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Development of nanoendoscopy techinique for direct visualization of nanodynamics inside living cells

Research Project

Project/Area Number 19K22125
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 29:Applied condensed matter physics and related fields
Research InstitutionKanazawa University

Principal Investigator

Fukuma Takeshi  金沢大学, ナノ生命科学研究所, 教授 (90452094)

Project Period (FY) 2019-06-28 – 2022-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2021: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2020: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2019: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Keywords原子間力顕微鏡(AFM) / 原子間力顕微鏡 / ナノ内視鏡 / 細胞内計測 / 細胞観察 / がん / 3次元AFM
Outline of Research at the Start

生きた細胞の内部にナノプローブを挿入し、内部の構造や動態をナノレベルで直接観察できる「ナノ内視鏡技術」を開発する。さらに、開発した技術を用いて、骨芽細胞が骨形成する過程において、細胞内で生じるナノ動態を直接可視化し、その生命科学分野における有用性を実証する。原子間力顕微鏡(AFM)は、液中で、非染色で、細胞やタンパク質のナノ動態を直接可視化できる唯一の計測技術である。しかし、従来のAFM技術では細胞内部のナノ動態までは観察できない。本研究によりナノ内視鏡技術が開発できれば、細胞内部におけるナノスケールの動態を直接見て理解することが可能となり、生命科学分野に大きな革新をもたらすことができる。

Outline of Final Research Achievements

We have developed "nanoendoscopy" that allows to visualize nanodynamics inside living cells based on atomic force microscopy. This technology visualizes intracellular structures by scanning a needle tip in three-dimensional space, including inside the cell, and recording the forces acting on the tip during the scanning. Furthermore, by inserting the needle into the cell and performing AFM observation there, the local intracellular structure and its dynamics can be observed with nano-level resolution. In addition, it was confirmed by fluorescence assay that these measurements do not cause lethal damage to the cells. This technology enables direct observation of intracellular phenomena that could not be observed previously, and is expected to advance our understanding of the mechanisms underlying the expression of cellular functions.

Academic Significance and Societal Importance of the Research Achievements

本研究の成果により、従来技術では観られなかった生細胞内部のナノ現象が観察できるようになる。これにより、従来間接的な計測結果から予想されてきた細胞内現象の発現機構やそれが関係する疾患の機序を直接観て確認できる。また、従来、細胞外での再構成系で多数のAFM計測が行われてきたが、それが本当に細胞内で生じているのかは、しばしば疑問視されてきた。これらの現象を直接細胞内で観察できれば、過去に報告されてきた膨大な量のAFM実験結果に関して、この疑問に対する解答を与えられる。こうして明らかになった細胞機能や疾患機序の理解は、将来、創薬・医学の発展を通じて、人類の健康増進に寄与することが期待される。

Report

(4 results)
  • 2021 Annual Research Report   Final Research Report ( PDF )
  • 2020 Research-status Report
  • 2019 Research-status Report
  • Research Products

    (31 results)

All 2022 2021 2020 2019 Other

All Journal Article (2 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 2 results,  Open Access: 2 results) Presentation (27 results) (of which Int'l Joint Research: 7 results,  Invited: 21 results) Remarks (2 results)

  • [Journal Article] Cell penetration efficiency analysis of different atomic force microscopy nanoneedles into living cells2021

    • Author(s)
      Penedo Marcos、Shirokawa Tetsuya、Alam Mohammad Shahidul、Miyazawa Keisuke、Ichikawa Takehiko、Okano Naoko、Furusho Hirotoshi、Nakamura Chikashi、Fukuma Takeshi
    • Journal Title

      Scientific Reports

      Volume: 11 Issue: 1 Pages: 7756-7756

    • DOI

      10.1038/s41598-021-87319-3

    • Related Report
      2021 Annual Research Report 2020 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Visualizing intracellular nanostructures of living cells by nanoendoscopy-AFM2021

    • Author(s)
      Penedo Marcos、Miyazawa Keisuke、Okano Naoko、Furusho Hirotoshi、Ichikawa Takehiko、Alam Mohammad Shahidul、Miyata Kazuki、Nakamura Chikashi、Fukuma Takeshi
    • Journal Title

      Science Advances

      Volume: 7 Issue: 52

    • DOI

      10.1126/sciadv.abj4990

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] AFMによる3次元自己組織化構造の内部観察2022

    • Author(s)
      福間剛士
    • Organizer
      ISSP Workshop「機能的走査プローブ顕微鏡の新展開」
    • Related Report
      2021 Annual Research Report
    • Invited
  • [Presentation] Visualizing Unexplored Nanoworld by Innovative In-liquid AFM Technologies2022

    • Author(s)
      Takeshi Fukuma
    • Organizer
      The 99th CEMS Colloquium
    • Related Report
      2021 Annual Research Report
    • Invited
  • [Presentation] Visualizing Intracellular Nanostructures of Living Cells by Nanoendoscopy-AFM2022

    • Author(s)
      Takeshi Fukuma
    • Organizer
      5th NanoLSI International Symposium
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 革新的液中原子間力顕微鏡技術による未踏ナノ領域の可視化2021

    • Author(s)
      福間剛士
    • Organizer
      ナノ科学シンポジウム2021
    • Related Report
      2021 Annual Research Report
    • Invited
  • [Presentation] 生細胞内部を可視化するナノ内視鏡の開発2021

    • Author(s)
      宮澤佳甫,福間剛士
    • Organizer
      2021年日本表面真空学会学術講演会
    • Related Report
      2021 Annual Research Report
    • Invited
  • [Presentation] Visualizing Inside of 3D Self-Organizing Systems by 3D-AFM2021

    • Author(s)
      Takeshi Fukuma
    • Organizer
      AVS 67 Virtual Symposium
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 金沢大学ナノ生命科学研究所における活動の紹介2021

    • Author(s)
      福間剛士
    • Organizer
      ナノプローブテクノロジー第167委員会第98回研究会
    • Related Report
      2021 Annual Research Report
    • Invited
  • [Presentation] Visualizing Inside of 3D Self-Organizing Systems by 3D-AFM2021

    • Author(s)
      Takeshi Fukuma
    • Organizer
      The 2021 I(SPM)3 Conference
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 革新的ナノプローブ技術によるナノ生命科学研究の新展開2021

    • Author(s)
      福間剛士
    • Organizer
      第73回日本細胞生物学会大会
    • Related Report
      2021 Annual Research Report
    • Invited
  • [Presentation] Influence of Intra-Cellular Nanoendoscopy-AFM Measurements on Cell Viability and Functions2021

    • Author(s)
      M. M. Hosain, M. S. Alam, T. Ichikawa, M. Penedo, T. Shirokawa, S Kundan, K. Miyazawa,K. Miyata, T. Fukuma
    • Organizer
      59th Annual Meeting of the Biophysical Society of Japan
    • Related Report
      2021 Annual Research Report
  • [Presentation] Development of the nanoendoscopy AFM technique for visualizing inter-cellular structures in living cells2021

    • Author(s)
      M. S. Alam, M. Penedo, T. Shirakawa, H. M. Mubarak, T. Ichikawa, K. Miyazawa, K. Miyata, C. Nakamura, T. Fukuma
    • Organizer
      59th Annual Meeting of the Biophysical Society of Japan
    • Related Report
      2021 Annual Research Report
  • [Presentation] 生細胞内部の三次元立体構造を可視化するナノ内視鏡の開発2021

    • Author(s)
      城川 哲也, Mohammad Shahidul Alam, Mohammad Mubarak Hosain, Penedo Garcia Marcos, 宮澤 佳甫, 中村 史, 福間 剛士
    • Organizer
      第82回応用物理学会秋季学術講演会
    • Related Report
      2021 Annual Research Report
  • [Presentation] 特徴的な3次元構造を抽出するための3次元AFMデータ解析手法の開発2021

    • Author(s)
      前坂 智弘, 宮下 尚之, 宮澤 佳甫, Garcia Marcos Penedo, 宮田 一輝, 福間 剛士
    • Organizer
      第82回応用物理学会秋季学術講演会
    • Related Report
      2021 Annual Research Report
  • [Presentation] 液中AFMによる界面現象や生命現象のオペランド計測と「次世代」の方向性2020

    • Author(s)
      福間剛士
    • Organizer
      JST-CRSD科学技術未来戦略ワークショップ
    • Related Report
      2020 Research-status Report
    • Invited
  • [Presentation] Visualizing Inside of 3D Self-Organizing Systems by 3D-AFM2020

    • Author(s)
      Takeshi Fukuma
    • Organizer
      The Virtual Symposium on Scanning Probe Microscopy: Current Status and Future Trends
    • Related Report
      2020 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] Visualizing Inside of 3D Self-Organizing Systems by 3D-AFM2020

    • Author(s)
      Takeshi Fukuma
    • Organizer
      Virtual RMS AFM & SPM Meeting
    • Related Report
      2020 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] Visualizing Inside of 3D Self-Organizing Systems by 3D-AFM2020

    • Author(s)
      Takeshi Fukuma
    • Organizer
      The 8th Multifrequency Conference
    • Related Report
      2020 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] Visualizing Intracellular Structures with Sub-nanometer Scale Resolution by 3D-AFM in Liquid Environment2020

    • Author(s)
      K. Miyata, M. P. Garcia, K. Miyazawa, T. Shirokawa, A. M. Shahidul, C. Nakamura, T. Fukuma
    • Organizer
      The 28th International Colloquium on Scanning Probe Microscopy (ICSPM28)
    • Related Report
      2020 Research-status Report
    • Int'l Joint Research
  • [Presentation] 細胞の三次元内部構造を可視化するナノ内視鏡の創成2020

    • Author(s)
      宮澤佳甫、Marcos Penedo Garcia、城川哲也、須崎萌、中村史、福間剛士
    • Organizer
      第81回応用物理学会秋季学術講演会
    • Related Report
      2020 Research-status Report
  • [Presentation] 液中原子間力顕微鏡技術の開発とそのナノバイオサイエンスへの応用2019

    • Author(s)
      福間剛士
    • Organizer
      第32回バイオエンジニアリング講演会
    • Related Report
      2019 Research-status Report
    • Invited
  • [Presentation] 革新的バイオSPM技術による生命科学における未踏ナノ領域の開拓2019

    • Author(s)
      福間剛士
    • Organizer
      LINK-Jネットワーキング・ナイト#40
    • Related Report
      2019 Research-status Report
    • Invited
  • [Presentation] 液中高分解能AFM技術の高速化と多機能化2019

    • Author(s)
      福間剛士
    • Organizer
      日本学術振興会ナノプローブテクノロジー第167委員会 第93回研究会
    • Related Report
      2019 Research-status Report
    • Invited
  • [Presentation] 液中原子間力顕微鏡による生きた細胞の表面および内部のナノ構造計測2019

    • Author(s)
      福間剛士
    • Organizer
      ナノプローブ表面特性評価技術セミナー
    • Related Report
      2019 Research-status Report
    • Invited
  • [Presentation] 液中表面をAFMでみる2019

    • Author(s)
      福間剛士
    • Organizer
      CSJ化学フェスタ
    • Related Report
      2019 Research-status Report
    • Invited
  • [Presentation] 2D/3D-AFM imaging of biological systems and demands for advanced AFM data analysis2019

    • Author(s)
      Takeshi Fukuma
    • Organizer
      The 57th Annual Meeting of the Biophysical Society Japan
    • Related Report
      2019 Research-status Report
    • Invited
  • [Presentation] 液中原子間力顕微鏡による2次元および3次元界面計測技術2019

    • Author(s)
      福間剛士
    • Organizer
      JASISコンファレンス「走査型プローブ顕微鏡を利用した先端分析技術」
    • Related Report
      2019 Research-status Report
    • Invited
  • [Presentation] 高分解能液中原子間力顕微鏡技術の開発とその化学分野における応用2019

    • Author(s)
      福間剛士
    • Organizer
      化学とマイクロ・ナノシステム学会
    • Related Report
      2019 Research-status Report
    • Invited
  • [Remarks] 福間研究室ホームページ

    • URL

      http://fukuma.w3.kanazawa-u.ac.jp/

    • Related Report
      2021 Annual Research Report
  • [Remarks] 金沢大学ナノ生命科学研究所ホームページ

    • URL

      https://nanolsi.kanazawa-u.ac.jp/

    • Related Report
      2021 Annual Research Report

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Published: 2019-07-04   Modified: 2023-01-30  

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