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DAB & DAPI together for ihc? - (Jan/21/2010 )

Does anyone know if you can double stain tissue (brain) using DAB to visualize the antibody stain and DAPI to visualize the nuclei? Only because we already have DAPI available in house...


You realise the DAB is for standard transmitted light and DAPI is fluorescent?


Yes, but i figured it would be easy enough to turn the fluorescent light on and off and overlay on another picture...


So long as the DAB won't autofluoresce or interfere with the DAPI fluorescence, then it should be OK. Be aware that heavily fixed tissue (e.g. formaldehyde fixed) has a lot of background fluorescence due to the aldehydes left behind.


Have you tried and how was that?


for me, dapi(vectashield) and cfos-DAB didn't go well together... where dab stained seen dark in dapi.


maybe dab pigment opaqued dapi emission light for they stain cellular nucleus(because rather for it was cfos than dab itself) together


Insofar as the DAB won't autofluoresce or meddle with the DAPI fluorescence, at that point it ought to be OK. Know that intensely settled tissue (e.g. formaldehyde settled) has a great deal of foundation fluorescence because of the aldehydes abandoned.