[Histonet] immunofluorescence mounting medium?

[Collette, Nicole M.]

Hello, Esteemed Histonetters,

I am trying to get nice publication-quality images of my immunofluorescent tissue sections. I am currently using Prolong Gold, and after I let the stuff cure for several days, my 100X oil-immersion images are still smeary, even after taking into account the unevenness of the tissue section. I don't seem to have this problem with colorimetric stains (histological stains, LacZ, etc.), so I am thinking the mounting medium is part of the problem? It seems to me that it never fully cures at the middle of the coverslip… Does anyone have a recommendation for a mounting medium that works better for this purpose? Any advice is appreciated.

Thanks in advance, and Happy Monday!

Sincerely,
Nicole Collette
Lawrence Livermore National Laboratory
collette2 <@t> llnl.gov