I have processed sciatic nerve as usual (4% PFA followed by increased gradients of sucrose and sectioned in cryostat) with very good results, however I started with vagus nerve from rat and the same procedure doesn't work, It seems that the myelin is destroyed and very poor hystological quality in general. I hope somebody has suggestions or could share your protocol.
Thanks!!!
Hi-- I suggest you minimize fixation time (approximately 10 mins) following a perfusion with PBS (approximately 50mL). Neve fibers can be difficult to image. Did you switch species? (mouse to rat?) This could be an antibody issue if so.
I have attached a useful paper that describes staining of NMJs and nerve fibers. Their procedure is very effective and could possibly be applied to your needs.
Let me know if you need antibody suggestions!
All the best,
Elliot
Hello Eliot, thank you so much for your answer, I didn't change the specie, just different nerve, I will try with your suggestions. All the best
Alejandra
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