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Development of a new dielectric microscope and analysis of the linkage mechanism of intracellular organelles

Research Project

Project/Area Number 19H03230
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 44010:Cell biology-related
Research InstitutionNational Institute of Advanced Industrial Science and Technology

Principal Investigator

Ogura Toshihiko  国立研究開発法人産業技術総合研究所, 生命工学領域, 上級主任研究員 (70371028)

Co-Investigator(Kenkyū-buntansha) 山本 条太郎  国立研究開発法人産業技術総合研究所, 生命工学領域, 研究員 (20585088)
岡田 知子  国立研究開発法人産業技術総合研究所, 生命工学領域, 総括研究主幹 (30344146)
Project Period (FY) 2019-04-01 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥17,160,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥3,960,000)
Fiscal Year 2022: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2021: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2019: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000)
Keywords窒化シリコン薄膜 / 走査電子顕微鏡 / 誘電率 / 液中観察 / 培養細胞 / 細胞内小器官 / 走査電子誘電率顕微鏡 / 画像処理 / 3次元構造解析 / オートファジー / 走査電子 / 3次元観察
Outline of Research at the Start

細胞は、核や小胞体、ゴルジ体等の様々な細胞内小器官が相互に関連し連携することで生命活動を維持し様々な機能を発現している。本研究では、これまで観察が極めて困難であった生きた無処理の細胞内小器官を高分解能でかつその3次元構造をそのまま観察する新たな誘電率顕微鏡の開発を目指す。我々はこれまで電子線を用いた世界初の走査電子誘電率顕微鏡の開発を進めてきた。本提案では、この誘電率顕微鏡を大幅に高機能化し、さらに3次元構造を解析できる新たなシステムを開発する。加えて、本提案により開発した3次元観察システムを用いて、膜タンパク質複合体やオートファジ―等に関連する細胞内小器官の3次元構造変化を詳細に分析する。

Outline of Final Research Achievements

In this research, we improved the functionality of the scanning electron dielectric microscope. To achieve this, we developed a high-sensitivity, low-noise first-stage amplifier and a high-resolution observation holder with a 10-nm-thick SiN thin film. By increasing the thickness of the silicon nitride thin film to 10 nm, the spatial resolution was improved to 4.5 nm, making it possible to observe the structure of cells and organic materials in greater detail. By using this system, it is possible to directly observe various cells such as mouse breast cancer cells, melanin pigment cells, and oral epithelial cells at the nano level. Furthermore, from the observed images, we succeeded in analyzing the movement mechanism of intracellular organelles such as the intracellular membrane. In addition, by using the latest image information processing technology, we have developed an automatic analysis system for intracellular vesicles.

Academic Significance and Societal Importance of the Research Achievements

本研究では、高機能な走査電子誘電率顕微鏡を開発した。本システムを用いる事で、培養細胞の内部構造をナノレベルの分解能で生きたまま直接観察する事が可能となった。これにより、細胞内小器官のより詳細な解析が進展する事が予想される。さらに、培養細胞だけでなく、バクテリアやタンパク質複合体等の様々な生物試料の溶液中での直接観察と分析を可能とする。これ以外にも溶液中の有機材料やナノ粒子、食品や化粧品、石油化学製品、燃料電池や2次電池材料等の様々な分野においても活用が期待され、学術的にも産業的にも極めて大きな意義があるものと考えられる。

Report

(5 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Annual Research Report
  • 2020 Annual Research Report
  • 2019 Annual Research Report
  • Research Products

    (21 results)

All 2023 2022 2021 2020 2019

All Journal Article (11 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 11 results,  Open Access: 10 results) Presentation (10 results) (of which Int'l Joint Research: 5 results,  Invited: 9 results)

  • [Journal Article] Structural analysis of melanosomes in living mammalian cells using scanning electron-assisted dielectric microscopy with deep neural network2023

    • Author(s)
      Tomoko Okada, Tomoaki Iwayama, Taku Ogura, Shinya Murakami, Toshihiko Ogura
    • Journal Title

      Computational and Structural Biotechnology Journal

      Volume: 21 Pages: 506-518

    • DOI

      10.1016/j.csbj.2022.12.027

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Development of General-purpose Dielectric Constant Imaging Unit for SEM and Direct Observation of Samples in Aqueous Solution2023

    • Author(s)
      Toshihiko Ogura, Tomoko Okada, Michio Hatano, Mitsuhiro Nakamura, Toshihide Agemura
    • Journal Title

      Microscopy and Microanalysis

      Volume: - Issue: 3 Pages: 1037-1046

    • DOI

      10.1093/micmic/ozad030

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Giant Carbon Nano-Test Tubes as Versatile Imaging Vessels for High-Resolution and In Situ Observation of Proteins2022

    • Author(s)
      Tracy T Chuong, Toshihiko Ogura, Norihito Hiyoshi, Kazuma Takahashi, Sangho Lee, Keita Hiraga, Hiroki Iwase, Akira Yamaguchi, Kiyoto Kamagata, Eriko Mano, Satoshi Hamakawa, Hirotomo Nishihara, Takashi Kyotani, Galen D Stucky, Tetsuji Itoh
    • Journal Title

      ACS Applied Materials & Interfaces

      Volume: 14 Issue: 23 Pages: 26507-26516

    • DOI

      10.1021/acsami.2c06318

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Arginine and its Derivatives Suppress the Opalescence of an Antibody Solution2022

    • Author(s)
      Shogo Oki, Suguru Nishinami, Yoshitaka Nakauchi, Toshihiko Ogura, Kentaro Shiraki
    • Journal Title

      Journal of Pharmaceutical Sciences

      Volume: 111 Issue: 4 Pages: 1126-1132

    • DOI

      10.1016/j.xphs.2021.11.018

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Development of multi-frequency impedance scanning electron microscopy2022

    • Author(s)
      Ogura Toshihiko
    • Journal Title

      PLOS ONE

      Volume: 17 Issue: 1 Pages: e0263098-e0263098

    • DOI

      10.1371/journal.pone.0263098

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Raman scattering enhancement of dielectric microspheres on silicon nitride film2022

    • Author(s)
      Ogura Toshihiko
    • Journal Title

      Scientific Reports

      Volume: 12 Issue: 1

    • DOI

      10.1038/s41598-022-09315-5

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Nanoscale observation of PM2.5 incorporated into mammalian cells using scanning electron-assisted dielectric microscope2021

    • Author(s)
      Okada Tomoko、Iwayama Tomoaki、Murakami Shinya、Torimura Masaki、Ogura Toshihiko
    • Journal Title

      Scientific Reports

      Volume: 11 Issue: 1 Pages: 228-228

    • DOI

      10.1038/s41598-020-80546-0

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Scanning Electron-Assisted Dielectric Microscopy Reveals Autophagosome Formation by LC3 and ATG12 in Cultured Mammalian Cells2021

    • Author(s)
      Okada Tomoko、Ogura Toshihiko
    • Journal Title

      International Journal of Molecular Sciences

      Volume: 22 Issue: 4 Pages: 1834-1834

    • DOI

      10.3390/ijms22041834

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Direct observation of unstained biological samples in water using newly developed impedance scanning electron microscopy2019

    • Author(s)
      Ogura Toshihiko
    • Journal Title

      PLOS ONE

      Volume: 14 Issue: 8 Pages: e0221296-e0221296

    • DOI

      10.1371/journal.pone.0221296

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Development of INSOL‐tag for proteome‐wide protein handling and its application in protein array analysis2019

    • Author(s)
      Fukuda Eriko、Mori Masatoshi、Shiku Hiroshi、Miyahara Yoshihiro、Kawamura Yoshifumi、Ogawa Koji、Ogura Toshihiko、Goshima Naoki
    • Journal Title

      Genes to Cells

      Volume: 25 Issue: 1 Pages: 41-53

    • DOI

      10.1111/gtc.12735

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Osteoblastic lysosome plays a central role in mineralization2019

    • Author(s)
      Iwayama Tomoaki、Okada Tomoko、Ueda Tsugumi、Tomita Kiwako、Matsumoto Shuji、Takedachi Masahide、Wakisaka Satoshi、Noda Takeshi、Ogura Taku、Okano Tomomichi、Fratzl Peter、Ogura Toshihiko、Murakami Shinya
    • Journal Title

      Science Advances

      Volume: 5 Issue: 7

    • DOI

      10.1126/sciadv.aax0672

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] Nanoscale observation of fine particles and biological specimens in aqueous condition by scanning-elect ron assisted dielectric microscop2022

    • Author(s)
      小椋 俊彦、岡田 知子
    • Organizer
      11th International Conference on Fine Particle s Magnetis
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Nanoscale imaging of intact biological specimens in water using scanning electron assisted dielectric mi croscop2022

    • Author(s)
      小椋 俊彦、岡田 知子
    • Organizer
      第8回メタロミクスに関する国際会議(ISM-8)
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Direct observation of biological fine particles in water by scanning-electron assisted dielectric microscopy2021

    • Author(s)
      小椋俊彦、岡田知子
    • Organizer
      Pacifichem 2021
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 走査電子誘電率顕微鏡の開発と水溶液中の試料の直接観察2021

    • Author(s)
      小椋俊彦
    • Organizer
      第45回静電気学会全国大会
    • Related Report
      2021 Annual Research Report
    • Invited
  • [Presentation] 走査電子誘電率顕微鏡による溶液中のコロイド粒子や生物試料の直接観察2021

    • Author(s)
      小椋俊彦
    • Organizer
      コロイド先端 技術講座「先端バイオ計測技術」
    • Related Report
      2021 Annual Research Report
    • Invited
  • [Presentation] High-resolution imaging of proteins and cells under aqueous condition using scanning-electron assisted dielectric microscopy (SE-ADM)2020

    • Author(s)
      小椋 俊彦、伊藤 徹二、岡田 知子
    • Organizer
      第30回日本MRS年次大会
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] 革新的液中ナノ顕微鏡開発と細胞外微粒子の包括的解明2020

    • Author(s)
      小椋 俊彦
    • Organizer
      日本分析化学会第69回年会
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] Nanoscale observation of biological specimens in aqueous condition by scanning-electron assisted dielectric microscopy2019

    • Author(s)
      小椋 俊彦、岡田 知子
    • Organizer
      OKINAWA COLLOIDS 2019 conference
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Nanoscale imaging of unstained biological specimens in water using newly developed scanning-electron assisted dielectric microscopy2019

    • Author(s)
      小椋 俊彦、岡田 知子
    • Organizer
      5th International Kyushu Colloid Colloquium
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 走査電子誘電率顕微鏡によるナノ・マイクロプラスチックの細胞への影響解析2019

    • Author(s)
      小椋 俊彦、岡田 知子
    • Organizer
      産総研・シンポジウム「マイクロプラスチックの計測・評価を考える」
    • Related Report
      2019 Annual Research Report
    • Invited

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Published: 2019-04-18   Modified: 2024-01-30  

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