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Improved Serial Sectioning Techniques for Correlative Light-Electron Microscopy Mapping of Human Langerhans Islets

Sei Saitoh1,2, Nobuhiko Ohno1, Yurika Saitoh1

  • 1Department of Anatomy and Molecular Histology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, 1110 Shimokato, Chuo, Yamanashi 409-3898, Japan.

Acta Histochemica Et Cytochemica
|April 7, 2018
PubMed

Insights

This study presents a new correlative microscopy method for analyzing human pancreatic islet cells. It combines immunofluorescence and electron microscopy to visualize both cellular structures and hormone localization in single cells.

Area of Science:

  • Endocrinology
  • Cell Biology
  • Microscopy

Background:

  • Combined immunostaining and ultrastructural analysis are vital for studying cell function.
  • Conventional fixation methods yield weak antigenicity, hindering immunoassays.
  • Human pancreatic islets require advanced imaging for detailed cellular analysis.

Purpose of the Study:

  • To develop a correlative light and electron microscopy (CLEM) method for human pancreatic islet cells.
  • To overcome antigenicity issues in fixed tissues for combined imaging.
  • To visualize ultrastructural features alongside immunolocalization of hormones and REG1α in the same cells.

Main Methods:

  • Utilized serial sections from Epon-embedded human pancreatic tissue.
  • Applied double-immunofluorescence for insulin, glucagon, and REG1α.
  • Removed epoxy resin and treated with hydrogen peroxide for de-osmification.
  • Superimposed immunofluorescence images with serial ultrathin section electron microscopy images.

Main Results:

  • Successfully visualized endocrine hormones (insulin, glucagon) and REG1α via immunofluorescence.
  • Electron microscopy revealed detailed ultrastructural features, including secretory granules and organelles.
  • Demonstrated successful superimposition of light and electron microscopy images of the same cells.

Conclusions:

  • The developed CLEM approach enables simultaneous examination of ultrastructure and immunolocalization in human pancreatic islets.
  • This method enhances the study of cellular functions in both normal and pathological pancreatic islet conditions.
  • The technique provides a powerful tool for detailed analysis of endocrine cell biology.

Keywords:
Epon-embeddedcompact and diffuse isletscorrelative microscopyhormoneimmunostaining

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