Chlamydia antibodies

Gerhard Nebe-von-Caron Gerhard.Nebe-von-Caron at Unilever.com
Thu Mar 21 13:23:47 EST 2002


I actually use Chlamydia as a nice example to demonstrate the advantages for
multichannel triggering in the microbiology tutorial. I use the SYVA Microtrak
kit which it is unfortunately FITC labelled but sufficient to count elementary
bodies by flow cytometry. If you want to use a green nucleic acid stain you
might want to add a second step antibody of your choice.

Gerhard Nebe-von-Caron

Research Scientist
Applied Science & Technology Group
SEAC - Safety and Environmental Assurance Centre
Unilever Colworth, Sharnbrook, Bedfordshire,  UK - MK44 1LQ

Tel: +44 (0)1234 264822, Fax: +44 (0)1234 222552
E- mailto:Gerhard.Nebe-von-Caron at unilever.com



-----Original Message-----
From:	Chance, Tim [SMTP:jtc3 at CDC.GOV]
Sent:	Tuesday, March 19, 2002 7:17 PM
To:	Cytometry Mailing List
Subject:	RE: Chlamydia antibodies


Does anyone know of a source for flurochrome labeled MoAb to Chlamydia
bacteria. Preferable PI or PercP tagged.
Tim Chance
CDC

-----Original Message-----
From: Howard Shapiro [mailto:hms at shapirolab.com]
Sent: Thursday, February 07, 2002 7:38 PM
To: cyto-inbox
Subject: Re: Red He-Ne laser vs. red diode



Ana Salas asked-

>Which source of red
>light is nowadays more suitable for flow cytometers in terms of power,
>stability (noise), life, mainteinance and prize?
>Facscalibur has a red diode 635nm but I think that new LSR is provided
>with an He-Ne laser 633nm.

The shortest answer is that whichever laser the manufacturer will sell you
with some reasonable warranty should do the job.

He-Ne lasers are larger, consume more power, and usually cost more per
milliwatt than red diodes; they have nicer beam shapes (TEM00), and they
don't have much (but do have some) long wavelength incoherent emission at
wavelengths in the region of some of the fluorescence you're trying to
excite with the primary beam.  Noise on air-cooled He-Ne's with reasonable
power is about 1% RMS.

Diodes, while very small, more energy-efficient, and less expensive than
He-Ne's, have ugly beams, which can be made reasonably smooth with
appropriate optics, and can be made very quiet (a few hundredths of one per
cent RMS noise), but they do emit long wavelength LED glow which usually
requires that they be used with band pass excitation filters, and they can
become unstable due to mode hopping.  Diodes also vary over a range of a
few nanometers in emission wavelength (635-640 nm); He-Ne's are really 633
nm, period.

Both He-Ne and diode lasers should be good for over 10,000 hours of
operation, but there seems to have been a higher failure rate among diodes,
at least until recently.

In general, the user isn't the one who puts the red laser into her or his
instrument; the cytometer manufacturers do that, and they deal with the
laser system manufacturers to get the specs they need.	The FACSCalibur has
extremely good red fluorescence sensitivity using a diode, and, if I'm not
mistaken, it is a diode that is the standard red excitation source in the
LSR, which also uses a He-Cd laser (*not* He-Ne) for UV - but if B-D is
putting a red He-Ne into the LSR instead of the diode - possibly for more
power - it should work just fine.

-Howard


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