We recently acquired a BD LSR. Switching over from a Facscan is a bit like patting your head and rubbing your stomach- in the opposite directions. The visual cues are a bit off from the usual Fascan display, and I'm having trouble getting my bearings. Running your standard lysed whole blood prep, I find it practically impossible to achieve anything close to the "Facscan traditional" 3-part Dif display. At the most I can get Lymphs & monos, but no grans. If I manage to have all three in the view- then the monos overlap the lymphs and cannot be gated separately. More disturbing (to me) is that I cannot get the lymphs off the SSC low axis. I've pumped up the PMT voltage, then tried toggling throught the 2-4-8 log scale. All I've managed to so was increase the spread between the populations, or smear out the lymphs- but the population itself will NOT move up the axis. What am I doing wrong? I'm also having a heck of a time comping the APC from the FL3 channel (using FlowJo). All the other parameters look fine, this one won't budge. If anyone has any special tips-please pass them on! Bunny Cotleur Cleveland Clinic Foundation Neurosciences NC30 9500 Euclid Avenue Cleveland, OH 44195 (216) 444-1164 cotleua@ccf.org
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