Home-made Anti-bleach (?), vectashield and Immunocytochemistry - (Nov/08/2012 )
I have some ideas regarding immunocytochemistry.
They are more like questions actually.
Is there a good way to protect samples from bleaching? Samples with a bound flourescent secondary antibody?
I used to use vectashield in another lab but here everyone is using glycerol/PBS. Will there be a diffrence?
Can I add something to make it "vectashield-ish"?
From what I understtod vectashield are cheap chemicals sold expensively...
PS. And do I need to bother at all?
Keep the slides in the dark at 4 deg C or lower (frozen even). Slowfade and slowfade gold work well for me.
Good idea, realised (checking, after hearing you) that 70% glycerol would not freeze at -4*C. Still though Im not sure about the integrity of my small worms :/
At any rate I´ll start looking at the company products you mentioned.
Sigma also have its own product: Fluoroshield
But as all of them are quite expensive. At least this one (I use it) is a mounting medium rather than just a antifading reagent. It may contain some polymer based on the plastic/silicone-like residue it left on the rim of the bottle.
I don't know about your worms but most fluorescent-stained stuff is recommended to be stored at -20° C rather than 4° C, though the later is common for short term storage: examination the next day or so.