lacZ+ immunostaining

From:Peter Huszthy

Good day to you, all histopathologists!

I have frozen sections of brain tissue, where I would like to show that lacZ gene expression is localized to neurons.
Thus, I would like to perform both lacZ and immunostaining against Neurofilament (DAKO mouse monoclonal) on the same section. Since the lacZ in this case is localized to the nuclei, it would not overshadow the Neurofilament staining, localized to axons.
For lacZ staining I have prevoiusly used 0.2% glutaraldehyde+ 2% paraformaldehyde in PBS as fix.
For the immunostaining with this primary antibody, aceton was good, followed by DAKO`s envision kit with HRP-labelled polymer and detection with DAB+substrate-chromogen. How would I combine the two methods the best way?
Would the lacZ fix work all the way thru or would it probably destroy the antigen? I have very few sections, so I cant do too much trial and error.
Thanks for your help. Peter

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