I have a 2mm thick slice of rat brain, and I'm DAPI-staining it at a 10nM concentration at the moment (because thats the number I kept stumbling across). I've left it for roughly a week now, but I'm unsure how long I should leave it. It appears to vary a lot with staining type.
How long should I leave it, and how long should I wash it? Has anyone tried DAPI?
When I stained for perineuronal nets, I got a lot of unspecific binding on the very surface (not too much background other than that though). Would that be due to insufficient washing perhaps?