I am trying to do a nerve backfill with neurobiotin (visualized with avidin-fluorescein) and in the same tissue section I would like to do a tryptophan hydroxylase immunocytochemistry (visualized with texas red).
However, when I do the usual tryptophan hydroxylase in a DAB reaction I get amazing results, but no luck with the fluorescent double label, even though I still manage to get great neurobiotin label in the same tissue (using ABC-DAB or avidi-fluorescein).
In short, I get specific neurobiotin-avidin-fluorescein, but no specific tryptophan hydroxylase-texas red staining.
Hope this is clear...

I would try another secondery antibody