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Time-resolved spectroscopy of a specific, single fluorescent protein molecule in a living cell

Research Project

Project/Area Number 17360031
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied optics/Quantum optical engineering
Research InstitutionOsaka University

Principal Investigator

ITOH Kazuyoshi  Osaka University, Graduate School of Engineering, Professor (80113520)

Co-Investigator(Kenkyū-buntansha) KONISHI Tsuyoshi  Osaka University, Graduate School of Engineering, Assoc. Prof. (90283720)
MATSUNAGA Sachihiro  Osaka University, Graduate School of Engineering, Lecturer (40323448)
OZEKI Yasuyuki  Osaka University, Graduate School of Engineering, Assist. Prof. (60437374)
渡辺 歴  独立行政法人産業技術総合研究所, 光技術研究部門, 研究員 (90314377)
Project Period (FY) 2005 – 2006
Project Status Completed (Fiscal Year 2006)
Budget Amount *help
¥15,000,000 (Direct Cost: ¥15,000,000)
Fiscal Year 2006: ¥9,500,000 (Direct Cost: ¥9,500,000)
Fiscal Year 2005: ¥5,500,000 (Direct Cost: ¥5,500,000)
KeywordsLiving cells / femtosecond laser / fluorescent protein / nonlinear optics / stimulated parametric emission / 顕微鏡 / 生体分子追跡 / タンパク質 / 干渉 / 4光波混合 / 超短光パルス / 3次元走査 / 分光画像 / 生細胞観察 / 可視化
Research Abstract

In the recent post-genome biological research field, there is a growing demand for exploring the function, mechanism, interaction, and network of proteins, which are the physical basis of life. In particular, it is of great interest to analyze the interactions among protein molecules by use of low-invasive visualization tools.
The objective of this research was to provide optical means for analyzing the interactions among protein molecules. The first target was to improve the fluorescence yield of fluorescent proteins through the investigation on the fluorescence mechanisms of fluorescent proteins with the femtosecond time-resolved micro-spectroscopy. Through the experiment, we succeeded in the two-photon fluorescence conversion of a fluorescent protein Kaede. This technique turned out to be a powerful tool for exploring the function of a single organelle in a living cell. Then we realized that, by irradiating dual-wavelength femtosecond pulses, a strong optical signal is generated through the four-wave mixing process due to electronic optical nonlinearity, which we called stimulated parametric emission, SPE. We constructed a SPE microscopy system, which was found to be a useful tool for the label-free observation of living cells. We also succeeded in (a) the sensitive detection of SPE signals by use of a spectral interferometric technique, (b) the application of the SPE microscopy to the three-dimensional profiling of refractive index distribution of non-biomedical samples, and (c) the development of a novel technique of dispersion compensation of a microscope objective lens for its application to ultrafast nonlinear optical microscopy using ultrashort optical pulses in few-cycle regime. To summarize, through these achievements, we have successfully developed and demonstrated novel approaches for exploring interactions of protein molecules by using femtosecond laser pulses.

Report

(3 results)
  • 2006 Annual Research Report   Final Research Report Summary
  • 2005 Annual Research Report
  • Research Products

    (13 results)

All 2007 2006

All Journal Article (11 results) Patent(Industrial Property Rights) (2 results)

  • [Journal Article] Single-organelle tracking by two-photon conversion2007

    • Author(s)
      W. Watanabe, et al.
    • Journal Title

      Optics Express 15

      Pages: 2490-2498

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Single-organelle tracking by two-photon conversion2007

    • Author(s)
      W. Watanabe, T. Shimada, S. Matsunaga, D. Kurihara, K. Fukui, S. -i. Arimura, N Tsutsumi, K. Isobe, K. Itoh
    • Journal Title

      Opt. Express vol. 15

      Pages: 2490-2498

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Single-organelle tracking by two-photon conversion.2007

    • Author(s)
      Watanabe, W., Shimada, T., Matsunaga, S., Kurihara, D., Fukui, K., Arimura, S., Tsutsumi, N., Isobe, K. and Itoh, K.
    • Journal Title

      Optics Express 15

      Pages: 2490-2490

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Stimulated parametric emission microscopy2006

    • Author(s)
      K. Isobe, et al.
    • Journal Title

      Optics Express 14

      Pages: 786-793

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Highly sensitive spectral interferometric four-wave mixing microscopy near the shot noise limit and its combination with two-photon excited fluorescence microscopy2006

    • Author(s)
      K. Isobe, et al.
    • Journal Title

      Optics Express 14

      Pages: 11204-11214

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Stimulated parametric emission microscopy2006

    • Author(s)
      K. Isobe, S. Kataoka, R. Murase, W. Watanabe, T. Higashi, S. Kawakami, S. Matsunaga, K. Fukui, K. Itoh
    • Journal Title

      Opt. Express vol. 14

      Pages: 786-793

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Highly sensitive spectral interferometric four-wave mixing microscopy near the shot noise limit and its combination with two-photon excited fluorescence microscopy2006

    • Author(s)
      K. Isobe, Y. Ozeki, T. Kawasumi, S. Kajiyama, K. Fukui, K. Itoh
    • Journal Title

      Opt. Express vol. 14, no. 23

      Pages: 11204-11214

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Stimulated parametric emission microscopy2006

    • Author(s)
      K. Isobe, S. Kataoka, R. Murase, W. Watanabe, T. Higashi, S. Kawakami, S. Matsunaga, K. Fukui, and K. Itoh
    • Journal Title

      Optics Express 14

      Pages: 786-786

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Highly sensitive spectral interferometric four-wave mixing microscopy near the shot noise limit and its combination with two-photon excited fluorescence microscopy2006

    • Author(s)
      K. Isobe, Y. Ozeki, T. Kawasumi, S. Kataoka, S.'i. Kajiyama, K. Fukui, and K. Itoh
    • Journal Title

      Optics Express 14

      Pages: 11204-11204

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Coherence-based 3-D and spectral imaging and laser-scanning microscopy2006

    • Author(s)
      K. Itoh, W. Watanabe, H. Arimoto, K. Isobe
    • Journal Title

      Proceedings of IEEE 94

      Pages: 608-608

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Stimulated parametric emission microscopy2006

    • Author(s)
      Keisuke Isobe
    • Journal Title

      Optics Express 14・2

      Pages: 786-793

    • Related Report
      2005 Annual Research Report
  • [Patent(Industrial Property Rights)] 空間情報検出装置2006

    • Inventor(s)
      伊東一良ほか4名
    • Industrial Property Rights Holder
      大阪大学
    • Industrial Property Number
      2006-307004
    • Filing Date
      2006
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Patent(Industrial Property Rights)] 屈折率測定方法および屈折率測定装置, 光学部材の製造方法2006

    • Inventor(s)
      伊東一良ほか5名
    • Industrial Property Rights Holder
      大阪大学
    • Industrial Property Number
      2006-302038
    • Filing Date
      2006
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary

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Published: 2005-04-01   Modified: 2016-04-21  

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