[Histonet] Free radical detection and cell death IHC

[Fabrice GANKAM]

 Sorry to post this again but had problems posting messages lately.

 

Here is my problem, We do not have cryostat in our facility and I will like
to test free radical production in situ in rat brain tissue

Some of the methods involves homogenate of whole part of brain and assay
with component reacting with free radical that does not give you the
geographical distribution of free radical.

I'm therefore looking for a methods involving IHC or IF  in paraffin
section.

I checked for hydroethidine but it seems like the fluorescence fades with
paraffin processing.

 

Has any one used hydroethidine with paraffin processing ?

Does anyone know another method of free radical or oxidative stress
detection on paraffin section ? 

 

We also wanted to asses cell death by some marker with the same tissue
(paraffin processed) the caspase stain is faint and we will like to find a
selective marker of cellular death or irreversible damage (apoptotic or non
apoptotic) that could be used in IHC and IF. 

 

Has anyone ever used PI on rat tissue after paraffin processing ?

Any other marker ?

I heard about the hydroxyprobe but does it work on paraffin embedded tissue
?

 

Thanks for your help guys

 

Fabrice