Chemical clearing and dehydration of GFP expressing mouse brains.

Generally, chemical tissue clearing is performed by a solution consisting of two parts benzyl benzoate and one part benzyl alcohol. However, prolonged exposure to this mixture markedly reduces the fluorescence of GFP expressing specimens, so that one has to compromise between clearing quality and fl...

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Autores principales:	Klaus Becker, Nina Jährling, Saiedeh Saghafi, Reto Weiler, Hans-Ulrich Dodt
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/c8a46938fac3465189af428831afe271
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spelling	oai:doaj.org-article:c8a46938fac3465189af428831afe2712021-11-18T07:23:40ZChemical clearing and dehydration of GFP expressing mouse brains.1932-620310.1371/journal.pone.0033916https://doaj.org/article/c8a46938fac3465189af428831afe2712012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22479475/?tool=EBIhttps://doaj.org/toc/1932-6203Generally, chemical tissue clearing is performed by a solution consisting of two parts benzyl benzoate and one part benzyl alcohol. However, prolonged exposure to this mixture markedly reduces the fluorescence of GFP expressing specimens, so that one has to compromise between clearing quality and fluorescence preservation. This can be a severe drawback when working with specimens exhibiting low GFP expression rates. Thus, we screened for a substitute and found that dibenzyl ether (phenylmethoxymethylbenzene, CAS 103-50-4) can be applied as a more GFP-friendly clearing medium. Clearing with dibenzyl ether provides improved tissue transparency and strikingly improved fluorescence intensity in GFP expressing mouse brains and other samples as mouse spinal cords, or embryos. Chemical clearing, staining, and embedding of biological samples mostly requires careful foregoing tissue dehydration. The commonly applied tissue dehydration medium is ethanol, which also can markedly impair GFP fluorescence. Screening for a substitute also for ethanol we found that tetrahydrofuran (CAS 109-99-9) is a more GFP-friendly dehydration medium than ethanol, providing better tissue transparency obtained by successive clearing. Combined, tetrahydrofuran and dibenzyl ether allow dehydration and chemical clearing of even delicate samples for UM, confocal microscopy, and other microscopy techniques.Klaus BeckerNina JährlingSaiedeh SaghafiReto WeilerHans-Ulrich DodtPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 3, p e33916 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Klaus Becker Nina Jährling Saiedeh Saghafi Reto Weiler Hans-Ulrich Dodt Chemical clearing and dehydration of GFP expressing mouse brains.
description	Generally, chemical tissue clearing is performed by a solution consisting of two parts benzyl benzoate and one part benzyl alcohol. However, prolonged exposure to this mixture markedly reduces the fluorescence of GFP expressing specimens, so that one has to compromise between clearing quality and fluorescence preservation. This can be a severe drawback when working with specimens exhibiting low GFP expression rates. Thus, we screened for a substitute and found that dibenzyl ether (phenylmethoxymethylbenzene, CAS 103-50-4) can be applied as a more GFP-friendly clearing medium. Clearing with dibenzyl ether provides improved tissue transparency and strikingly improved fluorescence intensity in GFP expressing mouse brains and other samples as mouse spinal cords, or embryos. Chemical clearing, staining, and embedding of biological samples mostly requires careful foregoing tissue dehydration. The commonly applied tissue dehydration medium is ethanol, which also can markedly impair GFP fluorescence. Screening for a substitute also for ethanol we found that tetrahydrofuran (CAS 109-99-9) is a more GFP-friendly dehydration medium than ethanol, providing better tissue transparency obtained by successive clearing. Combined, tetrahydrofuran and dibenzyl ether allow dehydration and chemical clearing of even delicate samples for UM, confocal microscopy, and other microscopy techniques.
format	article
author	Klaus Becker Nina Jährling Saiedeh Saghafi Reto Weiler Hans-Ulrich Dodt
author_facet	Klaus Becker Nina Jährling Saiedeh Saghafi Reto Weiler Hans-Ulrich Dodt
author_sort	Klaus Becker
title	Chemical clearing and dehydration of GFP expressing mouse brains.
title_short	Chemical clearing and dehydration of GFP expressing mouse brains.
title_full	Chemical clearing and dehydration of GFP expressing mouse brains.
title_fullStr	Chemical clearing and dehydration of GFP expressing mouse brains.
title_full_unstemmed	Chemical clearing and dehydration of GFP expressing mouse brains.
title_sort	chemical clearing and dehydration of gfp expressing mouse brains.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/c8a46938fac3465189af428831afe271
work_keys_str_mv	AT klausbecker chemicalclearinganddehydrationofgfpexpressingmousebrains AT ninajahrling chemicalclearinganddehydrationofgfpexpressingmousebrains AT saiedehsaghafi chemicalclearinganddehydrationofgfpexpressingmousebrains AT retoweiler chemicalclearinganddehydrationofgfpexpressingmousebrains AT hansulrichdodt chemicalclearinganddehydrationofgfpexpressingmousebrains
_version_	1718423562812915712

Chemical clearing and dehydration of GFP expressing mouse brains.

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