JC-1 fixation

Patricex Petit pxpetit at cochin.inserm.fr
Fri Jun 11 04:52:48 EST 2004


Answer to

Hello !
I will like to know if JC-1 stained cells can be fix for later acquisition.
I read a lot of literature and could not find anything about this regard.
My guess it's no, but I'm not sure.  If anybody can answer this question
for me I'll appreciate it.
Thanks
Johana

Johana Melendez, MS
Research Associate
Flow Cytometry Laboratory
H. Lee Moffitt Cancer Center
12902 Magnolia Dr.
Tampa, Florida 33612-9497
Phone (813) 745-6610
Lab. phone (813) 972-8400 x. 2005
Fax (813) 903-7140
 <<Melendez, Johana.vcf>>

Dear friend,

If you wanted to keep the relation between JC-1 dye and the detected
mitochondrial membrane potential, this is not possible. because JC-1 is a
live dye and will be released as soon as the mitochondrial membrane
potential will be abolished by the fixation.
I propose the following experiment with a dye like CMXRos...
stain your cells and measure them non-fixed in two situations, where you
added nothing and the other one where you perform the same experiment with
10 microM FCCP (Uncoupling agent, incubation time 30 min before you added
the dye).
You will be able to measure the difference in membrane potential on
approximately a log . Then repeat the same experiment and fix you cells to
see if the same differences has been maintained, normally CMXRos reacts
with the S-H group of the proteins within the mitochondria and by the way
may give you a picture of the mitochondrial membrane potential. Some groups
say that this work some other not.

Have fun with that !

Patrice

Dr. Petit Patrice X.
INSTITUT COCHIN
INSERM U.567, CNRS UMR 8104, IFR 116
Department of Genetic, Development and Molecular Pathology
Team 5 "Cancer, Apoptosis and Mitochondria"
CHU Cochin Port-Royal
24, rue du Faubourg Saint-Jacques
F-75014 Paris, France.

Tel: 33 01 44 41 24 11
Fax: 33 01 44 41 24 21
E-mail: pxpetit at zeus.cochin.inserm.fr
http://www.cochin.inserm.fr

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