Improved serial sectioning techniques for correlative light-electron microscopy mapping of human langerhans islets

Sei Saitoh, Nobuhiko Ohno, Yurika Saitoh, Nobuo Terada, Satoshi Shimo, Kaoru Aida, Hideki Fujii, Tetsuro Kobayashi, Shinichi Ohno

研究成果: Article査読

3 被引用数 (Scopus)

抄録

Combined analysis of immunostaining for various biological molecules coupled with investigations of ultrastructural features of individual cells is a powerful approach for studies of cellular functions in normal and pathological conditions. However, weak antigenicity of tissues fixed by conventional methods poses a problem for immunoassays. This study introduces a method of correlative light and electron microscopy imaging of the same endocrine cells of compact and diffuse islets from human pancreatic tissue specimens. The method utilizes serial sections obtained from Epon-embedded specimens fixed with glutaraldehyde and osmium tetroxide. Double-immunofluorescence staining of thick Epon sections for endocrine hormones (insulin and glucagon) and regenerating islet-derived gene 1 α (REG1α) was performed following the removal of Epoxy resin with sodium ethoxide, antigen retrieval by autoclaving, and de-osmification treatment with hydrogen peroxide. The immunofluorescence images of endocrine cells were superimposed with the electron microscopy images of the same cells obtained from serial ultrathin sections. Immunofluorescence images showed well-preserved secretory granules in endocrine cells, whereas electron microscopy observations demonstrated corresponding secretory granules and intracellular organelles in the same cells. In conclusion, the correlative imaging approach developed by us may be useful for examining ultrastructural features in combination with immunolocalisation of endocrine hormones in the same human pancreatic islets.

本文言語English
ページ(範囲)9-20
ページ数12
ジャーナルActa Histochemica et Cytochemica
51
1
DOI
出版ステータスPublished - 2018
外部発表はい

All Science Journal Classification (ASJC) codes

  • Pathology and Forensic Medicine
  • Biochemistry
  • Physiology
  • Histology
  • Cell Biology

フィンガープリント 「Improved serial sectioning techniques for correlative light-electron microscopy mapping of human langerhans islets」の研究トピックを掘り下げます。これらがまとまってユニークなフィンガープリントを構成します。

引用スタイル