Re: [Histonet] Bubbles in Thick Sections - Suggestions?

From:Gayle Callis

Are your sections tape transferred?

I think you need the xylene or clearant to allow the flow of mounting media 
into and around all spaces of the thicker sections.  I would go back to 
using a permanent mounting media and mount the coverslip on sections just 
coming out of your clearant.  Less air is trapped if the solvent is there, 
although you can add bubbles, obviously.

Make sure the coverslip is lowered  onto a generous drop of mounting media 
so thicker section fills with media, do not overly squeege the excess from 
under coverslip -  you can even put the media on top of the section to help 
fill spaces in clearant wetted section.

If you coverslip air dried sections, thin out a permanent mounting media so 
it flows easily while coverslip is being mounted, you can also add a tiny 
drop of clearant to edge of mounted coverslip, it will flow under and join 
media, things fill in nicely.

Good luck

At 10:02 PM 8/12/2004, you wrote:
>We cut sections of tissue between 20 um and 30 um thick to view blood 
>vessels after being stained with CD31. We also section tissue at 
>thicknesses of 12 to 16 um for our H+E stains.
>We sometimes get bubbles when we coverslip with non-aqueous mounting media 
>and hate to see them in well-stained slides. Does anyone have any 
>suggestions for not getting bubbles in thick sections of tissue? After the 
>last xylene step we allow the sections to dry to ensure that the tissues 
>are not wet and we are currently using CureMount II, a non-aqueous 
>UV-cured mounting media from Instrumedics.
>Help me destroy the bubble monster! Please....
>Thanks in advance,
>Do you Yahoo!?
>Yahoo! Mail Address AutoComplete - You start. We finish.
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Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

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