problem with the mititracker staining - (Feb/13/2008 )
hi
i am facing problem with the mitotracker staining of tissues....
i use mitotracker red 580 at a working conc of 100nM-200nM and image it with 543 nm filter at 63X
and i dont see any staining of the tissues 
..... though the protocol seems to work pretty fine for cells....m finding it difficult to troubleshoot for the tissues
i first fix the tissue in PF then wash n then stain it with mitotracker for 20 min at 37C wash it again followed by acetone permeabilization for further usage....
m planning to fix this time with acetone n methanol after staining.....
plzzz advise me.....
thanking u 
I thought mitotracker was for staining live cells. I didnt know they worked on tissues. You will have to use some mitochondrial antibodies for tissue staining like pyruvate dehydrogenase or some other antibody against protiens in the oxidative phosphorylation.
Hi Scolix...
i know that mitotracker stains live cells.....but then it is also used to stain fixed cells.....what i do is as follows: i take out tissues right after dissection and fix them,,....n then further process.....where does the problem lie then???
can i first stain the tissues right after dissction n then fix cum permeabilize wid acetone??
I am sorry, I was not aware of mitotracker used to stain fixed cells.
try to stain with mitotracker before fixing. you could permeabilize it later.