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Quantitative analysis of SEM micrographs is essential to convert qualitative observations of starch granules into objective descriptors of size and shape. This chapter presents a reproducible SEM–ImageJ workflow that uses calibrated SEM images (JPG/TIF) to measure morphology and size distribution. The protocol covers scale calibration from the micrograph bar; preprocessing (8-bit conversion, contrast enhancement, light smoothing); segmentation (thresholding and watershed) to isolate touching granules; ROI curation; and measurement via Analyze Particles. Recommended outputs include area, perimeter, equivalent circular diameter (ECD), Feret max/min, aspect ratio, circularity, and solidity. We provide guidance on sampling (multiple fields; ≥200–300 granules), artifact control (edge objects, charging, compression), and standardized reporting of histograms, cumulative curves, and summary statistics. Metadata capture and macro saving are emphasized to ensure reproducibility and comparability across samples, instruments, and treatments.

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Morphology and Size Distribution of Starch Granules: A Sem–Imagej Quantitative Analysis Workflow

  • 章节

  • 发表日期：

    2026年

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  • Rebeca Salvador-Reyes,
  • Elisa Cristina Andrade Neves,
  • Roberto de Freitas Neves,
  • Maria Teresa Pedrosa Silva Clerici

摘要

Quantitative analysis of SEM micrographs is essential to convert qualitative observations of starch granules into objective descriptors of size and shape. This chapter presents a reproducible SEM–ImageJ workflow that uses calibrated SEM images (JPG/TIF) to measure morphology and size distribution. The protocol covers scale calibration from the micrograph bar; preprocessing (8-bit conversion, contrast enhancement, light smoothing); segmentation (thresholding and watershed) to isolate touching granules; ROI curation; and measurement via Analyze Particles. Recommended outputs include area, perimeter, equivalent circular diameter (ECD), Feret max/min, aspect ratio, circularity, and solidity. We provide guidance on sampling (multiple fields; ≥200–300 granules), artifact control (edge objects, charging, compression), and standardized reporting of histograms, cumulative curves, and summary statistics. Metadata capture and macro saving are emphasized to ensure reproducibility and comparability across samples, instruments, and treatments.