Transmission Electron Microscopy Imaging to Analyze Chromatin Density Distribution at the Nanoscale Level.

Abstract

Transmission electron microscopy (TEM) is used to study the fine ultrastructural organization of cells. Delicate specimen preparation is required for results to reflect the "native" ultrastructural organization of subcellular features such as the nucleus. Despite the advent of high-resolution, fluorescent imaging of chromatin components, TEM still provides a unique and complementary level of resolution capturing chromatin organization at the nanoscale level. Here, we describe the workflow, from tissue preparation, TEM image acquisition and image processing, for obtaining a quantitative description of chromatin density distribution in plant cells, informing on local fluctuations and periodicity. Comparative analyses then allow to elucidate the structural changes induced by developmental or environmental cues, or by mutations affecting specific chromatin modifiers at the nanoscale level. We argue that this approach remains affordable and merits a renewed interest by the plant chromatin community.

Plant phenotyping relevance

植物細胞のクロマチン密度分布を定量化するためのTEM撮像・画像処理ワークフロー自体が中心であり、発達・環境・変異による植物細胞状態の評価に再利用可能な手法を提示している。

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