Cardiac tissue, fixed in 4% PFA and OCT frozen. Cryosectioned into 10 um sections. Just want to see total mitochondria, does not matter if it is functional or not.
Antibodies against VDAC (aka porin) or citrate synthase could be good options for visualizing mitochondria. Good luck!
Best regards, Christian
Dulguun,
How about Mito Tracker?
http://www.lifetechnologies.com/us/en/home/references/molecular-probes-the-handbook/probes-for-organelles/probes-for-mitochondria.html#head2
good luck,
Michael Delannoy
@Michael: Mitotracker dyes are commonly used for staining mitochondria in living cells. Is it really feasible to use them before cryosectioning of tissue? Are they retained in the sectioned sample?
Best regards, Christian
Christian,
You are correct in that MitoTracker dyes are commonly used for active mitochondria, I would suggest using MT on live cells, fixing then going to the cryostat sectioning.
cheers,
Michael Delannoy
I have used Mitotracker on paraffin sections of mouse heart - it looked as if it had stained the mitochondria. I've also used succinate dehydrogenase histochemistry on unfixed frozen sections to demonstrate mitochondria in muscle cells.
COX-IV (complex 4) works well for mitochondria specific staining. I have seen nice labeling with immunofluorescence on PFA-fixed tissue.
Best,
KE
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