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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] ------------------------------------------------------ |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** 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: 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] ------------------------------------------------------ |
In reply to this post by Ellen T. Arena
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** 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] >------------------------------------------------------ |
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