Has anybody immunostained live spinal cord without fixation?
A whole cord? A slice? You mean like after a slice recording? I think most people fix. What are you trying to achieve? Why do you want unfixed, live tissue?
Some people use IB4 staining on live cultured DRGs to identify IB4+ neurons @dmolliver
That was a misunderstanding from my side. Actually the cord is isolated from formalin-fixed carcasses. Do you have any comment on that?
so you have a formalin fixed spinal cord segment? You need to section this somehow and then do standard immunohistochemistry.
The standard process for most of us is:
- Fix animal with PFA (not formalin, but they’re roughly equivalent IMO)
- Cryoprotect with sucrose 30%
- Make cryosections on a cryostat, either free floating or slide-mounted
- Standard IHC immunostaining