Tissue Sectioning

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Badri Roysam Badri Roysam
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Tissue Sectioning

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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
Melissa Gonzalez Melissa Gonzalez
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Re: Tissue Sectioning

Search the CONFOCAL archive at
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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
Bill Oliver-3 Bill Oliver-3
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Re: Tissue Sectioning

In reply to this post by Badri Roysam
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
Bruno Saubaméa Bruno Saubaméa
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Re: Tissue Sectioning

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
----- Original Message -----
Sent: Wednesday, April 30, 2008 8:21 PM
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
Badri Roysam Badri Roysam
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Re: Tissue Sectioning

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
>