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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3316521/ https://www.ncbi.nlm.nih.gov/pubmed/22479475 http://dx.doi.org/10.1371/journal.pone.0033916 |
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author | Becker, Klaus Jährling, Nina Saghafi, Saiedeh Weiler, Reto Dodt, Hans-Ulrich |
author_facet | Becker, Klaus Jährling, Nina Saghafi, Saiedeh Weiler, Reto Dodt, Hans-Ulrich |
author_sort | Becker, Klaus |
collection | PubMed |
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 | Online Article Text |
id | pubmed-3316521 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-33165212012-04-04 Chemical Clearing and Dehydration of GFP Expressing Mouse Brains Becker, Klaus Jährling, Nina Saghafi, Saiedeh Weiler, Reto Dodt, Hans-Ulrich PLoS One Research Article 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. Public Library of Science 2012-03-30 /pmc/articles/PMC3316521/ /pubmed/22479475 http://dx.doi.org/10.1371/journal.pone.0033916 Text en Becker et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Becker, Klaus Jährling, Nina Saghafi, Saiedeh Weiler, Reto Dodt, Hans-Ulrich Chemical Clearing and Dehydration of GFP Expressing Mouse Brains |
title | 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_short | Chemical Clearing and Dehydration of GFP Expressing Mouse Brains |
title_sort | chemical clearing and dehydration of gfp expressing mouse brains |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3316521/ https://www.ncbi.nlm.nih.gov/pubmed/22479475 http://dx.doi.org/10.1371/journal.pone.0033916 |
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