labelling of Chlamydia bacteria

gerhard nebe-von-caron Gerhard.Nebe-von-Caron at Unilever.com
Wed Sep 20 05:14:40 EST 2000


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 at 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 at drfz.de

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