Hello,
For those with experience with fluorescence immunohistochemistry in tissue sections, I am looking for recommendations for a mounting medium that is hardening, so that I can avoid use of plastic sealant around sides of coverslip. Any recommendations for a specific brand that you found works well in terms of hardening and preservation of fluorescence signal (please provide catalogue number if possible)?
Thanks,
For Immunofluorescence of tissue sections our lab has a lot of success using the Vectashield hardset mounting medium (H-1400) which is easy and ready to use out of the box. In terms of preservation, I've had slides stored away for roughly 3 months and the signal is still able to be observed.
I wouldn't recommend Vectashield Hardset as I have found that it outgases, forming bubbles under the coverslip, and it has blue autofluorescence.
Instead, I would recommend Prolong Diamond, which cures and doesn't need secondary sealing and has superior antifade. There is also ProLong Glass, which is the same but has an RI identical to glass, giving best resolution, particularly for thick samples.
Full disclosure: I work for Tech Support for the company that sells those and helped develop our antifade mounting media when I was in R&D and as an adviser on the projects after moving to Tech Support.
We offer the Vectashield Vibrance, a hardening mounting medium which resolves issues with blue autofluorescence and bubble formation found with the Vectashield HardSet. This new formulation has been benchmarked against competitor's brands with equivalent or superior signal retention.
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