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

Posted by Glen MacDonald-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/How-deep-can-you-see-in-a-brain-slice-tp7256661p7259377.html

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It partly depends on your resolution requirements.  George's response was characteristically thorough.

We've fixed whole mouse brain and cleared in 100% glycerol to record infiltration by GFP-expressing G6 glioma cell with 2 mm thick optical volume on a FV-1000 confocal, 4X and 10X/.4 objectives, as well as 400 um thick cultured slices.   Same instrument with 20X.75 objective could get >400 mm into 1 mm fixed mouse brain slices cleared with methyl salicylate-benzyl benzoate to create montages tracing axons (It was left over from imaging intact mouse cochlea at 800 um depth with 10X/.4, down to working distance of any high NA lens).   If using immersion lenses, don't overlook the benefits of adjusting the RI of your immersion medium.  

Regards,
Glen
Glen MacDonald
Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
Box 357923
University of Washington
Seattle, WA 98195-7923  USA
[phone]
[hidden email]








On Feb 5, 2012, at 11:53 AM, 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]