The literature I've read seems to indicate mitoSOX is only used with living cells. I'm interested in probing for superoxide anions within the mitochondria of several necropsied organ tissue types. The tissues have been retrieved from mice.
If the answer is no, do you have any suggestions for a more appropriate probe?
Thanks for your help!
Hmmm..last time I had basic chemistry the superoxide anion was not stable. And yes, MitoSOX actually 'works' in frozen tissues...as in producing and artifact (don't ask how I know).
Constructively:
You could use an antibody that detects superoxide reaction products in your tissue; such as in form of modified proteins and lipids. Those may be preserved in frozen tissues, but not sure about their presence after cross linking by PFA. In general this approach is not quantitative and not very sensitive and reliable either.
Good luck, and if it works send PM for co-authorship ;-)
Dear Holt, as Adrian said it is impossible to fix reactive oxygen species themselves. So all ROS-sensitive fluorescent probes could be used only in live cells. But in fixed cell or mitochodnria you can evaluate the consequences of oxidative reactions. For exmaple, you can measure levels of MDA-products, carbonilated proteins, 8-oxoguanine e.t.c.
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