[Cytometry] Bacteria Detection

Nebe-Von-Caron, G g.nebe-von-caron at alere.com
Sun May 8 13:35:09 EDT 2011


Considering that I had no problem on the Aria's and LSR's I used I would
not think it is necessarily an electronic problem. Send me a set of
fcs/listmode data to look at for a more specific comment. Did you follow
the comments regarding and I'll try to figure out what happens. Not sure
if my reply at the time went to the list, but do you trigger on side
scatter etc? http://onlinelibrary.wiley.com/doi/10.1002/cyto.a.20696/pdf

Regards

Gerhard Nebe-von-Caron


-----Original Message-----
From: cytometry-bounces at lists.purdue.edu
[mailto:cytometry-bounces at lists.purdue.edu] On Behalf Of Mary K Stewart
Sent: 06 May 2011 17:43
To: cytometry at lists.purdue.edu
Subject: [Cytometry] Bacteria Detection

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.

Cheers,
Mary Stewart, Graduate Student
Cookson Lab
University of Washington

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