <30 fluorescent units (log scale) and have not observed any shift, perhaps this shift observation is a compensation problem. We generally analyze PI on a two-axis dot-plot (PI vs SSC) and do observe a spectrum of fluorescent intensity which morphologically we have correlated with cellular damage. Mid-range PI fluorescence, 2-3 log, cells have 'ruffled' membrances, swelling, and becoming vacuolated. The extremely bright, 3-4 log, are predominately cellular debris. We are also using 7AAD (ViaProbe, PharMingen) and have found good correlation with PI. April Translational Research & Flow Cytometry Blood & Marrow Transplantation Univ Texas MD Anderson Cancer Center
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