I am trying to do somework about mitochondria injection, facing lots of difficulties about mitochondria preparation. I prepare the mitochondria using the method from the article of Frezza et al. 2007 of Nature Protocols vol 2 n2.After I resuspend the mitochondria pellet, I got the mitochondrial suspension with large parts, which will block the microinjection needle. When I using mitotracker green to stain the isolation mitochondria, I only see the fluorescence from the large parts, which means there is no mitochondria dissolved in the injection buffer. It bothers me for a long time.I don't know why.
I think that this is a common problem.
You can find a possible solution which includes additional steps (injection into larger cells and viral-mediated membrane fusion of mitocytoplasts) in this article:
Article Transferring isolated mitochondria into tissue culture cells
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