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 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.
['Imoto|Ayame|A|', 'Yamada|Mizuki|M|', 'Sakamoto|Takumi|T|', 'Okuyama|Airi|A|', 'Ishida|Takashi|T|0000-0001-5167-9971', 'Sawa|Shinichiro|S|0000-0002-9309-9104', 'Aida|Mitsuhiro|M|0000-0001-9000-7502']
clearing Arabidopsis thaliana embryo confocal microscopy fluorescent reporter 3D imaging cell wall staining GFP