JC-1 observation and question

From: David Gebhard (gebhard@aecom.yu.edu)
Date: Thu Feb 10 2000 - 18:30:51 EST


Greetings,  

	I have an investigator who sent the note below to me after we together did
the FACS and fl. microscope observation......
	The cells treated with valinomycin were nicely depolarized and showed a
stong green (little orange) signal.  The cells in question were bright
orange and indeed full of bright orange mitochondrion.

	Together we are stumped...hence the note to , as we say here in Da
Bronx.....Da List!

>
>I'm using JC-1 to look at the time couse of mitochondrial specific events
>in drug-induced apoptosis. Basically, I treat my cells with drug for
>various time points (Up to 24hrs maximum), after which I harvest adherent
>and non-adherent cells and stain for 15 min at 37 degrees in 1uM of JC-1.
>Instead of depolarisation (a change from orange aggregate formation to
>green fluorescence), I see a hyperpolarization ....my drug treated cells
>are bright orange (even more than controls), reaching a maximum intensity
>at the 24 hr time point. These cells I know to be 80 - 90 % apoptotic, and
>I can even see a classic apoptotic morphology (blebbed membranes) with the
>bright orange mitochondrial staining.
>
>Since the drug I'm using is very hydrophobic could this be having an effect
>on the mitochondrial membrane potential alone...or could it somehow be
>accumulating in mitochondria?
>
>Any suggestions would be gratefully appreciated.
>
>
>Hayley McDaid, Ph.D.
>Molecular Pharmacology
>Golding Room 201
>Albert Einstein College of Medicine
>1300 Morris Park Avenue
>Bronx, New York 10461
>
>Telephone: 718-430-2192
>Fax: 718-430-8922
>E-mail: mcdaid@aecom.yu.edu
>
>
>
>
Dave Gebhard--Director FACS Facility
Albert Einstein College of Medicine
Chanin Building 309
1300 Morris Park Avenue
Bronx, New York 10461
718-430-2724/ 3573  FAX 718-430-8550



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