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You can effectively use Ethidium monoazide. The way to do it is to label
the cells and then to fix them.
You will find a protocol in : Cytometry 12, 133 (1991) M.C. Riedy et al.
See the Medline reference below.
I guess that you can get some more details getting in touch with Molecular
Probes, since they have this reagent in their catalog (cat.: E-1374).
Molecular Probes e-mail: tech@probes.mhs.compuserve.com
phone: 503-465-8338 / fax: 503-344-6504 or 1-800-438-0228
Good luck !
Rene Olivier
-----------------
ARTICLE TITLE: Use of a photolabeling technique to identify nonviable
cells in fixed homologous or heterologous cell populations.
ARTICLE SOURCE: Cytometry (United States), 1991, 12(2) p133-9
AUTHOR(S): Riedy MC; Muirhead KA; Jensen CP; Stewart CC
AUTHOR'S ADDRESS: Roswell Park Memorial Institute, Department of Flow
Cytometry, Buffalo, New York 14263.
MAJOR SUBJECT HEADING(S): Affinity Labels [diagnostic use]; Azides
[diagnostic use]; Cell Survival; Flow Cytometry; Fluorescent Dyes
[diagnostic use]
MINOR SUBJECT HEADING(S): Antibodies, Monoclonal [diagnostic use];
=46ixatives [pharmacology]; Immunophenotyping; Lasers; Specimen Handling
INDEXING CHECK TAG(S): Human
PUBLICATION TYPE: JOURNAL ARTICLE
ABSTRACT: Flow cytometric determination of viable versus nonviable cells
in fixed samples can be accomplished by utilizing the irreversible binding
of photoactivated ethidium monoazide (EMA). EMA is a positively charged
molecule which is excluded by cells with intact membranes (viable cells),
included by cells with damaged membranes, and can be photochemically
crosslinked to nucleic acids using visible light. EMA fluorescence can be
excited using a standard argon laser operating at 488 nm and is able to be
distinguished from fluorescein and phycoerythrin. Fixation is important
when analyzing cells from a potentially infectious origin. EMA is
photochemically crosslinked and therefore unable to leak out of cells when
removed from the extracellular media, unlike propidium iodide (PI) or other
viability stains, which were heretofore commonly used. We demonstrate the
usefulness of EMA in combination with fluoresceinated and phycoerythrin
labeled monoclonal antibodies in immunophenotyping. The photoaffinity
labeling technique allows for a quick and efficient means of identifying
nonviable cells which cannot be distinguished on the basis of
light-scattering properties.
MEDLINE INDEXING DATE: 9109
ISSN: 0196-4763
LANGUAGE: English
UNIQUE NLM IDENTIFIER: 91266648
CAS REGISTRY/EC NUMBER(S):
PS: there is a problem with your e-mail address (=7F=7Fmonard@adarc.nyu.edu)=
,
you are probably using a special character that is not accepted by servers:
The original message was received at Tue, 30 Jan 1996 12:12:33 +0100
from [157.99.72.146]
----- The following addresses had delivery problems -----
<=7F=7Fmonard@adarc.nyu.edu> (unrecoverable error)
----- Transcript of session follows -----
... while talking to phri.nyu.edu.:
>>> RCPT To:<=7F=7Fmonard@adarc.nyu.edu>
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____________________________________________________________________________=
__
Rene Y. Olivier
Unite d'Oncologie Virale
Institut Pasteur
25-28 rue du Dr Roux
75724 Paris Cedex 15
=46rance
Tel: 033 1 45 68 83 75
=46ax : 033 1 40 61 34 65
Secr: 033 1 45 68 87 41
E-mail: rolivier@pasteur.fr
____________________________________________________________________________
__
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