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エバネッセント光を利用した分子間力3次元測定可能な光学顕微鏡の試作

Research Project

Project/Area Number 07558100
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section試験
Research Field Biophysics
Research InstitutionTohoku university (1996)
Keio University (1995)

Principal Investigator

MIYATA Hidetake  Tohoku university Graduate School of Science, Physics Department, Associate professor, 大学院・理学研究科, 助教授 (90229865)

Co-Investigator(Kenkyū-buntansha) ITOH Hiroyasu  Hamamatsu Photonics, .Inc., Tukuba.Research Institute, Investigator, 筑波研究所, 研究員
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1996: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1995: ¥5,500,000 (Direct Cost: ¥5,500,000)
Keywordsoptical tweezers / evanescent wave / optical microscope / intermolecular forces / エバネッセント / イメージング
Research Abstract

The major goal of our project was to establish a methodology which enables us to simultaneously image and manipulate single molecules under an optical microscope. We attempted to combine two techniques, namely, an optical trapping and evanescent excitation microscopy. To do this, we drastically modified an old methodology to create the evanescent field at glass-water interface : we utilized a coverslip as a waveguide so that a YAG or an argon ion laser beam excuse total internal reflection multiple times and reach the desired position of the coverslip where sample is located. This methodology was anticipated to be coupled to the optical trapping technique in less interfering manner than the conventional method to create the evanescent field. The optical trapping and the new evanescent excitation method satisfactorily well : we could measure the actomyosin motor force, cohesive force between actin and alpha-actinin, or we could observe single fluorescent molecules at the glass-water interface. When the two methods combined, a problem which was not anticipated before appeared : aberration of the objective lens against the infra-red laser beam used for the optical trap was far stronger than our anticipation and it was almost impossible to use optical trap under the setup we utilized. We hence searched other ways, but found the problem should be resolved by designing a new objective lens with aberration is corrected for infra-red wavelength region. At present, the usage of the infra-red wavelength light as a excitation light source for fluorescence microscopy, close cooperation with microscope manufacturer will be the most urgent to approach and achieve our goal.

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (23 results)

All Other

All Publications (23 results)

  • [Publications] Hidetake Miyata etal.: "Evanescent excitation microscopy:its application to the study of single molecular process kinetics of actomyosin motor" The proceedings of the Symposium on Mechanism of Work Production and Work Absroption in Muscle. in press. (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 宮田 英威: "光ピンセット法" 蛋白質 核酸 酵素増刊号「見る技術」. 42. 1170-1172 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hiroyosu Itoh etal.: "Flurorescence lifetime imaging microscopy with a high repetition gated camera and a dual-view assembly for the real time measurement" Proceedings of SPIE. 2980. 12-19 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 佐瀬 一郎 他: "蛍光色素1分子イメージング" ファルマシア. 32. 281-284 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hidetake Miyata etal.: "Strength and bond lifetime of actin-skeletal muscle α-actinin bonds studied by an optical trapping technique." Biochimica et Biophysica Acta. 1290. 83-88 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hidetake Miyata etal: "Mechanical measurements of single actomyosin motor force" Biophysical Journal. 68. 286S-290S (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 木下 一彦 他: "「バイオイメージング」中 第1章5節 光ピンセット-1分子を見て操作する" 共立出版社, 266 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Miyata, H., Chaen, S., Sugiura, S.and Sugi, H.: " "Evanescent excitation microscopy : its application to the study of single molecular process kinetics of actomyosin motor"" Proceedings of the Symposium on Mechanism of Work Production and Work Absorption in Muscle. (in press). (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Miyata, H., Yasuda, R.and Kinoshita, K., Jr.: " "Strength and bond lifetime of action-skeletal muscle alpha-actinin bonds studied by an optical trapping technique"" Biochimica Biophysica Acta. 1290. 83-88 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Itho, H., Evenzahav, A., Inagaki, Y., Mizushima, H.Takahashi, A., Fukami, T., Hayakawa, T., and Kusumi, A.: " "Fluorescence lifetime imaging microscopy with a high repetition gated camera and a dual-view assembly for the real time measurement"" Proceedings of SPIE. 2980. 12-19 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Miyata, H., Yoshikawa, H., Hakozaki, H., SUZUKI,N., FURUNO,T., IKEGAMI,A., Kinosita, K.Jr., Nishizaka, T.and Ishiwata, S.: " "Mechanical measurements of single actomyosin motor force"" Biophysical Journal. 68. 286S-290S (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 宮田英威: "光ピンセット法" 蛋白質 核酸 酵素 増刊号「見る技術」. (掲載予定). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hidetake Miyata 他: "Cooperative associatoin of actin protomers and actin oligomers at low ionic strength" Journal of Biochemistry. 1・3(掲載予定). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 佐瀬一郎 他: "個を見る" 日本生物物理学会誌. 36・1. 20-24 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Ken-ichirou Akashi 他: "Preparation of giant liposomes in physiological conditions and their characterization under an optical microscope" Biophysical Journal. 71・6. 3242-3250 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hidetake Miyata 他: "Strength and bond lifetime of actin-skeletal muscle αactinin bonds studied by an optical trapping technique" Biochimica et Biophysica Acta. 1290・1. 83-88 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Ryohei Yasuda 他: "Direct measurement of the tortional rigidity of single actin filaments" Journal of Molecular Biology. 263・2. 227-236 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Miyata,H.,et al.,: "Mechanical measurements of single actomyosin motor force." Biophys.J.68. 286s-290s (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Sase,I.,et.al.,: "Realtime imaging of single fluorophores on moving actin with an ordinary epifluorescence microscope." Biophys.J.69. 323-328 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Sase,I.,et al.,: "Regulatory mechanism of acrosome reaction revealed by multiview microscopy of single starfish sperm." J.Cell Biol.131. 963-973 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Suzuki,N.,et al.: "Preparation of bead-tailed F-actin: Estimation of the torque component of the sliding force in an in vitro motility assay." Biophys.J.70. 401-408 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Nishizaka,T.,et al.: "Unbinding force of a single motor molecule of muscle measured using optiocal tweezers." Nature. 377. 251-254 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Miyata,H.,et al.: "Strength and life time of actin-skeletal muscle α-actinin bonds studied by an optical trapping technique." B.B.A.(in press).

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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