Hello,
I am performing interneuron electrophysiological recordings in 300-um thick mouse hippocampal sections. I am filling them with 0.3% biocytin and want to reconstruct both the dendrites and the axons for subsequent analysis. However, streptavidin Alexa488 does not penetrate well enough into the tissue for axonal recovery. I've tried to re-section them on a cryostat but with no luck.
Any suggestions?
As anyone tried just putting a non-conjugated Alexa antibody into the intracellular solution? Would switching to a DAB-based post-hoc staining method provide better axonal recovery?
Dendrites look great but axons cannot be seen.
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