Question from a novice: There are several papers which describe ways in which cell concentration can be determined using FCM. When quantifying cells, one must identify "events" which are assumed to be the cells of interest. How does one validate, quantitatively, that the events are in fact the cells of interest. For example, if we count 1x10E6 bacteria, how do we know that there aren't 9x10E5 bacteria and 1x10E5 small fungi that scatter and fluoresce in a similar fashion? My assumption is that one would need to program the cytometer to sort X events believed to be cells of interest. We would then need to quantify the cells using fluorescence microscopy. Any suggestions?
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