Hi everyone,

            I am trying to set up a protocol for counterstaining emulsion dipped slides (for in situ hybridization using 35S labelled radioprobe) with cresyl violet stain. I tried to do the same using 1% cresyl violet acetate for 15 min followed by dehydration in serial 50, 70, 80, 95 and 100% Ethanol and dipping in Xylene followed by Permamount. The brain sections on my slide have the in situ grains intact, however the cresyl violet stains the cells, especially the neurons on the sections very lightly. I am not sure if this is due to the emulsion , dehydration or some problem with my staining protocol. I would appreciate it , if someone on this list has tried this experiment before and can help me troubleshoot ( or perhaps post a protocol for the same).

Thanks in advance,

Sanjay