Mowiol mounting -> bubbles

[Rusty Nicovich]

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We have a project that involves imaging tissue sections mounted in
home-made Mowiol (PVA) mounting medium.  The coverslip is held away from
the slide with an adhesive spacer, the aperture in the spacer filled with
tissue and medium, and a coverslip applied.

These work great for about a month.  After that, we notice more and more
often bubbles appearing in the mounting medium. These are mostly around the
edge and immediately under the coverslip.

We've tried sealing with nail polish.  This seems to have little effect on
the rate or the extent of bubble formation.

My theory is that the PVA is shrinking and pulling away from the coverslip
and spacer.  If that's the case (or if not) is there a way to prevent
this?  Would another sealing material do the job better? Any tips or tricks
out there?

Thanks,
Rusty Nicovich

[Benjamin Smith]

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One thing I would try would be boiling the aqueous components before adding
them to remove any dissolved gasses.  It is oftentimes surprising how much
gas can be dissolved in water.

-Ben Smith



On Tue, Feb 28, 2017 at 5:20 PM, Rusty Nicovich <[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.
> *****
>
> We have a project that involves imaging tissue sections mounted in
> home-made Mowiol (PVA) mounting medium.  The coverslip is held away from
> the slide with an adhesive spacer, the aperture in the spacer filled with
> tissue and medium, and a coverslip applied.
>
> These work great for about a month.  After that, we notice more and more
> often bubbles appearing in the mounting medium. These are mostly around the
> edge and immediately under the coverslip.
>
> We've tried sealing with nail polish.  This seems to have little effect on
> the rate or the extent of bubble formation.
>
> My theory is that the PVA is shrinking and pulling away from the coverslip
> and spacer.  If that's the case (or if not) is there a way to prevent
> this?  Would another sealing material do the job better? Any tips or tricks
> out there?
>
> Thanks,
> Rusty Nicovich
>

--
Benjamin E. Smith, Ph. D.
Microscopy Specialist, Vision Science
University of California, Berkeley
142 Life Sciences Addition
Berkeley, CA  94720-3200
Tel  [phone]
Fax [phone]
e-mail: [hidden email]
http://vision.berkeley.edu/

[Sylvie Le Guyader]

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Hi Rusty

Putting the section on the coverslip and mounting with the slide goes a very long way. Could that be an option?

Med vänlig hälsning / Best regards

Sylvie

@@@@@@@@@@@@@@@@@@@@@@@@

Sylvie Le Guyader, PhD
Live Cell Imaging Facility Manager
Karolinska Institutet- Bionut Dpt
Hälsovägen 7,
Novum, G lift, floor 6
14157<tel:14157> Huddinge
Sweden
mobile: [phone]<tel:+46%20(0)%2073%20733%205008>
office: [phone]<tel:+46%20(0)%2008-524%20811%2072>
LCI website

---- Rusty Nicovich wrote ----

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We have a project that involves imaging tissue sections mounted in
home-made Mowiol (PVA) mounting medium.  The coverslip is held away from
the slide with an adhesive spacer, the aperture in the spacer filled with
tissue and medium, and a coverslip applied.

These work great for about a month.  After that, we notice more and more
often bubbles appearing in the mounting medium. These are mostly around the
edge and immediately under the coverslip.

We've tried sealing with nail polish.  This seems to have little effect on
the rate or the extent of bubble formation.

My theory is that the PVA is shrinking and pulling away from the coverslip
and spacer.  If that's the case (or if not) is there a way to prevent
this?  Would another sealing material do the job better? Any tips or tricks
out there?

Thanks,
Rusty Nicovich

[daj1u06]

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if your tape chamber is a complete seal around the sample, surely the mowiol won't actually cure?

For sealing thin chambers we use this stuff, in the 25mm wide variety:
http://www.stix2.co.uk/double-sided-tapes/polyester-tapes
available online
and use a wad punch to punch out 15ish mm circular holes. It does what it says on the can, once it is on it will be virtually impossible to get off, so you need to sacrifice half of it to use its backing tape to cover the 2nd sticky side of a strip before punching the holes. We have samples cleared in TDE and the such like that have been sealed in it for several years now.

Regards,

Dave Johnston,
Biomedical Imaging Unit
Southampton, UK.