Marty, For my first attempts at sorting gfp cells I used 500mW, and reanalysis of the two fractions looked like we hadn't sorted at all. I tried again w/ 50mW (probably unstable on a 90-5) and it worked beautifully, >90% purity. I blame photobleaching. If you're already using comparable laser power then I'm at a loss! Good luck, Steve > > One of our users who screens a lot of his cell lines using GFP and cFP > finds that upon re-analysis of his sorted population, most, if not all, are > at a lower intensity, many times even below the sort window. Having > verified our sorting conditions we are wondering if photo-bleaching of the > fluorochrome is responsable for this. I have not checked the literature on > this, and would appreciate hearing about any experiences or getting > references. > > Marty Bigos > Beckman/Stanford Shared FACS Facility > +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Steve G. Hilliard (706) 542-9474 University of Georgia Cell Analysis Facility flowman@uga.edu URL: http://floweb.cb.uga.edu/Floweb/ +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
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