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New CLARITY system


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#1 Ahrenhase

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Posted 30 August 2013 - 03:13 PM

Has anyone tried this and had success at this?  I was interested in trying this for some brain samples. 



#2 miST32

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Posted 08 February 2014 - 10:23 AM

Yes.  I've tried it and had mixed/poor results.

If you follow the protocol provided on the clarity website, you'll probably get a clear brain.  However, you'll need to spend time optimizing the osmolarity of the solutions you use.  I've noticed that transferring the sample from clearing solution to imaging solution induces a rapid condensation of the tissue, which will destroy any morphology that you see under the microscope.  This is particularly troublesome if you're using a glycerol solution for imaging.

That said, I was able to identify cell bodies and projections on a Thy1-YFP mouse using a high NA objective and confocal system.  We captured a neat ~600 micron thick section and the 3D reconstruction of the z-sections turned out fairly well.  I hope to get back to it, it has potential.

I'd suggest trying it on small sections first and scaling up to the whole brain later, to optimize/minimize the osmotic damage to your tissue.

Cheers and good luck.


 






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