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Development of Super Sensitive Position Detector and Its Application on the Studies of Plasma Membrane Proteins

Research Project

Project/Area Number 08558075
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Biophysics
Research InstitutionOsaka University (1997)
The University of Tokyo (1996)

Principal Investigator

SAKO Yasushi  Medical School, Assistant Prof., 医学部, 助手 (20215700)

Co-Investigator(Kenkyū-buntansha) HORIGUCHI Chiyoharu  Hamamatsu Photonics KK.Chief, 主任研究員
Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥8,900,000 (Direct Cost: ¥8,900,000)
Fiscal Year 1997: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1996: ¥6,900,000 (Direct Cost: ¥6,900,000)
Keywordsmembrane proteins / membrane skeleton / cytoskeleton / Brownian diffusion / quadramut photodiode / 光学顕微鏡 / 変位計測 / 振動計測 / 細胞膜 / 膜タンパク質
Research Abstract

A new-type super-sensitive position detector was developed using a quadrant avalanche photodiode (4APD) as the photo sensor. This position detector was interfaced to a newly-developed reflective polarisation optical microscope with a near-JR (810 nm) external cavity diode Laser as the light source, In this system, temperature and voltage of 4APD can be optimised to achieve maximum S/N ratio.
Using colloidal gold particles with a diameter of 10-100 nm as standard samples, 0.5-2 V output signal with>100 S/N was obtained. At this range of signal intensity, bandwidth of position measurement was 0-17 MHz (-3dB).
Spatial resolution of the system was examined by moving the standard sample by a PZT stage. At 375x magnification of the microscope, spatial resolution better than 3 nm was achieved. At this time, the spatial resolution is determined by a fluctuation of the power supply, and using more stable power supply, it should be improved to more than 1 nm. Movement within a 200 nm-phi area can be traced.
Using this system, lateral movement of plasma membrane proteins were measured. Although membrane proteins which is bound to the cytoskeleton was undetectable in 33 ms video-rate time window, rapid and Browninan diffusion movement restricted within an area of 300-500 nm was observed by more than 1 ms time resolution of the newly developed position detector. Such restricted movement of cytoskeleton-bound proteins suggests flexibility of the cytoskeletal filaments.

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • Research Products

    (19 results)

All Other

All Publications (19 results)

  • [Publications] Kusumi,A.& Sako,Y.: "Cell surface organization by the membrane skeleton." Curr.Opin.Cell Biol.8. 566-574 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Sako,Y.et al: "Comparison of two-photon excitation laser scanning microscopy with UV-confocal laser scanning microscopy in 3-dimensional calcium imaging using the fluorescence indicator Indo-1" J.Microscopy. 18.5. 9-20 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Sako,Y.et al.: "Cytoplasmic regulation of the movement of E-cadherin on the free cell surface as studied by optical tweezers and single particle tracking : corralling and tethering by the membrane skeleton." J.Cell Biol.140. 1227-1240 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Takeuch,M.,Sako,Y.,et al.: "Structure of the erythrocyte membrane skeleton as observed by atmic force microscopy." Biophys.J.74. 2171-2183 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Tomishige,M.,Sako,Y.& Kusumi,A.: "Hop diffusion of band3 across the membrane skeleton barriers in the erythrocyte membrane." J.Cell Biol.142. 989-1000 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kusumi,A.,Sako,Y.,et al.: "Application of laser tweezers to studies of the fences and tethers of the membrane skeleton which regulate the movements of plasma membrane proteins." Methods Cell Biol.55. 174-194 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kusumi, A.and Sako, Y.: "Cell surface organization by the membrane skeleton." Curr.Opin.Cell Biol.8. 566-574 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Sako, Y., Sekihata, A., Yanagisawa, Y., Yamamoto, M., Shimada, Y., Ozaki, K., and Kusumi, A.: "Comparison of two-photon excitation laser scanning microscopy with UV-confocal laser scanning microscopy in 3-dimensional calcium imaging using the fluorescence indicator Indo-1." J.Microscop. 185. 9-20 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Sako, Y., Nagafuchi, A., Tsukita, S., Takeichi, M., and Kusumi, A.: "Cytoplasmic regulation of the movement of E-cadherin on the free cell surface as studied by optical tweezers and single particle tracking : corralling and tethering by the membrane skeleton." J.Cell Biol.140. 1227-1240 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Takeuchi, M., Miyamoto, H., Sako.Y., Komizu, H., and Kusumi, A.: "Structure of the erythrocyte membrane skeleton as observed by atmic force microscopy." Biophys.J.74. 2171-2183 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Tomishige, M., Sako, Y., and Kusumi, A.: "Hop diffusion of band3 across the membrane skeleton barriers in the erythrocyte membrane." J.Cell Biol.142. 989-1000 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kusumi, A., Sako, Y., Fujiwara, T., and Tomishige, M.: Application of laser tweezers to studies of the fences and tethers of the membrane skeleton which regulate the movements of plasma membrane proteins.Method.Cell Biol.55, "Laser Tweezers in Cell Biology", Sheetz, M.P.ed.Academic Press, 174-194

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Sako.Y.: "Comparison of two-photon excitation laser scanning microscopy with UV-confocal laser scanning microscopy in 3-dimensional calcium imaging using the fluorescence indicator Indo-1." J.Micros.185. 9-20 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kusumi,A.: "Application of laser tweezers to studies of the fences and tethers of the membrane skeleton which regulate the movements of plasma membrane proteins." Method.Cell Biol.55. 174-194 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Sako,Y.: "Cytoplasmic regulation of the movement of E-cadherin on the free cell surface as studied by optical tweezers and single particle tracking : corralling and tethering by the membrane skeleton." J.Cell Biol.(in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] Takeuchi,M.: "Structure of erythrocyte membrane skeleton as observed by atmic force microscopy." Biophys.J.(in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] Sako,Y.et al.: "Comparison of two-photon excitation laser scanning microscopy with UV-confocal laser scanning microscopy in 3-dimensional calcium imaging using the fluorescence indicator Indo-1" J.Microscopy. 185(in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 佐甲靖志、富重道雄、藤原敬宏: "光ピンセット法とその細胞膜研究への応用" ファルマシア. 32. 271-275 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 佐甲靖志: "光学顕微鏡で見る分子の世界" 「電子顕微鏡 基礎技術と応用1996」. 135-142 (1996)

    • Related Report
      1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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