I am working with a mouse prostate and I have autofluorescence problems when using the Bouin fixative. In fact, I'm not sure what the origin of autofluorescence is ... it could be the tissue itself and the paraffin. Would anyone know how to solve this problem?
Another problem is that the DAPI dye does not work on the material fixed with Bouin. I have tried to use tissue fixed in 4% paraformaldehyde, and embedded in paraffin, in this case the DAPI works, but the tissue morphology is very bad. That is why I would like to use Bouin on immunofluorescence... because the morphology is much better preserved.
Thank you
State University of Campinas, UNICAMP, Brazil
Hi Raffaella.
It's Julia, from Parada's Lab. To reduce the background and autofluorescence I use 50mM (15 min) of Ammonium choride before permeabilization. Also I use glicine 50mM in the secondary antibody solution. Did you try this protocol?
Espero que ajude. Ate mais.
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