Mounting tissue section onto the coverslip

[Erika Wee-2]

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Hello,

When I train people here to use the microscopes in the facility, 90% of the samples the tissue sections were mounted on the glass slide with lots of bubbles, so I am trying to convince people to mount the tissue section directly onto the cover-slip side.

But many people told me the tissue won't stay stick on the cover-slips. Would you mind to share how you or your lab mount the tissues sections? Do you or your histology core mount tissues directly onto the coverslips? Do you charge or coat the coverslips so the tissue section can stick better for staining?

Any information would be greatly appreciated.

Many thanks.
Best,
Erika

[Rhonda Powell]

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Hi Erika,

Having done a little bit of histology myself, I can say that it’s a lot easier said than done to get tissue sections cut and placed on a cover slip!

If they have issues with bubbles, I feel like that would be true whether they place the tissue on the slide or the coverslip though.

Is your bubble issue with cryosections or paraffin sections? That might help some of us with limited histology experience give some other troubleshooting advice.

I also have some histo friends I could refer your question to if we can’t help!

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 2:45 PM
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Subject: Mounting tissue section onto the coverslip

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Hello,

When I train people here to use the microscopes in the facility, 90% of the samples the tissue sections were mounted on the glass slide with lots of bubbles, so I am trying to convince people to mount the tissue section directly onto the cover-slip side.

But many people told me the tissue won't stay stick on the cover-slips. Would you mind to share how you or your lab mount the tissues sections? Do you or your histology core mount tissues directly onto the coverslips? Do you charge or coat the coverslips so the tissue section can stick better for staining?

Any information would be greatly appreciated.

Many thanks.
Best,
Erika

[Benjamin Smith]

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I can say the trick for me has always been to put a small drop of
mounting media on top of the sample on the slide, and then using a pair of
#5 forceps (or any ultrafine forcep) holding onto one side of the
coverslip.  Holding the coverslip vertically relative to the slide, lower
the coverslip until the side opposite the forceps is resting on the slide,
and then gently lower the side with the forceps until the media first wets
onto the coverslip.  Continue to gently lower the coverslip, allowing the
mounting media to wick along the coverslip.  This way, no air will ever get
trapped under the coverslip.  In short, gradually lower just one side of
the coverslip so that the mounting media can wet along the length of the
coverslip, and you will almost never get air bubbles.

Cheers,
   Ben Smith

On Mon, Jun 22, 2020 at 11:45 AM Erika Wee <[hidden email]> wrote:

> *****
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> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
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> *****
>
> Hello,
>
> When I train people here to use the microscopes in the facility, 90% of
> the samples the tissue sections were mounted on the glass slide with lots
> of bubbles, so I am trying to convince people to mount the tissue section
> directly onto the cover-slip side.
>
> But many people told me the tissue won't stay stick on the cover-slips.
> Would you mind to share how you or your lab mount the tissues sections? Do
> you or your histology core mount tissues directly onto the coverslips? Do
> you charge or coat the coverslips so the tissue section can stick better
> for staining?
>
> Any information would be greatly appreciated.
>
> Many thanks.
> Best,
> Erika
>

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University of California, Berkeley
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Fax [phone]
e-mail: [hidden email]
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[Erika Wee-2]

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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

[Rhonda Powell]

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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
To: [hidden email] <[hidden email]>
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

[Craig Brideau]

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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:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> 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
>

[Tobias Baskin]

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Hi Erika,
             If you have access to a plasma cleaner putting your slides
with sections in a plasma for 30 sec or so just before mounting makes
the slides and sections super hydrophyllic and the mounting medium wets
a treat. I found this trick quite helpful for minimizing bubbles, esp
those that love to form right over the section. Hope this helps. Tobias

On 6/22/20 2:42 PM, Erika Wee wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hello,
>
> When I train people here to use the microscopes in the facility, 90% of the samples the tissue sections were mounted on the glass slide with lots of bubbles, so I am trying to convince people to mount the tissue section directly onto the cover-slip side.
>
> But many people told me the tissue won't stay stick on the cover-slips. Would you mind to share how you or your lab mount the tissues sections? Do you or your histology core mount tissues directly onto the coverslips? Do you charge or coat the coverslips so the tissue section can stick better for staining?
>
> Any information would be greatly appreciated.
>
> Many thanks.
> Best,
> Erika

--
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      /  \  /     / \  /      \      Professor   (he)
     /   / /     /   \ \       \      Biology Department
    / __/ /__   /___  \ \       \__    University of Mass.
   /     /     /       \ \       \      611 N. Pleasant St.
  /     /     /         \ \       \      Amherst, Massachusetts
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[Julian Smith III]

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Thanks, Ben
Exactly the method I've been using for over four decades now.
One addition: if you have giant sections (think whole rat heart), or a
half-dozen rows of serial sections, put a drop of clearing agent on the
section, then add the drop of mounting medium. Depending on how hot your
lab is (or if you're working in a hood), the clearing agent (I use
xylene) can partially evaporate before you get the coverslip down.
Julian

On 6/22/20 3:23 PM, Benjamin Smith wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> I can say the trick for me has always been to put a small drop of
> mounting media on top of the sample on the slide, and then using a pair of
> #5 forceps (or any ultrafine forcep) holding onto one side of the
> coverslip.  Holding the coverslip vertically relative to the slide, lower
> the coverslip until the side opposite the forceps is resting on the slide,
> and then gently lower the side with the forceps until the media first wets
> onto the coverslip.  Continue to gently lower the coverslip, allowing the
> mounting media to wick along the coverslip.  This way, no air will ever get
> trapped under the coverslip.  In short, gradually lower just one side of
> the coverslip so that the mounting media can wet along the length of the
> coverslip, and you will almost never get air bubbles.
>
> Cheers,
>     Ben Smith
>
> On Mon, Jun 22, 2020 at 11:45 AM Erika Wee <[hidden email]> wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> Post images on http://www.imgur.com and include the link in your posting.
>> *****
>>
>> Hello,
>>
>> When I train people here to use the microscopes in the facility, 90% of
>> the samples the tissue sections were mounted on the glass slide with lots
>> of bubbles, so I am trying to convince people to mount the tissue section
>> directly onto the cover-slip side.
>>
>> But many people told me the tissue won't stay stick on the cover-slips.
>> Would you mind to share how you or your lab mount the tissues sections? Do
>> you or your histology core mount tissues directly onto the coverslips? Do
>> you charge or coat the coverslips so the tissue section can stick better
>> for staining?
>>
>> Any information would be greatly appreciated.
>>
>> Many thanks.
>> Best,
>> Erika
>>
>
> --
> Benjamin E. Smith, Ph. D.
> Imaging Specialist, Vision Science
> University of California, Berkeley
> 195 Life Sciences Addition
> Berkeley, CA  94720-3200
> Tel  [phone]
> Fax [phone]
> e-mail: [hidden email]
> https://vision.berkeley.edu/faculty/core-grants-nei/core-grant-microscopic-imaging/
>
> CAUTION: This message originated from an external source

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[Caroline Miller-2]

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You will still get bubbles, just on the underneath if you put the sections on the coverslip.

I put the coverslip flat on kimwipe, then a dot of moutant on the coverslip (you really need to play with the right amount for your application), then wet the moutant with a drop of liquid (either xylene based or PBS, again depending on application), then flip the slide and catch the drop on the slide and let capillary action do the rest.

You can push out small bubbles, but use a tool, not your greasy fingers or moutant smeared gloves, I use a metal probe.

I then wick up the excess with a kimwipe on the edge of the slide

Important! Don't keep smearing the moutant, just start again if it looks like a mess.

Remember smears are the enemy of good imaging!

Caroline



Caroline Miller
Associate Director of Immunohistochemistry and Analytical Histology
Unity Biotechnology

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From: Confocal Microscopy List <[hidden email]> on behalf of Tobias Baskin <[hidden email]>
Sent: Monday, June 22, 2020 1:54:56 PM
To: [hidden email] <[hidden email]>
Subject: Re: Mounting tissue section onto the coverslip

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Hi Erika,
             If you have access to a plasma cleaner putting your slides
with sections in a plasma for 30 sec or so just before mounting makes
the slides and sections super hydrophyllic and the mounting medium wets
a treat. I found this trick quite helpful for minimizing bubbles, esp
those that love to form right over the section. Hope this helps. Tobias

On 6/22/20 2:42 PM, Erika Wee wrote:

> *****
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> *****
>
> Hello,
>
> When I train people here to use the microscopes in the facility, 90% of the samples the tissue sections were mounted on the glass slide with lots of bubbles, so I am trying to convince people to mount the tissue section directly onto the cover-slip side.
>
> But many people told me the tissue won't stay stick on the cover-slips. Would you mind to share how you or your lab mount the tissues sections? Do you or your histology core mount tissues directly onto the coverslips? Do you charge or coat the coverslips so the tissue section can stick better for staining?
>
> Any information would be greatly appreciated.
>
> Many thanks.
> Best,
> Erika

--
       __    ___   ^    ___   ___   Tobias I. Baskin
      /  \  /     / \  /      \      Professor   (he)
     /   / /     /   \ \       \      Biology Department
    / __/ /__   /___  \ \       \__    University of Mass.
   /     /     /       \ \       \      611 N. Pleasant St.
  /     /     /         \ \       \      Amherst, Massachusetts
/     /___  /           \ \___/   \_____   USA  01003    413-545-1533
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[Glen MacDonald]

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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:
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> 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:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> Post images on http://www.imgur.com and include the link in your posting.
>> *****
>>
>> 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
>>

[Sylvie Le Guyader]

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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
@@@@@@@@@@@@@@@@@@@@@@@@
Sylvie Le Guyader, PhD
Live Cell Imaging Facility Manager
Karolinska Institutet- Bionut Dpt
Blickagången 16,
Room 7362 (lab)/7840 (office)
14157 Huddinge, Sweden
mobile: [phone]
LCI website
Follow our microscopy blog!

________________________________
From: Confocal Microscopy List <[hidden email]<mailto:[hidden email]>> on behalf of Erika Wee <[hidden email]<mailto:[hidden email]>>
Sent: Monday, June 22, 2020 10:04:59 PM
To: [hidden email]<mailto:[hidden email]> <[hidden email]<mailto:[hidden email]>>
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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[Erika Wee-2]

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Dear Sylvie,

Thank you so much for the detailed into, and also the YouTube videos. I agree with you 100% that samples should be always mounted onto the coverslip side. I just did a microscopy workshop last week, I find it's really difficult to ask people to modify their existing sample prep protocol to mount the sample to the coverslips.

The YouTube videos are really well made and super helpful, especially I really can't go to each lab to show them how to work on microscopy samples. I just forward the info you provided to my lab and facility users to watch it.

Thank you so much!

Best,
Erika

[Erika Wee-2]

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Dear Rhonda,

Thank you so much for the detailed info, that's super helpful. Sorry if my previous message wasn't clear, there were so many different kind of badly prepared sample I have seen people bringing in to try to image on the facility microscopes.

I realize it's too much to ask and too technical demanding microscopy beginners to mount tissue onto the coverslip, right now I advised them to practice to avoid bubbles as much as they can, but eventually one day if they would like to do higher resolution imaging, it's still critical to mount tissues onto the coverslips.

And yes, the tissues are a bit wrinkled as well, any suggestion would be greatly appreciated.

Thanks again.

Best,
Erika

[Erika Wee-2]

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Hi Glen,

Thank you so much for the detailed info, that really helps me to identify and to troubleshoot the bubbly samples brining in to the microscopy facility.
Especially I understand it's a quite technical demanding and also hard to convince people to modify there existing protocol to mount the tissue to the coverslips.

Many thanks.
Best,
Erika

[Sylvie Le Guyader]

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Hi again

You are very welcome to use our videos.
My experience is that it is easy to convince our users to put their sample on the coverslip. When we train them, they must prepare samples for the training. We simply tell them to make 2 samples in parallel, one the way they normally do it and one identical but on the coverslip. It takes no extra effort so they do it.
Then we show them the difference with a high NA objective and let them decide what they want.
One only sees the effect of not preparing the sample in an optimal way when one looks at the images side by side. 😊

Med vänlig hälsning / Best regards
Sylvie
@@@@@@@@@@@@@@@@@@@@@@@@
Sylvie Le Guyader, PhD
Live Cell Imaging Facility Manager
Karolinska Institutet- Bionut Dpt
Blickagången 16,
Room 7362 (lab)/7840 (office)
14157 Huddinge, Sweden
mobile: [phone]
LCI website
LCI microscopy blog

-----Original Message-----
From: Erika Wee <[hidden email]>
Sent: Tuesday, 23 June, 2020 19:03
To: [hidden email]; Sylvie Le Guyader <[hidden email]>
Cc: Erika Wee <[hidden email]>
Subject: Re: Mounting tissue section onto the coverslip

Dear Sylvie,

Thank you so much for the detailed into, and also the YouTube videos. I agree with you 100% that samples should be always mounted onto the coverslip side. I just did a microscopy workshop last week, I find it's really difficult to ask people to modify their existing sample prep protocol to mount the sample to the coverslips.

The YouTube videos are really well made and super helpful, especially I really can't go to each lab to show them how to work on microscopy samples. I just forward the info you provided to my lab and facility users to watch it.

Thank you so much!

Best,
Erika



När du skickar e-post till Karolinska Institutet (KI) innebär detta att KI kommer att behandla dina personuppgifter. Här finns information om hur KI behandlar personuppgifter<https://ki.se/medarbetare/integritetsskyddspolicy>.


Sending email to Karolinska Institutet (KI) will result in KI processing your personal data. You can read more about KI’s processing of personal data here<https://ki.se/en/staff/data-protection-policy>.

[Sylvie Le Guyader]

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About wrinkly sections:
This takes practice at the microtome.
But in general I talk people out of making super thin sections. I think that making sections as thin as possible is a remnant of hospital practice where one uses bright field images and colour dyes.
I advise people to make sections of over 1 cell diameter so that they do not cut every single cell. It is rare that the scientific question requires cutting every single cell.
Having slightly thicker sections (20um) makes it easier to handle the sample. Sections over 100um can be stained free floating then gently transferred to the coverslip with a kiddie paint brush.

Med vänlig hälsning / Best regards
Sylvie
@@@@@@@@@@@@@@@@@@@@@@@@
Sylvie Le Guyader, PhD
Live Cell Imaging Facility Manager
Karolinska Institutet- Bionut Dpt
Blickagången 16,
Room 7362 (lab)/7840 (office)
14157 Huddinge, Sweden
mobile: [phone]
LCI website
LCI microscopy blog

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Erika Wee
Sent: Tuesday, 23 June, 2020 20:04
To: [hidden email]
Subject: Re: Mounting tissue section onto the coverslip

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Dear Rhonda,

Thank you so much for the detailed info, that's super helpful. Sorry if my previous message wasn't clear, there were so many different kind of badly prepared sample I have seen people bringing in to try to image on the facility microscopes.

I realize it's too much to ask and too technical demanding microscopy beginners to mount tissue onto the coverslip, right now I advised them to practice to avoid bubbles as much as they can, but eventually one day if they would like to do higher resolution imaging, it's still critical to mount tissues onto the coverslips.

And yes, the tissues are a bit wrinkled as well, any suggestion would be greatly appreciated.

Thanks again.

Best,
Erika


När du skickar e-post till Karolinska Institutet (KI) innebär detta att KI kommer att behandla dina personuppgifter. Här finns information om hur KI behandlar personuppgifter<https://ki.se/medarbetare/integritetsskyddspolicy>.


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[Martin Wessendorf-5]

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Dear Erika--

It looks as if all of your responses have been emailed directly to you,
rather than to the list.  (Or to be more clear: I haven't seen any of the
responses!).  Would you mind sharing any protocols for making sections to
stick to coverslips?  I expect there are others out there who would
appreciate that--especially if you're making it work for RNA localization.

Thanks--

Martin Wessendorf
List Manager

On Tue, Jun 23, 2020 at 1:06 PM Erika Wee <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear Rhonda,
>
> Thank you so much for the detailed info, that's super helpful. Sorry if my
> previous message wasn't clear, there were so many different kind of badly
> prepared sample I have seen people bringing in to try to image on the
> facility microscopes.
>
> I realize it's too much to ask and too technical demanding microscopy
> beginners to mount tissue onto the coverslip, right now I advised them to
> practice to avoid bubbles as much as they can, but eventually one day if
> they would like to do higher resolution imaging, it's still critical to
> mount tissues onto the coverslips.
>
> And yes, the tissues are a bit wrinkled as well, any suggestion would be
> greatly appreciated.
>
> Thanks again.
>
> Best,
> Erika
>

[Martin Wessendorf-5]

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Dear All--

It looks as if at least some people received all of the responses to
Erika's post--also, the responses can be found on the web version of the
Confocal List.

Did anyone else NOT receive those responses?  You all should have received
several posts with the subject line: "Re: Mounting tissue section onto the
coverslip" with the sender NOT being Erika Wee.

Feel free to respond to me directly rather than cluttering the List with
this!

Thanks for your help trouble-shooting--

Martin Wessendorf

On Wed, Jun 24, 2020 at 9:15 AM Martin Wessendorf <[hidden email]> wrote:

> Dear Erika--
>
> It looks as if all of your responses have been emailed directly to you,
> rather than to the list.  (Or to be more clear: I haven't seen any of the
> responses!).  Would you mind sharing any protocols for making sections to
> stick to coverslips?  I expect there are others out there who would
> appreciate that--especially if you're making it work for RNA localization.
>
> Thanks--
>
> Martin Wessendorf
> List Manager
>
> On Tue, Jun 23, 2020 at 1:06 PM Erika Wee <[hidden email]> wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> Post images on http://www.imgur.com and include the link in your posting.
>> *****
>>
>> Dear Rhonda,
>>
>> Thank you so much for the detailed info, that's super helpful. Sorry if
>> my previous message wasn't clear, there were so many different kind of
>> badly prepared sample I have seen people bringing in to try to image on the
>> facility microscopes.
>>
>> I realize it's too much to ask and too technical demanding microscopy
>> beginners to mount tissue onto the coverslip, right now I advised them to
>> practice to avoid bubbles as much as they can, but eventually one day if
>> they would like to do higher resolution imaging, it's still critical to
>> mount tissues onto the coverslips.
>>
>> And yes, the tissues are a bit wrinkled as well, any suggestion would be
>> greatly appreciated.
>>
>> Thanks again.
>>
>> Best,
>> Erika
>>
>