I would like to mount some tissue sections that contain a lipophilic dye that bleeds with glycerol-based mounting media. Do you have any favorites? Antifade reagents?
Some dyes (for example- Flouro Jade) are incompatible with glycerol based or aqueous mounting media. You can avoid these media. Instead one can use styrene based mounting medium- for example DPX.
To the best of my knowledge Vectashield is glycerol based medium just like Permafluor.
Citifluor provides different mounting media, which are glycerol-free, like e.g. AF3 (PBS based) but they are non-hardening!
DPX or other xylene-based low fluorescent mounting media are wonderful with fluorescence in that the completely clear the tissue and are permanent I have extensively used Krystalon with great success. The beauty of use of xylene based media is that if sections are very dry and (I usually minimize lower alcohol concentrations and lengthen time in 100% and xylenes) then they not only do not fade much (some fluorophores are better than others) but remain fluorescent for years (>10). If TSA amplification is used our sections remain bright forever.
My favorite is ProLong-Gold from Molecular Probes/Life Technologies. It works great with any red and far red flourophores, as well as all nuclear staining I tried so far (DAPI, ToPro-3. It is excellent to avoid photobleaching. Some people had issues with the GFP but, now they developed ProLong-Diamond that works with green. I myself using Alexa 488 didn't have that problem.
However, you need to be aware that ProLong need to be cured prior to imaging to reach a near-perfect optical path. I usually coverslip the slides and keep them overnight in a drawer to set firm (obviously a designated drawer, so, NOBODY will be opening or putting stuff there when I am not watching). Some people find this cumbersome because you can't image ASAP. But, once it sets hard the slides stay still for much extended time that other mounting media.
Also, Mol Probes recently improved the bottle to a dropper because it is very sticky, so, you need to plan to spend some extra time to SLOWLY & CAREFULLY coverslip your slides. But, you don't need to close the coverslip with nail polish plus, you don't need to refrigerate the slides.
Lastly, your images will look awesome!
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