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http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear Colleagues, I am interested in learning about tissue sectioning equipment that is capable of accurately preserving the geometry of the slices (so successive sections can be aligned), and transferring the slices to slides. Any suggestions? Thanks! Badri Roysam Professor, Department of Electrical, Computer and Systems Engineering Associate Director, NSF Center for Subsurface Sensing & Imaging Systems (CenSSIS ERC) Rensselaer Polytechnic Institute 110 8th Street, Troy, New York 12180-3590. Office(JEC 7010): 518-276-8067, Lab(JEC 6308): 518-276-8207, Fax: 518-276-8715 Email: [hidden email], Web: http://www.ecse.rpi.edu/~roysam |
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal You might want to look into Instrumedics CryoJane Tape Transfer System: http://www.instrumedics.com/cryojane.htm It works for both frozen & paraffins. (I have no personal experience with this system but it may be useful for your application) Melissa Melissa A. González Edick R&D, Histology Cell Genesys Inc. 500 Forbes Blvd South San Francisco, CA 94080 p(650) 266-3168 f(650) 266-3080 -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Badri Roysam Sent: Wednesday, April 30, 2008 11:21 AM To: [hidden email] Subject: Tissue Sectioning Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear Colleagues, I am interested in learning about tissue sectioning equipment that is capable of accurately preserving the geometry of the slices (so successive sections can be aligned), and transferring the slices to slides. Any suggestions? Thanks! Badri Roysam Professor, Department of Electrical, Computer and Systems Engineering Associate Director, NSF Center for Subsurface Sensing & Imaging Systems (CenSSIS ERC) Rensselaer Polytechnic Institute 110 8th Street, Troy, New York 12180-3590. Office(JEC 7010): 518-276-8067, Lab(JEC 6308): 518-276-8207, Fax: 518-276-8715 Email: [hidden email], Web: http://www.ecse.rpi.edu/~roysam |
In reply to this post by Badri Roysam
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http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal On Wed, 30 Apr 2008, Badri Roysam wrote: > Search the CONFOCAL archive at > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > > Dear Colleagues, > > I am interested in learning about tissue sectioning equipment that is capable > of accurately preserving the geometry of the slices (so successive sections can > be aligned), and transferring the slices to slides. Any suggestions? > Are you talking about cutting from paraffin blocks? I hate to tell you this, but whatever you use probably won't work if you are going to try to do 3D reconstruction with multiple optical slices through each thick section. The knife introduces non-affine warping, and you will get non-affine deformation in the Z-axis as well, depending on the type of tissue. When we did this with glomeruli, the tufts would stick up out of the plane of the section, being pulled up by the knife. Since the initial alignment of sections is pretty simple in terms of rotation, translation, and shear, (and there are lots of tricks for automating it -- I used embedded fiducials) the hard part is the nonaffine deformation. Thus, I would concentrate most of my effort in getting a clean cut rather than worrying about getting a pre-aligned ribbon. As an aside, I would suggest looking at the ITK toolkit for developing registration software, if you haven't already. See: www.itk.org billo http://www.billoblog.com/billoblog |
In reply to this post by Badri Roysam
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Dear Badri,
I think the best way to achieve
what you want is microtomy on paraffin block. The structure will be perfect
through the entire section including the up and bottom faces. Cryostat
will provide you with successive sections but I would be surprised if you could
accurately align them due to the poor preservation of the structure after
freezing (especially on the faces of the section). Vibratome sectionning would
give you a perfect structure but the sections will show stripes on
their faces (but no more of 1 µm with a good apparatus). I don't know
the kind of resolution you need when you say accurately.
Bruno
Bruno SAUBAMEA
EA 3621 & Service Commun d'Imagerie Cellulaire et Moléculaire
Faculté des Sciences Pharmaceutiques et Biologiques
Université Paris Descartes 4, avenue de l'Observatoire 75006 PARIS tel : 01.53.73.97.13
fax : 01.53.73.99.09
|
In reply to this post by Badri Roysam
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi Bill, What type of technique gives the cleanest cut in your estimation? Badri Roysam Professor, Department of Electrical, Computer and Systems Engineering Associate Director, NSF Center for Subsurface Sensing & Imaging Systems (CenSSIS ERC) Rensselaer Polytechnic Institute 110 8th Street, Troy, New York 12180-3590. Office(JEC 7010): 518-276-8067, Lab(JEC 6308): 518-276-8207, Fax: 518-276-8715 Email: [hidden email], Web: http://www.ecse.rpi.edu/~roysam ----- Original Message ----- From: Bill Oliver [mailto:[hidden email]] To: [hidden email] Subject: Re: Tissue Sectioning > Search the CONFOCAL archive at > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > > On Wed, 30 Apr 2008, Badri Roysam wrote: > > > Search the CONFOCAL archive at > > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > > > > Dear Colleagues, > > > > I am interested in learning about tissue sectioning equipment that is > capable > > of accurately preserving the geometry of the slices (so successive > sections can > > be aligned), and transferring the slices to slides. Any suggestions? > > > > Are you talking about cutting from paraffin blocks? > > I hate to tell you this, but whatever you use probably won't work if you are > going to try to do 3D reconstruction with multiple optical slices through > each thick section. The knife introduces non-affine warping, and you will > get non-affine deformation in the Z-axis as well, depending on the type of > tissue. When we did this with glomeruli, the tufts would stick up out of > the plane of the section, being pulled up by the knife. > > Since the initial alignment of sections is pretty simple in terms of > rotation, translation, and shear, (and there are lots of tricks for > automating it -- I used embedded fiducials) the hard part is the nonaffine > deformation. Thus, I would concentrate most of my effort in getting a clean > cut rather than worrying about getting a pre-aligned ribbon. > > As an aside, I would suggest looking at the ITK toolkit for developing > registration software, if you haven't already. See: www.itk.org > > billo > http://www.billoblog.com/billoblog > |
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