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

Posted by Claudia Florindo on
URL: http://confocal-microscopy-list.275.s1.nabble.com/How-deep-can-you-see-in-a-brain-slice-tp7256661p7261717.html

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Dear Hugh

I have a user on my microscopy facility that has been using a combination of
2 protocols (using the scale reagent for making the samples transparent) and
a very strong permeabilization in order to make the antibody goo deep inside
the tissue.

The papers that he used where:

        Nature Neuroscience --> v14n11pg1482

        Microscopy and analysis --> Jan2011 pag5

I hope it helps

Good lick
Claudia.

______________________________________________

Claudia Florindo, PhD,  Microscopy Unit Manager
[hidden email]  /00351 289 244 489 ext:7489
Dept. de Ciências Biomédicas. e Medicina,
Uni. Algarve, Campus de Gambelas Ed. 8,  Lab 1.18
8005-139 Faro, Portugal



-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Hugh Newman
Sent: domingo, 5 de Fevereiro de 2012 20:14
To: [hidden email]
Subject: Re: How "deep" can you see in a brain slice?

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I'm no medical guy but I work for Japanese microscope company that will not
be named.
Brain isn't very translucent so I wouldn't hold out much hope. I would say
100um tops, maybe even closer to 50um. After this your image will start to
degrade quite a bit. Any deeper than that and you are going to want a
2P-Confocal. I've imaged colloidal suspensions that are ~100um thick, and
they are very close to being index matched. At the maximum depth the image
wasn't that pretty.

Maybe some more Neuro people will have a better answer for you.

Good luck

H

Hugh Newman
 
Graduate Researcher

Dept. Physics and Physical Oceanography

Memorial University

St. Johns, Newfoundland, Canada

> Date: Sun, 5 Feb 2012 13:53:33 -0600
> From: [hidden email]
> Subject: How "deep" can you see in a brain slice?
> To: [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
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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]