Hi Flow people! First let me wish everybody Happy Holidays and keep up the great work on this newsgroup! I ran some 3 color samples (CD3 FITC/CD4 PE/CD8 PE-Cy5) on my Beckman Coulter Elite today and I saw something that didn't make sense to me. I've attached three slides of the histograms that I got. I ran the single color positives and compensated so that the mean values where correct (see Slide 1, compensation settings #1). I then ran the samples (example of one on slide 2). There where 2 populations that didn't seem right to me (looked like they where undercompensated). I increased the compensation so that it "looked right". I then ran the single color controls again using the increased compensation values (slide 3). The mean values I got seemed to be overcompensated. I apologize for being somewhat ambiguous in the descriptions above (I'm trying to at least get out of here on time today!) but the histograms in the attachment are more informative. I've done three color work before but have never come across this problem. The user says that the antibodies are the same (i.e. not different lots within an antibody throughout the samples). I realize that alot of people are on vacation already and responses to this might be sparse but any help would be greatly appreciated (Mario? Help!) Thanks in advance! Andy (:)-(:)-(:)-(:)-(:)-(:)-(:)-(:)-(:)-(:)-(:)-(:)-(:)-(:) Andy Oberyszyn, M.S. The Ohio State University Analytical Cytometry Laboratory 416 Comprehensive Cancer Center 410 West 12th Avenue Columbus, Ohio 43210 Tel: 614/292-FLOW(3569) Fax: 614/292-7335 E-Mail: cytometry@osu.edu (-):(-):(-):(-):(-):(-):(-):(-):(-):(-):(-):(-):(-):(-) "What if the Hokey Pokey is really what it's all about?!?"
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