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A ClearSee-Based Clearing Protocol for 3D Visualization of Arabidopsis thaliana Embryos

Tissue clearing methods combined with confocal microscopy have been widely used for studying developmental biology. In plants, ClearSee is a reliable clearing method that is applicable to a wide range of tissues and is suitable for gene expression analysis using fluorescent reporters, but its applic...

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Detalles Bibliográficos
Autores principales: Imoto, Ayame, Yamada, Mizuki, Sakamoto, Takumi, Okuyama, Airi, Ishida, Takashi, Sawa, Shinichiro, Aida, Mitsuhiro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7909245/
https://www.ncbi.nlm.nih.gov/pubmed/33498275
http://dx.doi.org/10.3390/plants10020190
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author Imoto, Ayame
Yamada, Mizuki
Sakamoto, Takumi
Okuyama, Airi
Ishida, Takashi
Sawa, Shinichiro
Aida, Mitsuhiro
author_facet Imoto, Ayame
Yamada, Mizuki
Sakamoto, Takumi
Okuyama, Airi
Ishida, Takashi
Sawa, Shinichiro
Aida, Mitsuhiro
author_sort Imoto, Ayame
collection PubMed
description Tissue clearing methods combined with confocal microscopy have been widely used for studying developmental biology. In plants, ClearSee is a reliable clearing method that is applicable to a wide range of tissues and is suitable for gene expression analysis using fluorescent reporters, but its application to the Arabidopsis thaliana embryo, a model system to study morphogenesis and pattern formation, has not been described in the original literature. Here, we describe a ClearSee-based clearing protocol which is suitable for obtaining 3D images of Arabidopsis thaliana embryos. The method consists of embryo dissection, fixation, washing, clearing, and cell wall staining and enables high-quality 3D imaging of embryo morphology and expression of fluorescent reporters with the cellular resolution. Our protocol provides a reliable method that is applicable to the analysis of morphogenesis and gene expression patterns in Arabidopsis thaliana embryos.
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spelling pubmed-79092452021-02-27 A ClearSee-Based Clearing Protocol for 3D Visualization of Arabidopsis thaliana Embryos Imoto, Ayame Yamada, Mizuki Sakamoto, Takumi Okuyama, Airi Ishida, Takashi Sawa, Shinichiro Aida, Mitsuhiro Plants (Basel) Technical Note Tissue clearing methods combined with confocal microscopy have been widely used for studying developmental biology. In plants, ClearSee is a reliable clearing method that is applicable to a wide range of tissues and is suitable for gene expression analysis using fluorescent reporters, but its application to the Arabidopsis thaliana embryo, a model system to study morphogenesis and pattern formation, has not been described in the original literature. Here, we describe a ClearSee-based clearing protocol which is suitable for obtaining 3D images of Arabidopsis thaliana embryos. The method consists of embryo dissection, fixation, washing, clearing, and cell wall staining and enables high-quality 3D imaging of embryo morphology and expression of fluorescent reporters with the cellular resolution. Our protocol provides a reliable method that is applicable to the analysis of morphogenesis and gene expression patterns in Arabidopsis thaliana embryos. MDPI 2021-01-20 /pmc/articles/PMC7909245/ /pubmed/33498275 http://dx.doi.org/10.3390/plants10020190 Text en © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Technical Note
Imoto, Ayame
Yamada, Mizuki
Sakamoto, Takumi
Okuyama, Airi
Ishida, Takashi
Sawa, Shinichiro
Aida, Mitsuhiro
A ClearSee-Based Clearing Protocol for 3D Visualization of Arabidopsis thaliana Embryos
title A ClearSee-Based Clearing Protocol for 3D Visualization of Arabidopsis thaliana Embryos
title_full A ClearSee-Based Clearing Protocol for 3D Visualization of Arabidopsis thaliana Embryos
title_fullStr A ClearSee-Based Clearing Protocol for 3D Visualization of Arabidopsis thaliana Embryos
title_full_unstemmed A ClearSee-Based Clearing Protocol for 3D Visualization of Arabidopsis thaliana Embryos
title_short A ClearSee-Based Clearing Protocol for 3D Visualization of Arabidopsis thaliana Embryos
title_sort clearsee-based clearing protocol for 3d visualization of arabidopsis thaliana embryos
topic Technical Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7909245/
https://www.ncbi.nlm.nih.gov/pubmed/33498275
http://dx.doi.org/10.3390/plants10020190
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