From: Keith Bahjat (kbahjat@liai.org)
Date: Wed Dec 17 2003 - 17:35:16 EST
James, Molecular Probes has kits that can be used for this purpose. I purchased their bacterial enumeration kit, then added PI so I could distinguish live from dead bugs. You can buy their Live/Dead kit and combine this with the bacterial enumeration kit if you donšt feel like making up reagents. Since the bacterial dye is metabolically converted, youšll find that after a little practice, you can distinguish live from dead bugs without the additional PI. I used this technique for quantifying CFUs of Listeria monocytogenes stocks during large batch production using our FACSCalibur. Results matched very well during parallel tests with dilutional plating. Just remember not to run things too concentrated (below 3000 per second) to avoid overwhelming the electronics. You can find more info about the kit via this link: http://www.probes.com/media/pis/mp07277.pdf Cheers Kb -- Keith Bahjat, Ph.D. Division of Cellular Immunology La Jolla Institute for Allergy & Immunology San Diego, California kbahjat@liai.org On 12/17/03 7:50 AM, "James Kirby" <jekirby@bidmc.harvard.edu> wrote: > Dear All, > I was interested in finding protocols for doing flow on bacteria to > enumerate bacterial numbers and determine viability. We have a BD > FACSCalibur. Is anyone using this particular cytometer for > bacterial analysis. What general settings are used for detecting > bacteria in assays? > > Thanks, > James
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