Due to the intracellular growth of those buggers and the biohazard involved I would not consider live staining. Also the usually transmitted elementary bodies do not label up with an esterase substrates like ccfse. Whilst I haven't measured their phagocytosis I have measured them in their host cells using the SYVA kit (now Behring diagnostics) containing an FITC labelled antibody for the major outer membrane protein (MOMP). It does not allow you to exactly quantify the number of intracellular bodies but you can distinguish between positives and negatives. The enumeration of the bodies is difficult as they usually escape light scatter detection, but if you have a sorter and can tweak your sample flow volume you can detect them by triggering on the green fluorescence of the antibody labelled cells. Pictures on analyzers tend to be more messy. Regards Gerhard -----Original Message----- From: Andreas Thiel [SMTP:thiel@drfz.de] Sent: Monday, September 18, 2000 10:20 AM To: Cytometry Mailing List Subject: labelling of Chlamydia bacteria Hi flowers, does anyone have experience in permanent fluorescent labelling of live bacteria, in particular Chlamydia trachomatis ? We would like to quantify infection rates in monocytes. Any help would greatly be appreciated. Thanks in advance! Andreas Thiel Dr. Andreas Thiel Clinical Immunology Schumannstr. 21/22 10117 Berlin Germany phone ++49 30 28460 682 email:thiel@drfz.de
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