right FCM for natural bacteria

From: Stefan Andreatta (Stefan.Andreatta@uibk.ac.at)
Date: Thu Nov 20 1997 - 04:20:40 EST


Dear listmembers,

We are going to purchase a flow cytometer with sorting capabilities 
for use in aquatic microbial ecology. Our main task is to measure 
bacteria from natural samples (DAPI, SYTO13, CY3,...). 

Unfortunately these organisms are very small (down to 0.2 microns) 
and they do not live in the water all by themselves - there will 
be lots of algae, flagellates, detritus etc.

So we need to have good sorting capabilities to check what we are 
actually measuring in the microscope, but the big problem (I feel) 
is the instrument's sensitivity.

I would like to ask experienced users out there:

1- Which features, do you think, are of particular importance to our 
application. (PMT for forward scatter is certainly one). Other 
important features?

2- What do you think of jet in air vs. cuvette sensing. Flat surfaces 
should enhance scatter measurement, but seemingly only Coulter is 
ready to provide a sorting(!) cuvette. Other important features?

3- We need three excitation wavelengths (UV, 488, 514 or higher) for 
various experiments, not all three at the same time but all possible 
combinations of two of them. This will involve a lot of laser 
adjustment. How complicated is it on different instruments e.g. to 
shift a laser beam from second to first sensing position?

4- I would highly appreciate hearing about any relevant experience on 
different instruments. What we have in mind is to bye one of the 
following (in alphabetic order): 
-Becton Dickinson FACS Vantage
-Coulter Epics Elite ESP
-Cytomation MoFlo MLS.

Thanks for any hints
Stefan





----------------------------------------------------
Stefan Andreatta
University of Innsbruck, Austria
Institute of Zoology and Limnology
Technikerstrasse 25, A-6020 Innsbruck
phone: +43-(0)512-507-6122
fax:   +43-(0)512-507-2930



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