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Three dimensional dynamics of apical membrane ruffles on living cells by scanning ion conductance microscopy

Research Project

Project/Area Number 17K15541
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field General anatomy (including histology/embryology)
Research InstitutionHokkaido University (2019-2020)
Niigata University (2017-2018)

Principal Investigator

Mizutani Yusuke  北海道大学, 総合IR室, 特任准教授 (40646238)

Project Period (FY) 2017-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2018: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Keywordsイオンコンダクタンス顕微鏡 / 3次元ダイナミクスイメージング / 細胞質突起 / 細胞骨格 / 走査型イオンコンダクタンス顕微鏡 / アクチン線維
Outline of Final Research Achievements

Scanning Ion Conductance Microscopy (SICM) is the family of scanning probe microscopy. This technique uses a glass nanopipette filled with an electrolyte as a probing tip and regulates the tip-surface distance by detecting the ion current passing through a small hole of the pipette. Thus, SICM can obtain images of three-dimensional structures in liquid conditions without mechanical contact. In this study, we succeeded in establishing a method for observing the three-dimensional dynamics of microvillous structures including dorsal ruffles on the cell membrane surface.
By using SICM and fluorescence microscopy, we observed the dynamics of the dorsal surface ruffles and the intracellular cytoskeleton related to cell motility.  Furthermore, we observed the dorsal surface ruffles of cancer cells, and showed that the structure and dynamics of dorsal surface ruffles differ greatly depending on the grade of cancer cells.

Academic Significance and Societal Importance of the Research Achievements

これまで,走査型電子顕微鏡(SEM)や生物試料の処理方法の発達に依り,高い分解能で細胞膜表面の微細構造の観察が行われてきたが,生きた細胞運動の経時的変化の観察は困難であった。本研究では,変位がドラスティックであり,膜の表面積の拡大,足場のセンシングなどに重要であると考えられている細胞質突起と呼ばれる細胞の頂上膜面に存在する微細構造を液中で生きたまま観察することを可能にした。
また,悪性度の異なるがん細胞において細胞質突起のダイナミクスが大きく異なることが明らかになったことから,細胞診断や細胞に対する薬剤の評価の新たな手法の基盤となる一助となることを示すものである。

Report

(5 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Research-status Report
  • 2018 Research-status Report
  • 2017 Research-status Report
  • Research Products

    (12 results)

All 2021 2020 2019 2018 2017 Other

All Int'l Joint Research (1 results) Journal Article (2 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 2 results,  Open Access: 1 results) Presentation (9 results) (of which Int'l Joint Research: 2 results,  Invited: 3 results)

  • [Int'l Joint Research] Park Systems Corp.(韓国)

    • Related Report
      2019 Research-status Report
  • [Journal Article] Scanning ion-conductance microscopy with a double-barreled nanopipette for topographic imaging of charged chromosomes2021

    • Author(s)
      Iwata Futoshi、Shirasawa Tatsuru、Mizutani Yusuke、Ushiki Tatsuo
    • Journal Title

      Microscopy

      Volume: - Issue: 5 Pages: 423-435

    • DOI

      10.1093/jmicro/dfab009

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] SLPI facilitates cell migration by regulating lamellipodia/ruffles and desmosomes, in which Galectin4 plays an important role2020

    • Author(s)
      Mizutani Yusuke、Omagari Daisuke、Hayatsu Manabu、Nameta Masaaki、Komiyama Kazuo、Mikami Yoshikazu、Ushiki Tatsuo
    • Journal Title

      Cell Adhesion & Migration

      Volume: 14 Issue: 1 Pages: 195-203

    • DOI

      10.1080/19336918.2020.1829264

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] Scanning ion conductance microscopy for visualizing the surface topography of rat tissue sections2021

    • Author(s)
      Mizutani Yusuke、Hayatsu Manabu、Mikami Yoshikazu、Ushiki Tatsuo
    • Organizer
      第126回日本解剖学会総会・全国学術集会
    • Related Report
      2020 Annual Research Report
  • [Presentation] イオン伝導顕微鏡による組織切片表面の微細構造イメージング2020

    • Author(s)
      水谷祐輔,早津学,三上剛和,牛木辰男
    • Organizer
      第125回日本解剖学会総会・全国集会
    • Related Report
      2019 Research-status Report
  • [Presentation] 走査型イオン伝導顕微鏡を用いた組織切片表面微細構造解析2019

    • Author(s)
      水谷祐輔,山川明里,早津学,牛木辰男
    • Organizer
      第75回日本顕微鏡学会学術講演会
    • Related Report
      2019 Research-status Report
  • [Presentation] 走査型イオン伝導顕微鏡による組織・細胞観察2019

    • Author(s)
      水谷祐輔,牛木辰男
    • Organizer
      2019年度精密工学会春季大会学術講演会
    • Related Report
      2018 Research-status Report
    • Invited
  • [Presentation] イオン伝導顕微鏡による細胞・組織イメージング2019

    • Author(s)
      水谷祐輔,早津学,三上剛和,牛木辰男
    • Organizer
      第124回日本解剖学会総会・全国集会
    • Related Report
      2018 Research-status Report
    • Invited
  • [Presentation] Simultaneous observation of the cell surface structure and cytoskeleton by scanning ion conductance microscopy and fluorescence microscopy2018

    • Author(s)
      Y. Mizutani, Y. Mikami, M. Hayatsu, T. Ushiki
    • Organizer
      The 20th International Scanning Probe Microscopy Conference (ISPM 2018)
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] 細胞表面微細構造と移動能の関連性についての走査型イオン伝導顕微鏡による解析2018

    • Author(s)
      水谷祐輔,三上剛和,牛木辰男
    • Organizer
      第74回日本顕微鏡学会学術講演会
    • Related Report
      2018 Research-status Report
  • [Presentation] 走査型イオン伝導顕微鏡による細胞膜表面観察とがん細胞動態解析への応用2018

    • Author(s)
      水谷祐輔、三上剛和、早津学、牛木辰男
    • Organizer
      第123回日本解剖学会総会・全国集会
    • Related Report
      2017 Research-status Report
    • Invited
  • [Presentation] Movement of oral carcinoma cells investigated by scanning ion conductance microscopy2017

    • Author(s)
      Y. Mizutani, Y. Yamada, Y. Mikami, T. Ushiki
    • Organizer
      The 19th International Scanning Probe Microscopy Conference (ISPM 2017)
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research

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Published: 2017-04-28   Modified: 2022-01-27  

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