Re: Mounting tissue section onto the coverslip

Posted by Rhonda Powell on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Mounting-tissue-section-onto-the-coverslip-tp7591021p7591026.html

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Erika,

There are definitely times when I wish the tissue could be adhered directly to the coverslip, but as you said, the issue is typically that there is no charge on the coverslip to help the tissue stick. Even if it sticks initially, it’s likely to fall off during processing/staining steps. Plus, the fragility of the coverslip means it might break and then you have other issues.

Plus, if you actually try cryosectioning, you start to realize how much skill it takes to get the tissue on the slide, and you have a much bigger surface area target on a slide than a coverslip- people who do this successfully have completely earned my respect!

If the bubbles are the main issue, I would have them practice their coverslip technique, even with no tissue on the slide. Someone else just gave a good technique and I’ll add that I use the back of a pipet tip to gently and carefully squish out any bubbles before I seal the coverslip or allow the sample to dry in something like ProLong Gold. A little patience goes a long way!

If they have wrinkly tissue sections, there are other remedies for that, but it sounds like that is not what you are describing.

Rhonda

Rhonda Reigers Powell
Research Lab Manager
Clemson Light Imaging Facility
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From: Confocal Microscopy List <[hidden email]> on behalf of Erika Wee <[hidden email]>
Sent: Monday, June 22, 2020 4:04:59 PM
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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