Mucus Removal

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Ellen T. Arena Ellen T. Arena
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Mucus Removal

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I am hoping someone may have suggestions/protocols for effectively removing the mucus layer in intestinal tissue (especially a problem in colonic tissues) for whole-mount and/or live imaging.  I am having a great deal of trouble imaging the tissue with light-scattering of the mucus and difficulties removing the mucus without disturbing the tissue itself.  Does anyone have any suggestions?  It would be most helpful…

Many thanks.

Ellen



------------------------------------------------------
Ellen T. Arena, PhD
Pasteur Foundation Postdoctoral Fellow
Unité de Pathogénie Microbienne Moléculaire
INSERM U786
Institut Pasteur
28 rue du Dr Roux
F - 75724 PARIS Cédex 15
France
Tel: (33-0) 1 40 61 37 71
Fax: (33-0) 1 45 68 89 53
[hidden email]
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Ian Birchall Ian Birchall
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Re: Mucus Removal

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you could try a proteinase K digestion protocol

Ian Birchall
 Florey Institute
________________________________________
From: Confocal Microscopy List [[hidden email]] on behalf of Ellen T. Arena [[hidden email]]
Sent: Tuesday, 11 September 2012 7:22 PM
To: [hidden email]
Subject: Mucus Removal

*****
To join, leave or search the confocal microscopy listserv, go to:
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*****

I am hoping someone may have suggestions/protocols for effectively removing the mucus layer in intestinal tissue (especially a problem in colonic tissues) for whole-mount and/or live imaging.  I am having a great deal of trouble imaging the tissue with light-scattering of the mucus and difficulties removing the mucus without disturbing the tissue itself.  Does anyone have any suggestions?  It would be most helpful…

Many thanks.

Ellen



------------------------------------------------------
Ellen T. Arena, PhD
Pasteur Foundation Postdoctoral Fellow
Unité de Pathogénie Microbienne Moléculaire
INSERM U786
Institut Pasteur
28 rue du Dr Roux
F - 75724 PARIS Cédex 15
France
Tel: (33-0) 1 40 61 37 71
Fax: (33-0) 1 45 68 89 53
[hidden email]
------------------------------------------------------

Barbara Foster Barbara Foster
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Re: Mucus Removal

In reply to this post by Ellen T. Arena
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Hi, Ellen

Are you able to change the mounting material to
something like glycerin which might match the RI
of the mucus more closely but still be inert to
the whole mount?  I don't know that this will
work with living tissue, but it might be worth a
try with the whole mount.  We played a similar
trick on a materials science application when
trying to look at very fine copper particles on a filter.

Also, try minimizing the pinhole on your
confocal.  It should help reduce the scatter dramatically.

Good hunting!
Barbara Foster, President and Sr. Consultant

Microscopy/Microscopy Education
P: (972)924-5310
W: www.MicroscopyEducation.com

We are now scheduling courses through January 2013

At 10:27 PM 9/10/2012, Ellen T. Arena wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>I am hoping someone may have
>suggestions/protocols for effectively removing
>the mucus layer in intestinal tissue (especially
>a problem in colonic tissues) for whole-mount
>and/or live imaging.  I am having a great deal
>of trouble imaging the tissue with
>light-scattering of the mucus and difficulties
>removing the mucus without disturbing the tissue
>itself.  Does anyone have any suggestions?  It would be most helpful…
>
>Many thanks.
>
>Ellen
>
>
>
>------------------------------------------------------
>Ellen T. Arena, PhD
>Pasteur Foundation Postdoctoral Fellow
>Unité de Pathogénie Microbienne Moléculaire
>INSERM U786
>Institut Pasteur
>28 rue du Dr Roux
>F - 75724 PARIS Cédex 15
>France
>Tel: (33-0) 1 40 61 37 71
>Fax: (33-0) 1 45 68 89 53
>[hidden email]
>------------------------------------------------------