DAPI staining

Hi all,

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?



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