Spinal cord staining without fixation

Experimental Techniques Protocols
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1

Hello everyone,
Has anybody immunostained live spinal cord without fixation?

2

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

3

That was a misunderstanding from my side. Actually the cord is isolated from formalin-fixed carcasses. Do you have any comment on that?

4

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:

  1. Fix animal with PFA (not formalin, but they’re roughly equivalent IMO)
  2. Cryoprotect with sucrose 30%
  3. Make cryosections on a cryostat, either free floating or slide-mounted
  4. Standard IHC immunostaining
  5. Image