I am doing antigen retrieval on brain sections. Antigen retrieval buffer is tris-Hcl buffer pH 9.5. Heating the buffer at 80 C for 30mins and cool it for 1hr and then process it for regular immunocytochemistry. I use polyclonal primary antibody, biotinylated secondary antibody and incubate with ABC complex and later on stain with glucose oxidase Ni DAB solution. I use free floating sections for the assay but I have trouble when I mount these sections, there are lots of wrinkles and sometimes some parts are missing. As far as staining goes, I get a clear staining. Do you have any suggestions for this? Have any of you tried antigen retrieval on slide mounted sections?? IF so what would be your coating of the slides. I use gelatin coated slides and didnt work well. All sections got detached from it.