Colleagues, Recently we purchased a new FACSCalibur which runs Cellquest on a 400MHz G3. I noticed that during initial acquisition of a sample there is a 'spurt' of activity that produces an image of low fsc vs. ssc on such a plot which within several seconds equilibrates to a normal appearance of midscale when acquiring normal mouse PBL's. BD tells us and I agree that this is simply a result of the machine equilibrating pressure and that most wait a few seconds before acquiring after placing the sample on the intake port. In fact this also relates to the question that has appeared elsewhere on this list concerning carryover from sample to sample. It is clear that this fact was not apparent on older machines because the equilibration had resolved itself before the slower processors could pick it up. Thus my question is, was it necessary to wait a few seconds on the older machines anyway because the finalization of the 'data-values' was commited to by the cytometer or wasn't it necessary because only now with the more rapid CPU does it become truly necessary to wait a few seconds for equilibration prior to acquisition? Gene/UCONN Health
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