Method for Combined Observation of Serial Sections of Stented Arteries Embedded in Resin by Light Microscopy and Transmission Electron Microscopy

Author:

Isobe Atsushi1,Iwatani Kouichi1,Souba Junko1,Terao Hisako1,Hagiwara Hitomi1,Kumagai Fumiaki2,Saito Yoshiaki2,Nagano Kasuke3,Tasaki Masako1

Affiliation:

1. Evaluation Center, R&D Administration and Promotion Department, Terumo Corporation, Kanagawa, Japan

2. Hatano Research Institute, Food and Drug Safety Center, Kanagawa, Japan

3. Nagano Toxicologic-Pathology Consulting, Kanagawa, Japan

Abstract

We have developed a new method for obtaining information on whole tissues by light microscopy (LM) and ultrastructural features by transmission electron microscopy (TEM). This method uses serial sections of a stented artery embedded in resin. Stents were implanted in porcine coronary arteries in this study. The heart was perfusion fixed in a 2% paraformaldehyde and 1.25% glutaraldehyde mixed solution. The stented artery was then removed, fixed in 1% osmium, embedded in Quetol 651 resin, and sectioned serially. For LM, the black color of osmium was removed from the section by immersion in periodic acid and hydrogen peroxide after deplasticization. These sections were stained with hematoxylin and eosin and Elastica–Masson trichrome stain. For TEM, thin sections were re-embedded in Quetol 812 resin by the resupinate method and cut into ultrathin sections. A clear, fine structure was obtained, and organelles, microvilli, and cell junctions in the endothelium were easily observed. The combined observation of adjacent specimens by LM and TEM enabled us to relate histopathological changes in the millimeter scale to those in the nanometer scale.

Publisher

SAGE Publications

Subject

Cell Biology,Toxicology,Molecular Biology,Pathology and Forensic Medicine

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