Hello! I've recently been trying to optimise immunofluorescent staining of mouse meningeal tissue, and I'm having trouble mounting it just because it's so thick.
We use prolong to mount and usually ~10 uL is enough for our 10 uM brain cryosections but even 20 uL seemed to be too little for the meninges since the meninges is much thicker (cover slip wouldn't rest on the tissue properly).
Would anyone have any advice on this? I feel like the process would be similar to mounting thicker cryosections of regular tissue.
I would suggest to use a volume of mounting media appropriate to cover all the area of the coverslip, not only the tissue section, this way your coverslip will rest correctly on your tissue and microscope slide.
Personally I've not had to mount really thick sections, but I've seen other people make a shallow well on the slide by gluing coverslips to the slide. This might help keep the coverslip flat.
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