I am looking for a staining method to label dead neurons in the ischemic mouse brain.I use coronar cryosection from mice which were subjected to transient MCAo and reperfusion. I tried NeuN+TUNEL but unfortunately the dead neurons in the ischemic hemisphere lose too much NeuN.
Do you have any suggestions or even a protocol?
Thanks in advance!
Thank you Guillaume! I've also thought of FluroJade B staining, however, I was not quite sure if I can put cell death and degeneration on the same level. But since I am not staining for cell death in a degenerative model, it should be fine.
Hi Anne-Sophie, have you seen the vanadium acid fuchsin staining that exploits the acidophilia of dying neurons? Maybe you can have a look at the protocol below...
But in general TTC and Fluoro-Jade B stainings are the most used, as Guillaume already pointed out. The Tunel has always been a mystery for me, sometimes it works, sometimes it doesn't and I cannot make a rational sense out of it...
Good luck!
http://www.sciencedirect.com/science/article/pii/S1385299X00000040
If you are looking for focal ischemic changes TTC staining gives you clear idea of infarct area but for global ischemic changes TUNEL/FluoroJade/H&E/etc will be helpful.
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