Clearing solution for thick pancreas sections

[Silke]

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Dear all,

For my future experiments, I am going to work with formalin-sucrose fixed human
cryotissue (pancreas). I would like to make slides of 400-500 micrometer.
After immunofluorescence staining and before using confocal microscopy, it will be
necessary to "clear" the tissue.

I have never done this before. Does someone have some recommendations for the
clearing solution that I should use?

Thanks!

Kind regards,
Silke

[Kurt Thorn]

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Hi Silke -

I've compiled a list of most of the recently published clearing methods
here: http://nic.ucsf.edu/dokuwiki/doku.php?id=clearing_methods

I don't think there is a clear winner yet; users of our facility have
reported widely varying results for different combinations of clearing
method and tissue type.

Kurt

On 12/11/2013 7:11 AM, Silke wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Dear all,
>
> For my future experiments, I am going to work with formalin-sucrose fixed human
> cryotissue (pancreas). I would like to make slides of 400-500 micrometer.
> After immunofluorescence staining and before using confocal microscopy, it will be
> necessary to "clear" the tissue.
>
> I have never done this before. Does someone have some recommendations for the
> clearing solution that I should use?
>
> Thanks!
>
> Kind regards,
> Silke
>
>

--
Kurt Thorn
Director, Nikon Imaging Center
http://nic.ucsf.edu/blog/

[Fassler, Matthias]

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Hi Silke,

at PerkinElmer we haven't been working with tissue sections but with tumor
microtissues that have a diameter of approx. 400 µm. We used the ScaleA2
reagent for optical clearing and we were really surprised how well it
actually worked. We also tried to quantify how deep we can image into the
microtissue before and after clearing. I'm not sure how our results
translate to your research, but in case your interested:
http://www.perkinelmer.com/pdfs/downloads/APP_Opera-Microtissue-Cores.pdf

The original ScaleA2 publication:
Hama, H., Kurokawa, H., Kawano, H., Ando, R., Shimogori, T., Noda, H.,
Fukami, K., Sakaue-Sawano, A., and Miyawaki, A. (2011): Scale: a chemical
approach for fluorescence imaging and reconstruction of transparent mouse
brain. Nature Neuroscience, 14 (11), 1481–8.

Best regards
Matthias Fassler
Application Scientist - HCS Applications
PerkinElmer, Hamburg