Hi IHC and IF nerds,
I am having some trouble with my IF staining. I have been staining cells with FITC, AF555, DAPI and AF647.
I have what looks like a green haze covering the whole of my sample in the green channel, however i can see my specific staining underneath this. Does anyone have any ideas what could be causing this? I have mounted with prolong gold and the problem doesn't occur straightaway, usually after a couple of days post staining. Has the FITC dye possibility bled into the mountant? has it gone off due to slides being left out at RT?
any advice would be great.