clearing_methods
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clearing_methods [2013/10/23 11:04] kthorn |
clearing_methods [2016/01/11 09:50] (current) kthorn |
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| 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 {{:mounting_media_tests.pdf|white paper}}. Since then, other clearing methods have been published: | 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 {{:mounting_media_tests.pdf|white paper}}. Since then, other clearing methods have been published: | ||
| + | * [[http://www.sciencedirect.com/science/article/pii/S0092867414009313|Passive Clarity (PACT)]] clearing has been used successfully by a number of UCSF labs. | ||
| * The [[http://www.ncbi.nlm.nih.gov/pubmed/21878933|Scale protocol]] has been published by the Miyawaki lab, which uses 4M urea to render whole mouse embryos essentially transparent. | * The [[http://www.ncbi.nlm.nih.gov/pubmed/21878933|Scale protocol]] has been published by the Miyawaki lab, which uses 4M urea to render whole mouse embryos essentially transparent. | ||
| * A [[http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0033916|recent paper]] shows that dehydration with tetrahydrofuran and clearing with dibenzyl ether preserves GFP fluorescence better than ethanol dehydration and clearing with BABB. | * A [[http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0033916|recent paper]] shows that dehydration with tetrahydrofuran and clearing with dibenzyl ether preserves GFP fluorescence better than ethanol dehydration and clearing with BABB. | ||
| * [[http://www.nature.com/nature/journal/v497/n7449/full/nature12107.html|CLARITY]] uses electrophoresis with SDS to clear very thick brain tissue. | * [[http://www.nature.com/nature/journal/v497/n7449/full/nature12107.html|CLARITY]] uses electrophoresis with SDS to clear very thick brain tissue. | ||
| + | * [[http://www.nature.com/neuro/journal/v16/n8/full/nn.3447.html|SeeDB]] uses fructose and thioglycerol to clear tissue and appears to work about as well as Scale. | ||
| * Here is a [[http://cshprotocols.cshlp.org/content/2012/9/pdb.prot069625.full|detailed protocol]] for fixing and mounting dragonfly tissue in 2,2-TDE. | * Here is a [[http://cshprotocols.cshlp.org/content/2012/9/pdb.prot069625.full|detailed protocol]] for fixing and mounting dragonfly tissue in 2,2-TDE. | ||
| + | * [[http://www.nature.com/nprot/journal/v7/n11/full/nprot.2012.119.html|3DISCO]] uses tissue dehydration in THF and clearing with dibenzyl ether. It clears quickly and is at least somewhat compatible with GFP. | ||
| * [[http://www.citifluor.com/non_hardening_antifadents.php|Citifluor]] mounting media are supposed to clear well (Confocal Listserv post from Sean Speese, 10/22/2013). | * [[http://www.citifluor.com/non_hardening_antifadents.php|Citifluor]] mounting media are supposed to clear well (Confocal Listserv post from Sean Speese, 10/22/2013). | ||
| + | * A [[http://onlinelibrary.wiley.com/doi/10.1002/anie.201306039/full|recent review]] compares a variety of these clearing methods. | ||
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