3D Electron Microscopy of the ER.

Abstract

The endoplasmic reticulum (ER) forms an extensive network in plant cells. In leaf cells and vacuolated root cells it is mainly restricted to the cortex whereas in the root meristem the cortical and cytoplasmic ER takes up a large volume throughout the entire cell. Only 3D electron microscopy provides sufficient resolution to understand the spatial organization of the ER in the root. However, high contrast staining and optimally ER specific staining is essential. Here we describe a protocol for selective ER staining that allows automated or semiautomated segmentation of the organelle in 3D datasets obtained from serial sections, Array Tomography, Serial Block Face Scanning Electron Microscopy (SBFSEM), or Focused Ion Beam (FIB) SEM.

Plant phenotyping relevance

植物細胞内ERの3D画像取得・選択染色・自動／半自動セグメンテーションを可能にする手法プロトコルが中心であり、細胞内オルガネラの空間状態を抽出する植物フェノタイピング手法に該当する。

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