Does anyone know if it works to triple-label mouse brain tissue with DAPI, BrdU, and doublecortin (DCX)? I'm using sections of the hippocampus and the subventricular zone to examine cell proliferation and neurogenesis.
It should work in theory, but in my experience, nuclear stainings are often pretty bad after the HCl for BrdU, which is one of the reasons I switched to EdU. Although I think DAPI is less sensitive to that than the Hoescht I was using at the time. I'm sure someone else will weigh in on that.
It should work, just don't do HCl for the BrdU, just do boiling in citrate ph 6, then do the first IHC for brdu, and then DCX.
I've done it, even with HCl treatment. Should be fine. DAPI is still pretty after 1h 2N HCl at room temp.
if you use hcl treat, you shuold control the time of hcl. of course, you can first strain dapi, after fixed use 4%PFA, then hcl treat.this is useful.
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