Re: How "deep" can you see in a brain slice?

Posted by Martin Wessendorf-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/How-deep-can-you-see-in-a-brain-slice-tp7256661p7259139.html

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Dear Dr. Busek--

What do you want to see?  The presence or absence of glia, or details on
the glial cells themselves?  If all you want to do is to establish the
presence of the glia, you may be able to get reasonably good results by
mounting the tissue between two coverslips and taking images from either
side of the "sandwich".  Gel-coat a coverslip, mount the tissue on it,
and then dehydrate, clear, and mount with another coverslip using DPX.
I'd image the tissue using oil-immersion optics, bearing in mind that
you want the longest working distance possible.  --Different
manufacturers' objectives have different capabilities in that regard and
you can't necessarily trust the specifications.

By mounting the tissue between two coverslips, you can get good optical
access to either side of the tissue.  You simply place the coverslip
"sandwich" on a slide and hold it in place with cellophane tape.  If you
don't need to create a single image through the entire thickness of the
tissue, it should work.

Good luck!

Martin Wessendorf

On 2/5/2012 1:53 PM, Petr Busek wrote:

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> Dear all,
> I am trying to view fluorescently labeled glioma cells invading into a 400um
> thick brain slice on an Olypus FV300. Has anyone experience with this and
> how "deep" it is reasonable to expect to see in the slice using a confocal
> microscope? How can you maximize this depth? (selection of objectives,
> processing of the slice....)
> Thanks for any suggestions, Petr.
>
> Petr Busek, MD, PhD
> Charles University in Prague
> First Faculty of Medicine
> Laboratory of Cancer Cell Biology
> Institute of Biochemistry and Experimental Oncology
> U Nemocnice 5
> 128 53 Prague 2
> Czech Republic
> www.lf1.cuni.cz/lbnb
>   Fax [phone]

--
Martin Wessendorf, Ph.D.                   office: [phone]
Assoc Prof, Dept Neuroscience                 lab: [phone]
University of Minnesota             Preferred FAX: [phone]
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