Craig.Turner at nbs.nhs.uk
Mon Sep 4 03:57:34 EDT 2006
Howard your comments on JC-1 are interesting. There are groups in the
platelet field reporting JC-1 ratio for mitochondrial potential and
currently in our lab a colleague is working with a method based on the
published work. His major problem seems to be getting consistent
fluorescence between batches of dye and I don't think he has nailed down a
storage condition he is entirely happy with. All of which means the ratio
becomes unreliable as a comparison of mitochondrial potential over time.
Does anyone else have this sort of problem or any hints I can pass on to
improve the process?
Craig Turner PhD
Scientific development supervisor
Components Development Laboratory
From: Howard Shapiro [mailto:hms at shapirolab.com]
Sent: 31 August 2006 01:38
Subject: Re: JC-1 Dioc6
Paul Waring wrote:
I have been out of touch with the mitochondria field for a few years. Would
anyone care to give me an update on the relative advantages of JC-1 versus
Dioc6 (or other dyes) in measuring mitochondrial membrane potential ?
JC-1 has been found useful for discriminating energized from deenergized
mitochondria, but, as far as I know, has never been demonstrated to
*measure* mitochondrial potential. The distribution of green vs. red
fluorescence is too broad in all directions to make a ratiometric procedure
It is claimed that DiOC6(3) at very low concentrations (around 1 nM) *can*
be used for quantitative measurements (Rottenberg H, Wu S: Quantitative
assay by flow cytometry of the mitochondrial membrane potential in intact
cells. Biochim Biophys Acta 1404:393-404, 1998). I suspect that other
cyanines would also work at this low concentration. If the mitochondria are
energized, the dye concentration inside will be at least 100 times the
external concentration; if the external concentration is much higher than 1
nM, there will be substantial quenching of intramitochondrial dye
fluorescence, and also more interference from dye in the cytosol. At the
lower concentrations, mitochondrial fluorescence would be expected to track
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