Clearing Methods
Imaging of thick (>50 um) tissue sections is often improved by clearing of the samples - treating them so that the normally opaque sample becomes clear. In 2007, we tested a number of clearing methods in conjunction with Nan Tang. The results are summarized in this white paper. Since then, other clearing methods have been published:
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The
Scale protocol has been published by the Miyawaki lab, which uses 4M urea to render whole mouse embryos essentially transparent.
A
recent paper shows that dehydration with tetrahydrofuran and clearing with dibenzyl ether preserves GFP fluorescence better than ethanol dehydration and clearing with BABB.
CLARITY uses electrophoresis with SDS to clear very thick brain tissue.
SeeDB uses fructose and thioglycerol to clear tissue and appears to work about as well as Scale.
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3DISCO uses tissue dehydration in THF and clearing with dibenzyl ether. It clears quickly and is at least somewhat compatible with GFP.
Citifluor mounting media are supposed to clear well (Confocal Listserv post from Sean Speese, 10/22/2013).
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