bacteria by flow

Larry Arnold lwarma at med.unc.edu
Tue Nov 15 15:43:13 EST 2005


We routinely look at bacteria on our two MoFlos.  We have looked at 
E. coli, Pseudomonas (plant pathogen), Erwinia, and Borrelia (Lyme 
disease agent).  We use FSC and SSC log and trigger on the 
SSC.  Bacteria are well resolved from noise (about 1.5 
decades).  Investigators have used GFP bacteria in expression cloning 
strategies which have worked very well (see Chang et al. PNAS 
102:2549) as well as surface labelling with Alexa488, fluorescein and 
PE antibodies.	We have never found it necessary to trigger on 
fluorescence to find the bacteria.  For good FSC signals in a 
jet-in-air sorter it is necessary to have the tip as close to the 
laser intercept as possible and keep the drop drive amplitude as low 
as possible to prevent laser light scatter from the stream waves 
getting into the FSC detector.	In general we have found it quite 
easy to sort bacteria with our systems.

Regards,

Larry

Larry W. Arnold, Ph.D.
Research Professor and Director, Flow Cytometry Facility
Department of Microbiology and Immunology
CB# 7290
University of North Carolina
Chapel Hill, NC 27599
Phone: 919-966-1530
FAX: 919-962-8103 





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