Re: Flow Cytometric Detection and Sorting of Bacteria

From: Stefan Andreatta (stefan.andreatta@uibk.ac.at)
Date: Fri Jun 11 1999 - 03:23:49 EST


Hi Ragip,

There are quite a few people around who analyse bacteria and somewhat
less, who also sort them. They use all kinds of machines and protocols.

I have been working with Flow-Cytometry and bacteria for a few months
now. The instrument is a MoFlo (Cytomation, Colorado) equipped with two
water cooled argon lasers and high speed sorting capability. The stains
I have used so far are mostly DAPI (requires UV excitation) and SYTO13
(Molecular Probes). 

A crucial point may be to use fluorescence as a trigger signal instead
of side scatter (which is low) or even forward scatter (which is almost
non-existent in my current setup).

Although or bacs from natural aquatic samples are very small (frequently
down to 0.2micron) I haven't found any detection limit so far. Analyses
were compared to counts done by fluorescence microscopy. Furthermore, we
checked sorted bacteria in the microscope. To my impression, the
flow-cytometer detects even smaller cells than you would see in the
microscope, but that has not been thouroughly checked yet.

good luck
Stefan

-- 
Stefan Andreatta
University of Innsbruck, Institute of Zoology and Limnology
Technikerstrasse 25, A-6020 Innsbruck, Austria
phone:++43-512-507-6122; fax:++43-512-507-2930
http://zoology.uibk.ac.at/limno/

-------------------------------------------------------------------------------
Ragip Unal wrote:
> 
> Does anyone know any flow cytometric metod which can detect and sort
> bacteria ? If there is waht is the detection limit of detecting bacteria ?
> All your will be greatly appreciated.
> 
> Ragip Unal
> Food Microbiology and Safety Laboratory
> The Ohio State University
> Department of Food Science and Technology
> 122 Vivian Hall, 2121 Fyffe Rd.
> Columbus, OH 43210



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