Suggestion for histologically identifying liver damage. - TUNEL? Apoptosis? (Aug/01/2007 )

Some mutant mice I have seem to be exhibiting different levels of liver damage in response to a toxin I'm giving them, compared to wildtype controls.

However, this is based on the ApopTag kit from Chemicon. The problem is that
a) apoptosis is a very subtle phenomenon, because it's occurring in only a tiny fraction of cells at any given moment
b) apoptosis is not the only likely response to this toxin, and probably not the most physiologically relevant one

Really, all I want is *some* metric of cell damage, death, or distress. Necrosis would be good. Cell membrane leakage would be good. The main criterion is that it be sensitive enough that there are more than one or two positives per organ cross section. What kinds of stains or kits would folks recommend for this purpose?

Thanks.

-bokov-

you could use antibodies raised in the same species which target caspase3 for apoptosis and whatever is relevant for necrosis then just use a single secondary to lable them both and you will have covered a fair chunk of cell death

dom

-Dominic-

QUOTE (Dominic @ Aug 1 2007, 06:58 AM)

you could use antibodies raised in the same species which target caspase3 for apoptosis and whatever is relevant for necrosis then just use a single secondary to lable them both and you will have covered a fair chunk of cell death

dom

Thank you for your reply. It's precisely the 'whatever is relevant for necrosis' that I am looking for ideas on. What kinds of molecular markers would I be looking for in a cell that's falling apart in a non-programmed manner?

It seems that most people who report necrosis data do morphology of H&E stained specimens... but that's subject to preservation artifacts (e.g. cells shrinking in the formalin solution) that can make even healthy tissue look necrotic. I was hoping there would be something more robust out there.

-bokov-