Flow will give you the most accurate count possible due to it's numeric
resolution. Whether what you count is what you think it is that is a different
matter. There are a number of dye combinations one can use to get counts of
subpopulations with different functional states. Your success to differentiate
intact and permeabilised bacteria with the kit described depends on the ratio
in which you use the dyes and the emission filters of fluorescence channels you
use for their analysis. To comment on your case you would need to get in touch
for a few more details on your situation.
For a fairly comprehensive resource on this matter you might want to download
for free the special issue from the journal of microbiological methods on
bacterial analysis on the single cell level from
http://www.elsevier.com/locate/jmicmeth
Regards
Gerhard Nebe-von-Caron
Research Scientist
Applied Science & Technology Group
SEAC - Safety and Environmental Assurance Centre
Unilever Research, Colworth Laboratory, Sharnbrook, UK - MK44 1LQ
Tel: +44 (0)1234 264822 Fax: +44 (0)1234 222552
E-mail: Gerhard.Nebe-von-Caron@unilever.com
-----Original Message-----
From: Amy Nield [SMTP:anield@cpd2.usu.edu]
Sent: Monday, September 17, 2001 6:40 PM
To: Cytometry Mailing List
Subject: Bacterial Counts on Flow
Hi Flowers,
I have a question that I hope I can get some help with. We have a client
that would like to use the Flow to get an accurate count on his bacterial
populations. He has a kit that he got through Molecular Probes, that stains
DNA based upon if the cell membrane is intact or not. Basically he would like
to count the live cells vs. the dead cells. When we attempted the kit we could
not get any separation of live vs. dead cells. Any help would be greatly
appreciated. If anyone knows of this kit (BacLight Bacterial Viability Kits)
and how it works please let me know.
Thank you for your help,
Amy Nield
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