There has actually been an article in cytometry showing that it can be done
with a jet in air system, so with some restrictions.
Regards
Gerhard
See
Flow cytometry of bacterial cells: Comparison between different flow cytometers
and different DNA stains
Bernander R, Stokke T, Boye E
CYTOMETRY
31: (1) 29-36 JAN 1 1998
Abstract:
The DNA content and Light scatter from individual Escherichia coli cells were
measured in two flow cytometers with different configurations, The DNA content
could be measured with similar resolution either in an Argus now cytometer
equipped with a mercury lamp, or in a FACStar flow cytometer with two argon
lasers as
light sources. In contrast, Light scatter measurements appeared to be a good
measure of cell mass only in the Argus instrument.
Three DNA stains were compared:, DAPI, Hoechst 33258, and mithramycin A
together with ethidium bromide, All three stains yielded DNA histograms of
similar
quality in both flow cytometers. Optimal results required that the stain and
cell concentrations were kept similar, that a fixed rate of sample introduction
was used,
and that a period of equilibration was allowed during running of each sample.
The results demonstrate that conventional, laser-based flow cytometers may be
used for high-resolution measurements of bacterial DNA content, thereby making
flow cytometry available to an increased number of research groups working with
prokaryotes. (C) 1998 Wiley-Liss, Inc.
-----Original Message-----
From: Louis King [SMTP:kingl@pilot.msu.edu]
Sent: Friday, June 01, 2001 9:23 PM
To: Cytometry Mailing List
Subject: Need help on quantitative bacterial DNA measurements
To: cyto-inbox
I have a request from a person for flow cytometric help in identifying the
number of chromosomes replicating DNA in a bacterium. He has altered
proteins he/they think are involved in bacterial chromosome replication so
they want to compare the parent strain to the altered strain. He wants to
be able to establish significantly altered chromsomal replication patters
having 3 or more replication forks.
Any body out there in the US doing this? How about world wide?
Has anybody out there tried to do this on a jet in air FACS unit? (You may
laugh now. If by some miracle somebody did get even close to making this
work--drop me a line will you?)
Does anybody have a microscope based FACS unit that will do this? Partec
or other variety?
Any useful comment from experience will be greatly appreciated.
Louis King
Louis King, Ph.D.
Michigan State University Flow Cytometry Center
Department of Biochemistry and Molecular Biology, Rm 419
Michigan State University
East Lansing, MI 48824
Phone: 517-355-1536
FAX: 517-353-9334
E-Mail: kingL@pilot.msu.edu
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