Fast and High-Resolution Imaging of Pollinated Stigmatic Cells by Tabletop Scanning Electron Microscopy.

Abstract

In plants, the first interaction between the pollen grain and the epidermal cells of the stigma is crucial for successful reproduction. When the pollen is accepted, it germinates, producing a tube that transports the two sperm cells to the ovules for fertilization. Confocal microscopy has been used to characterize the behavior of stigmatic cells post-pollination [1], but it is time-consuming since it requires the development of a range of fluorescent marker lines. Here, we propose a quick, high-resolution imaging protocol using tabletop scanning electron microscopy. This technique does not require prior sample fixation or fluorescent marker lines. It effectively captures pollen grain behavior from early hydration (a few minutes after pollination) to pollen tube growth within the stigma (1 h after pollination) and is particularly efficient for tracking pollen tube paths. Key features • Analysis of the pollen behavior in stigmatic cells of Arabidopsis thaliana but can be broadly used for other species. • Rapid and high-resolution imaging method. • Allows testing pollen grain hydration states, pollen tube paths on stigmatic cells from various genetic backgrounds, and also pollen tube phenotypes.

Plant phenotyping relevance

卓上走査電子顕微鏡を用いて花粉挙動、花粉管経路、花粉管表現型を取得する高速・高解像度イメージングプロトコルが研究の中心であり、植物表現型計測法の開発に該当する。

Code and data availability

This is a methods/protocol article describing tabletop SEM imaging of pollinated stigmas and ImageJ-based pollen tube turn quantification. No public phenotype/trait datasets, image collections, author analysis code, or trained models are deposited or linked; the only software mentioned is generic ImageJ, and data shown

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