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Single-molecule fluorescence detection of kinesin and ATP using nanotracks

Research Project

Project/Area Number 16K13644
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Nanobioscience
Research InstitutionKyoto University

Principal Investigator

Yokokawa Ryuji  京都大学, 工学研究科, 准教授 (10411216)

Co-Investigator(Renkei-kenkyūsha) IINO Ryota  分子科学研究所, 生命・錯体分子科学研究領域 生体分子機能研究部門, 教授 (70403003)
TOMISHIGE Michio  青山学院大学, 理工学部, 教授 (50361530)
Project Period (FY) 2016-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Keywordsマイクロ・ナノデバイス / 分子モーター / 生物物理 / ナノバイオ / 1分子計測(SMD) / 1分子計測(SMD)
Outline of Final Research Achievements

We developed Linear Zero Mode Waveguides (LZMWs) for simultaneous observation of kinesin and ATP under higher concentration of fluorescently labeled ATP than that used in the conventional total internal reflection microscopy (TIRFM). Single-molecule fluorescent microscopy is a promising method to analyze correlation between the mechanical displacement and hydrolysis of ATP by a kinesin molecule. However, the concentration of fluorescently labeled ATP was limited up to 10 nM in TIRFM, thus available data from experiment was limited. Proposed method enables to use ten times higher concentration of fluorescently labeled ATP by confining excitation lights in LZMWs, which are nano-slits with 100 nm in width.

Report

(3 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • Research Products

    (17 results)

All 2018 2017 2016 Other

All Journal Article (1 results) (of which Peer Reviewed: 1 results) Presentation (15 results) (of which Int'l Joint Research: 7 results,  Invited: 5 results) Remarks (1 results)

  • [Journal Article] Design and Fabrication of Linear-shaped Zero Mode Waveguides for Single Molecule Observation of Kinesin and Fluorescent ATP2017

    • Author(s)
      Fujimoto Kazuya、Morita Yuki、Iino Ryota、Tomishige Michio、Shintaku Hirofumi、Kotera Hidetoshi、Yokokawa Ryuji
    • Journal Title

      IEEJ Transactions on Sensors and Micromachines

      Volume: 137 Issue: 6 Pages: 159-164

    • DOI

      10.1541/ieejsmas.137.159

    • NAID

      130005687854

    • ISSN
      1341-8939, 1347-5525
    • Related Report
      2017 Annual Research Report
    • Peer Reviewed
  • [Presentation] Fluorescent observation of ATP binding in kinesin driven microtubule gliding using nano-slits2018

    • Author(s)
      K. Fujimoto, R. Iino, M. Tomishige, H. Shintaku, H. Kotera, R. Yokokawa
    • Organizer
      The 31st IEEE International Conference on Micro Electro Mechanical Systems (MEMS 2018)
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Micro/nano fabrications for biophysics and organ-on-a-chip applications2018

    • Author(s)
      R. Yokokawa
    • Organizer
      IEEE Electron Devices Technology and Manufacturing (EDTM) Conference
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Linear Zero Mode Waveguides for the Study of Chemo-Mechanical Coupling Mechanism of Kinesin2017

    • Author(s)
      K. Fujimoto, Y. Morita, R. Iino, M. Tomishige, H. Shintaku, H. Kotera, and R. Yokokawa
    • Organizer
      The 30th IEEE International Conference on Micro Electro Mechanical Systems (MEMS 2017)
    • Place of Presentation
      Las Vegas, USA
    • Year and Date
      2017-01-23
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] キネシンによる微小管運動における蛍光ATP結合・解離のLZMWsを用いた1分子観察2017

    • Author(s)
      藤本 和也, 飯野 亮太, 富重 道雄, 新宅 博文, 小寺 秀俊, 横川 隆司
    • Organizer
      第34回「センサ・マイクロマシンと応用システム」シンポジウム
    • Related Report
      2017 Annual Research Report
  • [Presentation] ナノスリットを用いたキネシン・微小管および蛍光ATPの同時1分子観察2017

    • Author(s)
      藤本 和也, 飯野 亮太, 富重 道雄, 新宅 博文, 小寺 秀俊, 横川 隆司
    • Organizer
      第36回化学とマイクロ・ナノシステム学会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Micro/nano fabrications for biophysical studies of endothelial cells and motor proteins2017

    • Author(s)
      R. Yokokawa
    • Organizer
      International Conference on Molecular Electronics and Bioelectronics (M&BE9)
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Micro/nano fabrications for engineering applications and biophysical studies of motor proteins2016

    • Author(s)
      R. Yokokawa
    • Organizer
      International Conference on BioSensors, BioElectronics, BioMedical Devices, BioMEMS/NEMS & Applications (Bio4Apps 2016)
    • Place of Presentation
      Gold Coast, Australia
    • Year and Date
      2016-12-14
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] LZMWを利用した高濃度蛍光ATP存在下でのキネシン運動とATP結合の同時蛍光1分子観察2016

    • Author(s)
      藤本 和也, 森田 有貴, 新宅 博文, 飯野 亮太, 富重 道雄, 小寺 秀俊, 横川 隆司
    • Organizer
      第54回日本生物物理学会
    • Place of Presentation
      つくば
    • Year and Date
      2016-11-25
    • Related Report
      2016 Research-status Report
  • [Presentation] Single-Molecule Fluorescence Imaging of Kinesin Using Linear Zero-Mode Waveguides2016

    • Author(s)
      Y. Morita, K. Fujimoto, R. Iino, M. Tomishige, H. Shintaku, H. Kotera, R. Yokokawa
    • Organizer
      IEEE Sensors 2017
    • Place of Presentation
      Orlando, USA
    • Year and Date
      2016-10-30
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] LZMWを用いたキネシンによるATP加水分解サイクルの可視化2016

    • Author(s)
      藤本 和也, 森田 有貴, 新宅 博文, 飯野 亮太, 富重 道雄, 小寺 秀俊, 横川 隆司
    • Organizer
      第33回「センサ・マイクロマシンと応用システム」シンポジウム
    • Place of Presentation
      平戸
    • Year and Date
      2016-10-24
    • Related Report
      2016 Research-status Report
  • [Presentation] Linear Zero Mode WaveguideによるキネシンのATP加水分解サイクル1分子計測2016

    • Author(s)
      藤本 和也, 森田 有貴, 新宅 博文, 飯野 亮太, 富重 道雄, 小寺 秀俊, 横川 隆司
    • Organizer
      第67回コロイドと界面化学討論会
    • Place of Presentation
      旭川
    • Year and Date
      2016-09-22
    • Related Report
      2016 Research-status Report
  • [Presentation] R14Aキネシンによるナノトラックへの微小管の固定法とその評価2016

    • Author(s)
      森田 有貴, 藤本 和也, 飯野 亮太, 富重 道雄, 新宅 博文, 小寺 秀俊, 横川 隆司
    • Organizer
      化学とマイクロ・ナノシステム学会 第34回研究会
    • Place of Presentation
      千葉
    • Year and Date
      2016-09-06
    • Related Report
      2016 Research-status Report
  • [Presentation] キネシン1分子観察系構築に向けたLZMWへの微小管の固定2016

    • Author(s)
      森田 有貴, 藤本 和也, 飯野 亮太, 富重 道雄, 新宅 博文, 小寺 秀俊, 横川 隆司
    • Organizer
      平成28年度電気学会 センサ・マイクロマシン部門総合研究会 バイオ・マイクロシステム研究会
    • Place of Presentation
      金沢
    • Year and Date
      2016-06-29
    • Related Report
      2016 Research-status Report
  • [Presentation] 微小流体デバイスのタンパク質・細胞応用2016

    • Author(s)
      横川隆司
    • Organizer
      精密工学会「第4回IoT製造研究会」
    • Place of Presentation
      京都
    • Year and Date
      2016-06-24
    • Related Report
      2016 Research-status Report
    • Invited
  • [Presentation] Microfluidic devices for biophysical studies and engineering applications of motor proteins2016

    • Author(s)
      R. Yokokawa
    • Organizer
      OIST Mini Symposium; Science and Technology at the Interface of Bio-Nano-Systems: Challenges and Opportunities
    • Place of Presentation
      Onna, Japan
    • Year and Date
      2016-04-25
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research / Invited
  • [Remarks] 横川 隆司の個人ページ

    • URL

      http://www.ksys.me.kyoto-u.ac.jp/ry/

    • Related Report
      2016 Research-status Report

URL: 

Published: 2016-04-21   Modified: 2019-12-27  

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