Hello, I am currently sorting bacteria expressing a promoter trap fusion library to gfp on a Coulter Epics Elite. I am interested in talking to anyone who is sorting bacteria on this machine. More specifically, I am having trouble separating non-fluorescent bacterial cells from those cells which are weakly fluorescing. I am able to successfully sort bacterial cells expressing gfp strongly from non-fluorescing bacterial cells; however, I am unable to sort non-fluorescing cells from those cells weakly expressing gfp. I think I am having this problem because the weakly fluorescing cells are overlapping into the non-fluorescent region (this non-fluorescent region was determined by using a non-fluorescent control). Any help anyone could give me would be great. Thanks, Janet janet_roggy@urmc.rochester.edu
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