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Development of the rapid cell imaging system which can identify the chemical species without labeling by inverse Raman spectroscopy

Research Project

Project/Area Number 18K06163
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 43040:Biophysics-related
Research InstitutionNara Institute of Science and Technology

Principal Investigator

Nakashima Satoru  奈良先端科学技術大学院大学, 研究推進機構, URA(チーフ) (80263234)

Co-Investigator(Kenkyū-buntansha) 廣田 俊  奈良先端科学技術大学院大学, 先端科学技術研究科, 教授 (90283457)
Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2019: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2018: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
Keywordsラマン顕微鏡 / 逆ラマン現象 / ポンププローブ法 / 2次元検出器 / リアルタイム / 細胞ダイナミクス / 小分子 / 非染色・非侵襲 / ポンプ-プローブ法 / 2次元検出器 / 脂質モデル / 逆ラマン分光法 / 顕微
Outline of Final Research Achievements

As basic data, I constructed an optical system with a single detection element, and then acquired data on the detection limit level of this system. A microscopic optical system was constructed, and two colors of light were incident coaxially, and detections were performed in an arrangement that allows observation of a region of several tens of micro meter. Through the precise design of the optical system, I was able to develop a single-channel inverse Raman microscope that can detect intensity changes of 0.06% or less in near real time (2 seconds). Using this device, I succeeded in microscopic observation of a sample using oil droplets in an aqueous solution as a model substance for lipids by this reverse Raman measurement system. In the next step, I replaced a single element with a CMOS-type ultra-high-speed two-dimensional detector, and replaced the optical system with the aim of creating an imaging-type inverse Raman microscope.

Academic Significance and Societal Importance of the Research Achievements

細胞内のタンパク質や小分子の分布、特にその動的過程は細胞の機能を調べる上で非常に重要である。生きた細胞を非侵襲、非標識で直接観測することができれば極めて有効な手段となる。ラマン分光法は原理的にはこのような目的には最適であるが、感度が低くまた積算時間がかかることが壁となってきていた。近年格段に進歩してきた、高繰り返しレーザと高速検出器を活用して、逆ラマン分光法という非線形現象を利用することで、これらの難点を克服できると考え研究を進めた。細胞内の生命現象をライブで捉えて観察できることは、細胞生物学のステージを上げることに繋がり、がん細胞の形成過程など疾病治療への道も開ける。

Report

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

    (5 results)

All 2019 2018

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

  • [Journal Article] New Assay Method Based on Raman Spectroscopy for Enzymes Reacting with Gaseous Substrates2019

    • Author(s)
      Y. Kawahara‐Nakagawa, K. Nishikawa, S. Nakashima, S. Inoue, T. Ohta, T. Ogura, Y. Shigeta, K. Fukutani, T. Yagi, and Y. Higuchi
    • Journal Title

      Protein Science

      Volume: 28 Issue: 3 Pages: 663-670

    • DOI

      10.1002/pro.3569

    • Related Report
      2019 Research-status Report 2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Performance of a time-resolved IR facility for assessment of protonation T states and polarity changes in carboxyl groups in a large membrane protein, mammalian cytochrome c oxidase, under turnover conditions in a sub-millisecond time resolution2018

    • Author(s)
      Chen Li, Tatsuhito Nishiguchi, Kyoko Shinzawa-Itoh, Shinya Yoshikawa, Takashi Ogura, Satoru Nakashima
    • Journal Title

      Biochimica et Biophysica Acta

      Volume: 1859 Issue: 10 Pages: 1045-1050

    • DOI

      10.1016/j.bbabio.2018.05.015

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Presentation] 時間分解赤外分光法解析によるシトクロム酸化酵素の反応中間体プロトン移動機構2019

    • Author(s)
      中島聡、Li Chen、伊藤(新澤)恭子、小倉尚志、吉川信也
    • Organizer
      日本生体エネルギー研究会討論会
    • Related Report
      2019 Research-status Report
  • [Presentation] Proton Pumping Dynamics during Catalytic Cycle in Cytochrome c Oxidase Revealed by Time-Resolved IR Spectroscopy2018

    • Author(s)
      Satoru Nakashima, Chen Li, Tatsuhito Nishiguchi, Kyoko Shinzawa-Itoh, Shinya Yoshikawa, Takashi Ogura
    • Organizer
      10th International Conference on Porphyrins and Phthalocyanines
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] Proton pumping mechanism of cytochrome c oxidase by time-resolved vibrational spectroscopy2018

    • Author(s)
      Satoru Nakashima
    • Organizer
      第56回日本生物物理学会年会
    • Related Report
      2018 Research-status Report
    • Invited

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Published: 2018-04-23   Modified: 2022-01-27  

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