My lab wish to establish a protocol for immuno EM of mouse sciatic nerve. Does anyone a an established protocol, or can refer to labs/papers doing immuno EM on sciatic?
So far, we encounter technical difficulties when the fixed tissue is embedded in agarose for cutting (prior to incubation with primary antibody); it wiggles and cannot be held firmly in the matrix. We speculate it is the rigid epineurium that makes cutting difficult. Experimenting with agarose % has not solved the problem.
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