Rice Straw Tissue Preparation for Reproducible Electron Microscopy Imaging and Analysis.

Abstract

Common problems in biological sample processing for scanning electron microscopy (SEM) include cell collapse and destruction. To overcome the challenges surrounding SEM micrograph preparation, dried rice stems were used to develop a specific set of protocols for processing dried plant samples. Dried stems are rehydrated with a glycerol solution and fixed in formalin-acetic-alcohol to avoid cell wall collapse or organ distortion. The protocols detailed here comprise the first published method for preparing SEM images of dried plant tissue. The protocols offer a cost-effective approach to obtaining high-quality micrographs, facilitating the reconstruction of growth processes and the study of plant cell wall features. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Pretreatment and preparation of rice straw samples at the heading stage Basic Protocol 2: Paraffin infiltration and embedding Basic Protocol 3: Preparation of microscopic sections Basic Protocol 4: Transferring, adhering, and expanding sections on slides Support Protocol: Preparation of gelatin slides before sectioning to affix samples Basic Protocol 5: Preparation of samples for SEM imaging Basic Protocol 6: SEM analysis Basic Protocol 7: Processing and analysis of SEM images using ImageJ software.

Plant phenotyping relevance

乾燥イネ組織のSEM画像取得・処理・解析プロトコル自体が中心で、植物細胞壁形態などの表現型観察を可能にする方法開発である。

Code and data availability

The article is a methods protocol for rice straw SEM preparation and ImageJ quantification. It contains no public phenotype dataset, image collection, analysis code, or model deposit; the Data Availability Statement explicitly states no data are shared beyond the article, and no supplement is present (pmc-prop-has-sup:

No evidence-backed public reproduction asset is currently recorded.