Hi there,

Why don't you want to use a fluorescent staining like mito tracker? If you are equipped for fluorescence miroscopy there shouldn't be any problem is the fluorophores have significantly different excitation and emission wavelengths.

You can use Mitotracker-Red from molecular probes, which will not interfere with your GFP tagged probe. But make sure the tissue is fixed well for the stain to work. 

Probes like mitotracker will work for live mitochondria. As far as I know, in fixed cells, the mitochondria loose their membrane potential. Hence, they will not be able to retain the probe. 

Hi again,

So the easiest is to use a suitable mitochondria specific antibody as a primary antibody (a quick search gave me this : http://www.novusbio.com/ACADVL-Antibody_NBP1-54378.html)  and the convinient (non FITC) fluorescent secondary antibody (you should find something OK from there for instance: http://www.pierce-antibodies.com/targets/t/DyLightFluorSecondaryAntibodies.cfm).

No, if you will look at the Mitotracker dye from Molecular probes. The probes have thiol-reactive chloromethyl moiety that keeps them adhered to live mitochondria post fixation. I have used them myself. Only fixation step needs to be taken care of. 

All the best!

I agree with Mamta. Several types of MitoTracker (including Deep Red, which should work fine with your GFP signal) are suitable for both live-cell and fixation fluorescence. You should check out the information and protocols on Life Technology! https://tools.lifetechnologies.com/content/sfs/manuals/mp07510.pdf 

Why don't you want to use a Mn-SOD or COX IV staining?

Try a-Tom20, should work

You can use JC-1 staining which would detect the mitochondrial membrane potential.

Is it possible to use the mito tracker for tissues ?

I need to compare mitochondria density between 2 different tissues... Anyone can suggest me a method?

thanks!

@ Olivia harding, the information provided at the link suggested by you endorses the fact that Mito tracker cmx ros is capable of keeping the cells stained even after fixation. It no where says of its ability of staining pre fixed cells/ tissues. In fact the whole idea of staining depends upon the potential on mitochondrial membrane. Please give reliable information, with pin pointed approach and name of the reagent if you intend to help a researcher. Thank you.

For clarification, MitoTracker works great in live cells, and in live cells that are treated with MitoTracker and subsequently fixed. If you have cells/tissues that are already in fixative, MitoTracker will *not* work. Your best bet is to use antibodies, as mentioned above.

Vikram Narayan

My question is significantly off topic, but it bothers me a lot..

Given that mtDNA can be stained in living cells with EtBr, Pico, etc., why can't nucleoids in fixed cells (PA and EtOH) be visualized with the same dyes? I know that mitochondrial potential is lost, but the mtDNA is still there, and logically it should be possible to visualize it with a dye such as Pico green, for example, which is highly specific for dsDNA and intercalates.