Our lab is looking to measure mitochondrial dysfunction in spinal cord tissue. We think that a drug is reducing mitochondrial damage normally caused by experimental autoimmune encephalomyelitis. The spinal cord tissue is ex vivo and is fixed with PFA. What is the best way to go about getting some preliminary data to determine if our drug is actually reducing mitochondrial dysfunction? I would prefer to use immunohistochemistry if possible. But doing a western blot is also an option.Thanks!
Hi,
I think you could measure mitochondrial membrane potencial. The probe that I know that does that is JC-1, but looking at some papers I've seen that there are some others...
Article Mitochondrial membrane potential probes and the proton gradi...
Chapter Mitochondrial Membrane Potential Assay
Article Flow Cytometric Analysis of Mitochondrial Membrane Potential...
I hope I've helped you!
Best regards,
Jordana.
Dear Meyer,
On the basis of staining as well. Mitochondrial when get calcified show the dense deposits they considered dead.
Best,
Mannan
In PFA fixed tissue... sorry.. too late...
Most of markers in the first answer (e.g. JC1, TMRM, ...) does not work in PFA fixed tissue and most also just in cels. Some say are fixable, that means 1) stain 2) then fix, not the reverse.
What I would is to restart again and isolate mitochondria from the spinal cord, then do mitochondrial physiology.
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