Info on Tutorial for Microbial Flow Cytometry

Gerhard Nebe-v.Caron Gerhard.nebe-von-caron at unilever.com
Sun Dec 21 17:04:33 EST 1997


For anyone interrested in microbial flow cytometry:

The Venue:
ISAC XIX congress 28/Feb/1998 Colorado Springs
(see http://nucleus.immunol.washington.edu/ISAC.html)
Tutorial on Microbial Flow Cytometry,

The Program:
The aim of the tutorial is to give flow cytometry users the confidence
and the technical background to tackle the measurement of bacteria. To
achieve that there will be a theoretical part and presentations of
practical applications, accompanied by protocols and reference
literature. It is envisaged to organise some practical demonstration
with regards to instrument set-up within the week.

1. Background information : (Gerhard Nebe-von-Caron)
1.1 Technical background
Setting the environment : Requirement on labware and reagents.
Setting up the instrument : Calibration standards
Signal processing: Triggering / threshold settings, Bacterial
discrimination, back-gating
Light Scatter measurements : Opportunities and limitations
Sorting bacteria : Instrument preparations for sterile sorting

1.2 Functional and differential labelling of bacteria
Bacterial enumeration: Sample handling, disaggregation and counting
methods.
The viability concept : Measurement of reproductive viability,
metabolic activity and membrane integrity.
Bacterial differentiation : Antibody staining in of =91environmental=92
samples.

2. Practical applications :
2.1 Microbial Flow Cytometry in Biotechnology (Susann Muller)
The measurements of physiological changes in biological processes and
their influence on process control will be presented. This will cover
the measurement of cell proliferation by DNA synthesis and changes in
the cell wall structure and the prediction of =93survival capacity=94 by
assessment of cell constituents such as stored energy and cell membrane
functionality.

2.2 Antibiotic susceptibility of Mycobacterium tuberculosis and Candida
albicans (Ronald Shell)
This presentation describes a method to test the antibiotic
susceptibility of clinical isolates of the above species based on
metabolic activity. Materials and sample preparation, instrument
set-up, analysis and data interpretation will be discussed.

The team:
Dr. Susann Muller, University of Leipzig, Fakultaet fuer
Biowissenschaften
Dr. R.F.Schell and S.M.Kirk, University of Wisconsin, State Laboratory
of Hygiene
Mr G.Nebe-von-Caron, Unilever Research Laboratory Colworth

Abstract for the tutorial on Microbiology:

Despite 23 years of microbial flow cytometry the technique is still not
so widely used amongst microbiologists as it could be. Thus the aim of
the tutorial is to encourage people to take microbial cytometry on board
as an additional service that a cytometry facility can offer.
The tutorial will contain three short presentations discussing
biotechnological, environmental and medical applications and address the
fundamental question of microbial viability and cell injury. This will
be followed by a technical discussion about instrument set-up and other
useful tips and tricks. It is aimed at people that have already a basic
understanding of flow cytometry. As most cytometers currently on the
market can theoretically be used for a number of microbial applications,
the exchange of technical information should allow operators to gain
confidence in microbial analysis with their particular equipment.

Please contact myself if you require any further information

With the best wishes for a peaceful Christmas
Yours sincerely

Gerhard Nebe-v.Caron
Unilever Research, Colworth Laboratory
Sharnbrook, Bedfordshire
GB - MK44 1LQ
Tel.: +44-(0)1234-222066
FAX: +44-(0)1234-222344
E.mail: gerhard.nebe-von-caron at unilever.com



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