[Cytometry] Bacteria Detection

Mary K Stewart mks2 at u.washington.edu
Fri May 6 12:42:45 EDT 2011

This is regarding a post from April; I have had the same problem detecting the negative sub-population when analyzing heterogeneous bacterial populations that include some very bright cells.  It definitely seems to be specific to the digital cytometers.  I have found that the biexponential plot function in FACSDiva allows me to visualize the full range of fluorescence with my cells.  I make PDFs of the plots for presentation purposes, as FlowJo doesn't have the same capability and I don't have Diva available to me when I'm analyzing my data.  You can achieve sort of the same thing by altering the width basis in your FlowJo plots, but it doesn't look nearly as nice.  I have asked several folks at BD why this happens (all our cytometers are BD instruments) and it seems to be a mystery.  I am using only fluorescent proteins, no staining at all, so it isn't an issue of residual antibody aggregates.

Mary Stewart, Graduate Student
Cookson Lab
University of Washington

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