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2020 Fiscal Year Final Research Report

Three-dimensional ultrastructural analysis of enteric glial cells with Cryo-fixation

Research Project

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Project/Area Number 17KK0191
Research Category

Fund for the Promotion of Joint International Research (Fostering Joint International Research)

Allocation TypeMulti-year Fund
Research Field Gastroenterology
Research InstitutionNagoya University

Principal Investigator

Tamada Hiromi  名古屋大学, 医学系研究科, 助教 (60712817)

Project Period (FY) 2018 – 2020
Keywords電子顕微鏡 / Cryo-fixation / 急速高圧固定 / スパイン / グリア細胞
Outline of Final Research Achievements

In this study in order to understand the native ultrastructure of enteric glial cells with novel three-dimensional electron microscopy, such as the Focused ion beam / Scanning electron microscopy (FIB/SEM), a snap freezing method of liquid nitrogen jets, combined with very high pressures (cryo-fixation), was analyzed. At first, with using the mouse cortex tissues, the difference among the cryo-fixation and conventional chemical-fixations was confirmed. As a result, dendritic spines showed significantly thinner necks with cryo-fixation. This work highlights the usefulness of cryo-fixation for obtaining a more native and detailed view of the architecture of cells and tissues.

Free Research Field

細胞組織学

Academic Significance and Societal Importance of the Research Achievements

新しい電子顕微鏡技術の発展に伴い、これまでは不可能であった電子顕微鏡像による定量的評価が可能となった。そのような中、従来の電顕用組織固定法である強いアルデヒドを用いた化学固定では、細胞膜の構造は明瞭に維持されるものの、細胞や組織本来の形態が消失する可能性が特に強く危惧される。本研究で検討した化学固定液を用いない急速高圧凍結固定法Cryo-fixationは、細胞や組織のより本来の姿に近い形態を維持するものと考えられ、今後のあらゆる生理機能の根幹をなす形態解析において重要な成果である。

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Published: 2022-01-27  

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