Re: Mounting tissue section onto the coverslip

Posted by Glen MacDonald on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Mounting-tissue-section-onto-the-coverslip-tp7591021p7591029.html

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Coverlsips can be charged or else subbed, just as are slides. Several methods have been mentioned in this thread,others may be found in the literature or hjstology methods texts.

I’ve always used, and trained others, to mount on slide, if processing through clearing agent,  don’t let the slide dry out.  that encourages bubbles trapped within voids in the section.  Apply mounting medium along one edge of coverslip  (usually the edge furthest from me parallel to the slide ).  hold coverslip at an angle, 30-60 degrees,using forefingers to the rear and thumbs on edge closest to me , bring the back edge with medium down to the slide, then slowly lower the coverlslip down ono the slide, controlling with your thumbs.  Allow the wavefront of mounting medium to displace any air.  Then I flip the slide over onto a kimwipe to blot excess mounting medium for a few minutes.  The weight of the slide presses out excess.  then flip back upright and allow to harden. Often a bubble can be “massaged” out by gently pressing on the coverslip towards the nearest edge.

Howecver, if you are seeing a mass of fine bubble-like thngs appearing a day or so after coverslipping  sections cleared through a solvent like xylene, they may indicate insufficient dehydration and those are residual water droplets.  

Glen

> On Jun 22, 2020, at 1:20 PM, Craig Brideau <[hidden email]> wrote:
>
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>
> Could this be due to the slides being charged, while the coverslips are
> not? Most of the slides I have used are charged to encourage the tissue to
> stick. Otherwise some sort of surface preparation or pre-coating may be
> necessary to promote adhesion.
>
> Craig
>
> On Mon, Jun 22, 2020 at 2:05 PM Erika Wee <[hidden email]> wrote:
>
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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
>>