DAPI on top of HE - (Oct/17/2007 )
i did the HE staining with all dehydration steps and then rehydrated it back and coverslipped with prolong antifade with DAPI. HE staining is OK, but i don't see any DAPI fluorescense. Is is because dehydration/rehydration steps or what did I do wrong?
What are actially the rules of fluorescence on the regular staining?
Also, I've posted a guestion about the image-pro software in general Lab forum. Could you help me with that too?
Thanks a lot in advance.
The "H" stains the nucleus very dark color. You fluorophore is not longer be able to bind to DNA and generate fluorescence.
Thanks, GeneHunter!!! I kinda guessed that, but thought that they have different binding sites. Thanks a lot!!!!! So if I do only the Eosine staining, then rehydration steps ant then coverslip with prolong DAPI - would I get the pink and blue?
Eosine should be fluorescent as well, although I am not 100% sure.
yeah, it gives a little bit of non specific fluorescence.