Hi everyone!
I'm searching detailed protocol for teasing-fiber technique for immunohistochemistry. My approach to these technique is self made, with few indication and i need some help, in particular on how to prepare the sciatic nerve before the teasing procedure (best fixation protocol to keep myelin safe, for now i use PFA 4% for 10 minute a RT) and how to settle and paste the collected nerves on the glass slide (i am using nothing to paste the fibers on the glass) and the best methods to tease the fiber.
Thank you for the attention and help!
Thanks for your reply.
I had a look to this protocol and for the first (fixation) and the third part (staining) do not think I have big problems and I made some changes according to my needs, especially for the staining for some antibodies (acetone in place of methanol).
My doubts remain on the teasing part and how to better insulate the individual fibers. Now, once isolated from mouse and fixed the sample, i position the sciatic nerve section on the slide and, using well-sharpened tweezers, i try to divide the fibers. The problem remains the ease with which the sample is moved during this step.
Is there a way to keep it as still as possible?
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