UV-induced Apoptosis on isolated mitochondria, can it be done? - (Jun/16/2011 )

Hi,

As my topic title has said, can UV-induced Apoptosis on isolated mitochondria be done? The reason for this question is that I have to induce apoptosis and isolate mitochondria for AFM imaging.
So if my original plan cannot be done, would the reverse works say that I induce apoptosis, lyse the cell, fix (4% paraformadehyde + 0.1% glutaraldehyde) the mitochondria at certain time interval, then isolate the mitochondria?

Thank you for reading and your kind replies.

I would say that you would have a much better chance with the second option. I am not sure that you could induce apoptosis of the mitochondria isolated from the cell; surely you would need other cellular pathways to trigger it.

Thanks for the reply. What do you think if I try to isolate the mitochondria and add in recombinant Bax protein to achieve the mitochondrial outer membrane permeabilization effect, instead of trying to trigger apoptosis?

That sounds possible, but will take quite a bit of optimisation to get working, I would have thought.

Anybody has any idea how to fix mitochondria on mica. Although fixing is not difficult just by depositing mitochondrial solution on to mica but my problem lies somewhere with AFM imaging. Once mitochondria is attached to mica it gives me a height of 300-400 nm. For such a height its very difficult to use fast mode AFM to study dynamics. Is there any other way around to fix the mitochondria sample on glass or mica so that height becomes less than 200nm.
For example, I tried to deposit mitochondria on compact disc having holes of approx. 200nm height but unfortunately mitochondria did not attach to the disc.