Re: JC-1 DYE

Hello,
Personally, I cant stand JC-1.	There are many other dyes that you 
could use.  In general the incorporation of these fluorochomes may be 
influenced by parameters other than mitochondrial membrane potential 
(cell size, PM permeability, efficacy of the MDR pump), results can 
only be interpreted when the staining profiles obtained in different 
experimental conditions are identical to the pattern in the presence of 
CCCP(a depolarizing agent). (castedo et al 2002)
In addition I think that JC-1 is very concentration dependant.  
Titration is crucial.  Profiles and results can vary greatly if the 
wrong concentration is used.
And lastly, confirmation of specific staining by microscopy.

	Smiley, S., et al., Intracellular heterogeneity in mitochondrial 
membrane potentials revealed by a J-aggregate-forming lipophilic cation 
JC-1. Proc Natl. Acad. Sci USA, 1991. 88: p. 3671-3675.
	Chen, L.B., Fluorescent Labeling of Mitochondria. Methods in Cell 
Biology, 1989. 29: p. 103-123.
	Metivier, D., et al., Cytofluorometric detection of mitochondrial 
alterations in early CD95/Fas/APO-1 triggered apoptosisi of Jukat T 
lymphoma cells.  Comparison of seven mitochondrion-specific 
fluorochromes. Immunology Letters, 1998. 61: p. 157-163.	Poot, M. and 
R. Pierce, Detection of Changes in mitochondrial function during 
apoptosis by simltaneous staingin with multiple fluorescent dyes and 
correlated multiparameter flow cytometry. Cytometry, 1999. 35: p. 
311-317.
	Castedo, M., et al., Quantitation of mitochondrial alterations 
associated with apoptosis. Journal of Immunological Methods, 2002. 265: 
p. 39-47.

Hope this helps a little
JM



On Mar 3, 2005, at 7:33 AM, <KurtzJ@usa.redcross.org> wrote:

> Question:  Has anyone out there used the dye JC-1 to look at platelet 
> mitochondrial membranes.  I am interested in knowing what
>  to use as a positive and negative control since JC-1 is both in the 
> FL1 and FL2 channels and also a small amount in FL3.  I have a
>  paper by Perotta that measure the membrane potential using JC-1 but 
> doesn't mention any controls.  I have tried the method described
>  in the paper but see very little fluorescence at room temperature.  
> Any information would be helpful.  Thanks, Jim Kurtz