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.

中文翻译：

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基于ClearSee的清除协议，用于拟南芥胚胎的3D可视化

组织清除方法与共聚焦显微镜相结合已被广泛用于研究发育生物学。在植物中，ClearSee是一种可靠的清除方法，适用于广泛的组织，适用于使用荧光报告基因进行基因表达分析，但它尚未应用于拟南芥胚胎（用于研究形态发生和模式形成的模型系统）中在原始文献中已有描述。在这里，我们描述了一种基于ClearSee的清除协议，该协议适用于获得拟南芥的3D图像。胚胎。该方法包括胚胎解剖，固定，清洗，清除和细胞壁染色，并能够对胚胎形态进行高质量的3D成像，并以细胞分辨率表达荧光报告分子。我们的协议提供了一种可靠的方法，适用于拟南芥胚胎的形态发生和基因表达模式的分析。