Hi,
Please read this article. It may be useful for you
Mitochondrial analysis in living cells: the use of rhodamine 123 and flow cytometry.
http://www.ncbi.nlm.nih.gov/pubmed/2945606
Rhodamine 123 is related to mitochondrial transmembrane potential . One of the earliest changes that occurs in apoptosis is a change in transmembrane potential . Rhodamine 123 can be visualized using flow cytometry & the intensity is linear . Therefore it has wide scope in research on apoptosis .
Thank you Dr. Geethanjali Bunglavil. But when you say that the intensity is linear, does it means that in cancer cells, rhodamine accumulates a lot inside of the mitochondria? Is it because of this phenomena that We observe a decrease in the intensity?
Once, inside the mitochondria, is it true that there will be flouresence quenching?
Hi,
Please read this article. It may be useful for you
Mitochondrial analysis in living cells: the use of rhodamine 123 and flow cytometry.
http://www.ncbi.nlm.nih.gov/pubmed/2945606
Thanks Phannibhushann Meka. That helped me a lot. But, still I have a question.
Inside the mitochondria, is it true that there will be flouresence quenching? I don't find any answer for this question.
Im not sure about fluorescence quenching, but there are some reports that some cancer cells, especially stem-like cells have ABC transporters that can pump rhodamine 123 out. So it appears as if there is less fluorescence in these cells. These cells are thought to be the multi-drug resistant cells.
http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0033253
Rhodamine 123 is a cationic dye and hence localizes in mitochondrial. It can therefore be used to detect mitochondrial membrane potential (MMP). Loss of MMP will result in loss of Rh123 fluorescence. You will rarely see anyone using Rh123 for detecting MMP in recent literature. This is due to toxicity of Rh123. Either TMRE or JC1 are preferred. TMRE can be used in nano molar concentrations (50-150 nM) and is not toxic. At higher concentrations TMRE self quenches.
- Gaurav.
Rhodamine 123 can be used to detect mitochondrial membrane potential dissipation. It is not a specific marker for detecting apoptosis. However, it can determine MMP which is associated with mitochondrial apoptosis or intrinsic apoptosis.
Some "consensual recepice" for apoptosis measurements are here attached.
Best regards
Robert
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