Julie It looks like you still have the Fl-3 channel free to use. We add 7AAD to both exclude dead cells, but also allowing the Fl-3 channel to be a dump for autofluorescent cells (Fl-3 vs FL-1). Gating on low FL-3 populations will give you both viable cells and non-autofluorescent cells. It works for us ! Andy >Hey ya'll, > >We have mouse peritoneal exudate cells that we are trying to stain with >Mac3 PE, MHCII PE, CD45 APC and CD3 FITC (not all at the same time). We >get a huge autofluorescent population (unstained cells) straight out of >the mouse that shows up on FL1 vs. FL2 histogram. We cannot compensate >this out. Our voltages are reasonable (between 450 and 550). I will take >any advice. > >Thanks, > >Julie > >Julie G. Nelson >Research Coordinator >Flow Cytometry Facility >Center for Tropical and Emerging Global Diseases >University of Georgia
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