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Technical Note

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

1
Graduate School of Biological Sciences, Nara Institute of Science and Technology (NAIST), 8916-5 Takayama, Nara 630-0192, Japan
2
International Research Organization for Advanced Science and Technology (IROAST), Kumamoto University, 2-39-1 Kurokami, Chuo-ku, Kumamoto 860-8555, Japan
3
Faculty of Science, Kumamoto University, 2-39-1 Kurokami, Chuo-ku, Kumamoto 860-8555, Japan
4
Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Chuo-ku, Kumamoto 860-8555, Japan
*
Author to whom correspondence should be addressed.
Plants 2021, 10(2), 190; https://doi.org/10.3390/plants10020190
Received: 31 December 2020 / Revised: 17 January 2021 / Accepted: 18 January 2021 / Published: 20 January 2021
(This article belongs to the Special Issue Plant Embryogenesis)
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. View Full-Text
Keywords: clearing; 3D imaging; Arabidopsis thaliana; embryo; confocal microscopy; cell wall staining; fluorescent reporter; GFP clearing; 3D imaging; Arabidopsis thaliana; embryo; confocal microscopy; cell wall staining; fluorescent reporter; GFP
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MDPI and ACS Style

Imoto, A.; Yamada, M.; Sakamoto, T.; Okuyama, A.; Ishida, T.; Sawa, S.; Aida, M. A ClearSee-Based Clearing Protocol for 3D Visualization of Arabidopsis thaliana Embryos. Plants 2021, 10, 190. https://doi.org/10.3390/plants10020190

AMA Style

Imoto A, Yamada M, Sakamoto T, Okuyama A, Ishida T, Sawa S, Aida M. A ClearSee-Based Clearing Protocol for 3D Visualization of Arabidopsis thaliana Embryos. Plants. 2021; 10(2):190. https://doi.org/10.3390/plants10020190

Chicago/Turabian Style

Imoto, Ayame, Mizuki Yamada, Takumi Sakamoto, Airi Okuyama, Takashi Ishida, Shinichiro Sawa, and Mitsuhiro Aida. 2021. "A ClearSee-Based Clearing Protocol for 3D Visualization of Arabidopsis thaliana Embryos" Plants 10, no. 2: 190. https://doi.org/10.3390/plants10020190

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