E. Coli. on Elite

Rochelle A. Diamond diamond at its.caltech.edu
Mon Aug 3 10:14:17 EST 1998

Good morning Flow-ers,
I need a little help.  Our facility has only analyzed and sorted mammalian
cells and yeast in the past.  We now have two investigators that need to
analyze and sort E. Coli and other bacteria (waste water type).  

On our first attempt, I was unable to distinguish the bacteria from laser
noise or sheath noise, having to crank up the forward scatter gain and
voltage to extreme values.

Our next attempt, I first located 1 micron beads on the scatter.  The CV's
are very large at the settings that I used to see them.  Then we put on
the bacteria and I could not distinguish them away from buffer alone.  

What do people do for this?  Do I need different forward scatter masking?
A different forward scatter detection system.  Smaller beads to set up on?

Any suggestions would be useful.  

Thanks in advance,
Rochelle Diamond
Caltech Biology 
Flow Cytometry/Cell Sorting Facility

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