Usually log side scatter gives more meaningful clusters Log forward scatter is only noise in most cases. The best method to check if your signal is real this is to double label with a DNA stain and trigger on DNA to see the different light scatter clusters of DNA pos. and neg events. In order to understand the limits of your instrument try to run sterile filtered sheath to see when your side scatter threshold starts delivering signal. Then clamp the sheath fluid, ideally stop it altogether. If your flowrate goes down you have signals coming from your sheath and your system needs cleaning. Regards gerhard nebe-von-caron -----Original Message----- From: Moss, Delynn M. [SMTP:dmm3@cdc.gov] Sent: Friday, July 20, 2001 1:42 PM To: Cytometry Mailing List Subject: Microsporidia Hi, I am interested in a response from anyone who has flow experience on bacteria or microsporidia that measure one micron in size. We have successfully performed flow (FACScan) on microsporidia that measure two microns, but this was close to the minimum size limit on the FSC profiles when using LOG mode. The purified populations become more dispersed when LINEAR mode is used on the scatter profiles of which we prefer not to use. Is it better to use LINEAR or LOG mode for scatter on one micron organisms using a FACscan? Thanks in advance. Delynn Moss Centers for Disease Control Division of Parasitic Diseases
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