Bone is very autofluorescent, so most dyes that excite in the visible range also excite substances present in bone itself, making it hard to find the specific signal generated by the dye-conjugated antibody. However, I noticed when using DAPI, an intercalating DNA stain used to light up the nuclei, that the background is almost totally black, meaning that bone isn’t autofluorescent in the UV range, where DAPI excites. So, I figured that all I would have to do is find a secondary antibody conjugated to a dye that excites in the UV range, and I should be able to do IHC with little background.

Continue reading →