I have been trying to stain with FJC and every time I put my tissue in the Potassium Permanganate, it starts to fold on the slide. Sometimes, it will flatten out once it is incubated in the FJC/acetic acid solution while other times, the tissue comes off the slide. The tissue is perfusion fixed brain in 4% PFA cut at 40um - then mounted onto gelatin coated slides and dried overnight. We follow the protocol provided by millipore and have had limited success - the tissue integrity is so compromised that it is difficult to obtain representative pictures. We also have difficulty counterstaining the tissue with Hoechst at 1:10,000 - added in the FJC solution. Any suggestions would help.
Fluoro Jade C... I think I figured it out - pH of water really matters - and Fluoro Jade B works better in rat tissue than C...
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