JC-1 dye is a sensitive marker for mitochondrial membrane potential change. Generally, it gives Red fluorescence, but in case of mitochondrial membrane potential change, the dye gives green fluorescence. As far as I know, Mitotracker Red and JC-1 are the same class of dyes and the fluorescence of MitoTracker Red is mitochondrial membrane potential-dependent. Have anybody used MitoTracker Red to detect membrane potential change through microscopy or flow-cytometry?
Thanks is advance!
I did not use Mitotracker Red dyes for measuring membrane potential (instead I used JC-1). Therefore I can't share my experiences. Instead I attach a protocol that utilizes Mitotracker Red CMXRos for assaying the mitochondrial membrane potential. Hopefully the information is useful to you.
Regards, Christian
Technical Report Detecting changes in the mitochondrial membrane potential by...
Dear Soumajit,
we use in our group different mitochondrial dyes for different readouts. We don't have the Mitotracker Red but we used Mitotracker green and deep red for staining of mitochondria in live cells. Some of them load along the potential but do not diffuse out freely when the potential drops.
As a readout for Mitochondrial membrane potential we use Tetramethylrhodamin-methylester (TMRM) in red and Rhodamine123 in green. Both dyes can freely diffuse in the cell and accumulate in the mitochondria due to their positive charge. When the potential drops the Dye diffuses out of the mitochondria again.
While in my hands Rhod123 loaded also in other organelles especially in the kidney in vivo, TMRM is highly specific for Mitochondria. So far I used TMRM in 2 different kidney cell lines, acute kidney slices and in vivo. Another plus for it is its cheap price compared to the Mitotracker dyes. I never quantified the changes but used it as general measure for mitochondrial health and it reacts to toxins like FCCP.
Regards,
Claus
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