Chris You are setup correctly. However, you need to plot GFP vs autofluorescence (e.g. off the PE channel) to separate the high auto cells from the low GFP+ cells (thee would be overlapping in a one parameter histogram). Autofluorescent cells will be on a 45 degree line out from the "negatives" while the GFP population will be shifted off the 45 in the GFP direction (if GFP is on the X-axis then GFP+ will be shifted to the right off the 45). You may also apply some compensation (PE-%GFP) to further separate. Larry At 05:34 AM 12/15/2006, you wrote: >I was wondering if anyone might have advice on filter selection to >improve the discrimination of weak GFP expression and >autofluorescence. We're using an EPICS XL with a 488 laser on >mammalian cell lines expressing emerald GFP (peak excitation 487 nm, >peak emission 509 nm). > >The filters we're currently using for this are a 550 dichroic >longpass a 525/15 bandpass. Is there room here for improvement? > >Thanks, >Chris > >........................................... >Christopher Logg >University of California, Los Angeles Larry W. Arnold, Ph.D. Research Professor and Director, Flow Cytometry Facility Department of Microbiology and Immunology CB# 7290 University of North Carolina Chapel Hill, NC 27599 Phone: 919-966-1530 FAX: 919-962-8103Received on Mon Dec 18 14:38:00 2006
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