Posted by
Sylvie Le Guyader on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Mounting-tissue-section-onto-the-coverslip-tp7591021p7591030.html
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Hi there
* We advise our users to always put their sections on the coverslip, not on the slide. The aberrations start showing only a few um from the coverslips with reasonably high NA objectives.
* The bubbles come from how you apply the mounting medium, not from the thickness of the glass you use. I agree with the advice that others gave in this thread. Not using a squeeze bottle to apply the mounting medium helps. Use a glass rod dipped in the medium to transfer it.
If this is not possible, one needs to carefully flip the squeeze bottle, wipe the first drop of medium out and apply the following drop of medium on the slide/coverslip without allowing air back in the bottle.
* Cleaning the coverslips with 1M HCl greatly helps sections to attach. See this video.<
https://www.youtube.com/watch?v=j5_JsPPQTxo>
* To help collect sections (or to cytospin) on the coverslip, simply tape the coverslip on a slide with masking tape. This has the added advantages that you can store the coverslips in a slide box as you would do with slides and that you can write notes on the slide.
* This video<
https://m.youtube.com/watch?v=cDQFG-Pdm0o&feature=youtu.be> shows how one can collect paraffine sections using a coverslip.
* I have not tried with sections but this paper<
https://pubmed.ncbi.nlm.nih.gov/30923225/> (suppl methods) describes how one can use liquid glue to stick beads to a coverslip. It would be good to try with sections and see if it helps the tissue adhere during rough antigen retrievals. If anyone tries, please let us know! :D
Med vänlig hälsning / Best regards
Sylvie
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Sylvie Le Guyader, PhD
Live Cell Imaging Facility Manager
Karolinska Institutet- Bionut Dpt
Blickagången 16,
Room 7362 (lab)/7840 (office)
14157 Huddinge, Sweden
mobile: +46 (0) 73 733 5008
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Subject: Re: Mounting tissue section onto the coverslip
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Hi Rhonda,
Thanks very much for your quick response.
Our histology core usually mount sessions onto the glass-slides for the labs, which I can understand, and they were sectioned and mounted beautifully, but the students who was working on post staining and mounting, they use big cover-slip to cover many tissue sections and put uneven mounting media in between, also you can see there are bubbles everywhere.
Some labs here are doing their own cryosections in lab, and they told me the cryosectioned tissue won't stay onto the coverslips.
This is fine if they are using 10x or 20x to do whole section imaging, but for RNA-FISH or for higher resolution imaging, mounting the section to the coverslip become more critical.
Best,
Erika
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