Rice Straw Tissue Preparation for Reproducible Electron Microscopy Imaging and Analysis

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.