I am trying to look into some GABAergic interneurons in the thalamus. I fix the animal with 4% PFA overnight and then block the tissue in OCT medium and section on a freezin microtome at 40 microns.
While my staining for NeuN has been giving me beautiful results, I cannot say the same about GAB65/67 and Parvalbumin staining. GAD staining is very very weak while parvalbumin does not seem to work. I was discussing this with a coulleague and he suggested me to use glutaraldehyde instead of paraformaldehyde but could not give any reason for it.
I did my research on the internet and since both GAD and Parvalbumin are not membrane associated, I do not understand what can change? Any idea guys on what I can do to improve the results of my stainings.
Poor IHC results...need help