Staining and Clearing of Arabidopsis Reproductive Tissue for Imaging of Fluorescent Proteins.

Abstract

Imaging of fluorescent proteins in whole-mount tissue is a powerful tool to understand growth and developmental processes, not only in plants. With the advent of genetically encoded fluorescent reporters, which specifically label reproductive cells in Arabidopsis, deep tissue imaging has become increasingly important for the study of plant reproduction. To penetrate the surrounding layers of maternal tissue, however, the tissue has to be cleared by homogenizing the refractive index of the sample, often leading to inactivation of fluorescent proteins. 2,2'-thiodiethanol (TDE) has recently been introduced as a clearing agent that allows the imaging of fluorescent proteins in a cleared plant tissue. Here, we describe a simple protocol that combines TDE-based tissue clearing with cell wall staining to outline cells that enable deep tissue imaging in reproductive structures of Arabidopsis thaliana.

Plant phenotyping relevance

Arabidopsis生殖組織の深部蛍光イメージングを可能にするTDE組織透明化・細胞壁染色プロトコルが中心であり、植物組織の形態・細胞状態の取得法に該当する。

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