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Multiple snapshots of the dynamics of supramolecular systems

Research Project

Project/Area Number 01440095
Research Category

Grant-in-Aid for General Scientific Research (A)

Allocation TypeSingle-year Grants
Research Field 生物物性学
Research InstitutionKeio University

Principal Investigator

KINOSHITA Kazuhiko  Keio University, Faculty of Science and Technology, Professor, 理工学部, 教授 (30124366)

Co-Investigator(Kenkyū-buntansha) KOUYAMA Tsutomu  Riken (The Institute for physical and chemical research), Scientist, 生物物理研究室, 研究員 (30170210)
Project Period (FY) 1989 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥33,300,000 (Direct Cost: ¥33,300,000)
Fiscal Year 1991: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1990: ¥9,400,000 (Direct Cost: ¥9,400,000)
Fiscal Year 1989: ¥21,900,000 (Direct Cost: ¥21,900,000)
KeywordsMulti-shot pulsed laser microscope / Multiview microscopy / Fluorescence polarization imaging / Cell eletroporation / Cell celctrodeformation / Actomyosin motor / Molecular mechanism of fertilization / 顕微鏡 / アクチン / ミオシン / ねじれ運動 / 受精 / ウニ / パルスレ-ザ- / 電気穿孔 / 超高速撮像技術 / リポソ-ム / 電気変形 / 燐光偏光解消 / 偏光蛍光
Research Abstract

(1) Multi-shot pulsed laser microscope
A microscope system has been developed that allows sequential capturing of four images separated by microsecond intervals. The system is based on a multi-frame streak camera and a multi-shot pulsed laser.
(2) Cell electroporation and cell electrodeformation
Under the pulsed laser above, electroporation of sea urchin eggs and of giant liposomes was investigated. The major findings were a localized increase in the membrane conductance in the order of 10 S/cm^2, and a marked asymmetry in the conductance between the anode and cathode sides of the cell. We have also found that cells and liposomes are deformed under an electric field and that its direction depends on the ratio of the intracellular and extracellular conductances.
(3) Multiview microscopy
A simple device has been developed that allows projection of differnt kinds of microscopic images (e.g., transmission and fluorescence, fluorescence at different wavelengths, polarized components of fluorescence) on a single video camera simultaneously and continuously.
(4) Measurements of molecular orientations
Using (3) above, orientations of actin monomers in and actin filament were determined. The change in orientation upon interaction with myosin was found to be small. By decreasing the number of fluorophores in a filament to one or two, we have been able to detect quantitatively the aial rotation of a sliding actin filament.
(5) Analysis of fertilization processes
Using (3), multiple fertilization processes in an egg or sperm, the changes in morphology, intracellular calcium concentration, intracellular pH,protrusion formation (egg microvilli or sperm acrosome) have been correlated spatially and temporally.

Report

(4 results)
  • 1991 Annual Research Report   Final Research Report Summary
  • 1990 Annual Research Report
  • 1989 Annual Research Report
  • Research Products

    (59 results)

All Other

All Publications (59 results)

  • [Publications] T.Koyama,et al.: "Turnover rate of the proton pumping cycle of bacteriorhodopsin ; pH and light intensity dependences." Biochemistry. 28. 5963-5970 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] H.Itoh,et al.: "Multi-shot pulsed laser fluorescence microscope." Proc.SPIE. 1204. 49-53 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] K.Uehara,et al.: "Oriented purple membrane as an ether sensor." Denki Kagaku. 58. 1132-1135 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] T.Kobayashi,et al.: "Femtosecond spectroscopy of acidified and neutral bacteriorhodopsin." Proc.SPIE. 1403. 407-416 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] K.Fukuda,et al.: "Effect of partial delipidation on the photodynamics of bacteiorhodopsin." Biochemstry. 29. 1997-2002 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] K.Uehara,et al.: "Oriented purple membrane sandwiched by proton conducting poly(ethylenimine)films and its photoresponse." Chem.Express. 5. 321-324 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] K.Imura,et al.: "Location of membrane-bound hapten with different length spacers." Immunology. 69. 323-328 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] A.Nasuda-Kouyama,et al.: "Effect of a llght-induced pH gradient on purple-to-blue and purple-to-red transitions of bacteriorhodopsin." Biochemistry. 29. 6778-6790 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] H.Hyuga,et al.: "Transient and steadey-state diformations of a vesicle with an insulating membrane in response to step-function and alternating electric fields." Jpn.J.Appl.Phys.30. 2649-2656 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 伊藤博康 他: "見えないほど速いものが...見えてくる-高時間分解能蛍光顕微鏡の開発-" バウンダリ-(コンパス社.東京). 7. 6-9 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] M.Hibino,et al.: "Membrane conductance of an electroporated cell analyzed by subicrosecond imaging of transmembrane potential." Biophys.J.59. 209-220 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] H.Hyuga,et al.: "Defomation of vesicles under the influence of strong electric fields." Jpn.J.Appl.Phys.30. 1141-1148 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] H.Hyuga,et al.: "Deformation of vesicles under the influence of strong electric fields II." Jpn.J.Appl.Phys.30. 1333-1335 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] K.Kinosita,Jr.,et al.: "Dual-view microscopy with a single camera : Real-time imaging of molecular orientations and calcium." J.Cell Biol.115. 67-73 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] K.Kimura,et al.: "Fluorescence quenching measurements of the membrane bound lipid haptens with different length spacers." Biochim.Biophys.Acta.1104. 9-14 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 神山勉: "光駆動プロトンポンプ・生体膜における物質輸送についての考察" 日本の科学と技術. 262. 61-65 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 伊藤愽康 他: "マルチショット・パルスレーザー蛍光顕微鏡による高速細胞現象のイメージング" 計測自動制御学会論文集. 28 NO.1. 156-157 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 伊藤愽康 他: "高時間分解能蛍光顕微鏡の開発マルチショット・パルスレーザー蛍光顕微鏡による細胞の高速刺激応答の観測" 比較生理生化学. 9 NO.1. 22-27 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] K.Kinosita,Jr.,et al.: "Submicrosecond imaging under a pulsed-laser fluorescence microscope. Electroporation of Cell membrane time-and space-resolved. in"Science on Form"(S.Ishizaka,ed.)" KTK Scientific Publishers(Tokyo), 11 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 木下一彦 他: "蛍光、蛍光偏光 新基礎生化学実験法5(高次構造状態分析)中嶋暉・野本明男・松橋通生・三浦謹一郎・村松正美 編" 丸善(東京), 16 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 木下一彦: "一瞬を見る(パルスレーザー顕微鏡) 限界を越える生物顕微鏡-見えないものを見る 宝谷紘一・木下一彦 編" 学会出版センター東京, 177 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] K.Kinoshita,Jr.,et al.: "Electroporation and Electrofusion. in"Events of membrane electroporation visualezed on a time scale from microsecond to seconds"(D.C.Chang et al.,eds.)" Academic Press(New York), 18 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] T.Kouyama: "Turnover rate of the proton pumping cycle of bacteriorhodopsin : pH and light intensity dependencs." Biochmistry. 28. 5963-5970 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] H.Itoh: "Multi-shot pulsed laser fluorescence microscope." Proc.SPIE. 1204. 49-53 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] K.Uehara: "Oriented purple membrane as an ether sensor." Denki Kagaku. 58. 1132-1135 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] T.Kobayashi: "Femtosecond spectroscopy of acidified and neutral bacteriorhodopsin." Proc.SPIE. 1403. 407-416 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] K.Fukuda: "Effect of partial delipidation on the photodynamics of bacteriorhodopsin." Biochemistry. 29. 1997-2002 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] K.Uehara: "Oriented purple membrane sandwiched by proton conducting poly (ethylenimine) films and its photoresponse." Chem.Express. 5. 321-324 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] K.Kimura: "Location of membrane-bound hapten with different length spacers." Immunology. 69. 323-328 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] A.Nasuda-Kouyama: "Effect of a light-induced pH gadient on purple-toblue and purple-to-red transitions of bacteriorhodopsin." Biochemistry. 29. 6779-6790 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] H.Hyuga: "Transient and steady-state deformations of a vesicle with an insulating membrane in response to step-function and alternating electric fiedlds." Jpn.J.Appl.Phys. 30. 2649-2656 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] M.Hibino: "Membrane conductance of an electroporated cell analyzed by subicrosecond imaging of transmembrane potential." Biophys.J.59. 209-220 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] H.Hyuga: "Deformation of vesicles under the influence of strong electric fields." Jpn.J.Appl.Phys.30. 1141-1148 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] H.Hyuga: "Deformation of vesicles under the influence of strong electric fields II." Jpn.J.Appl.Phys.30. 1333-1335 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] K.Kinosita, Jr.: "Dual-view microscopy with a single camera : Real-time imaging of molecular orientations and calcium." J.Cell Biol.115. 67-73 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] K.Kimura: "Fluorescence quenching measurements of the membrane bound lipid haptens with different length spacers." Biochim.Biophys.Acta. 1104. 9-14 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] K.Kinosita, Jr.: Submicrosecond imaging under a pulsed-laser fluorescence microscope.Electroporation of cell membrane time-and space-resolved.in "Science on Form" (S.Ishizaka, ed.). KTK Scientific Publichers (Tokyo), 94-104 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] K.Kinosita, Jr.: Electroporation and Electrofusion.in "events of membrane electroporation visualized on a time scale from microsecond to seconds" (D.C.Chang et al., eds.). Academic Press (New York), 29-46 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] H.Hyuga et al.: "Deformation of vesicles under the influence of strong electric fields" Jpn.J.Appl.Phys.30. 1141-1148 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] H.Hyuga et al.: "Deformation of vesicles under the influence of strong electric fields.II." Jpn.J.Appl.Phys.30. 1333-1335 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] K.Kinosita,Jr.et al.: "Dualーview microscopy with a single camera:realーtime imaging of molecular orientations and calcium" J.Cell Biol.115. 67-73 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] H.Hyuga et al.: "Transient and steadyーstate deformations of a vesicle with an insulating membrane in response to stepーfunction and alternating electric fields." Jpn.J.Appl.Phys.30. 2649-2656 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 伊藤 博康他: "マルチショット・パルスレ-ザ-蛍光顕微鏡による高速細胞現象のイメ-ジング" 計測自動制御学会論文集. 28. 156-157 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] K.kimura et al.: "Fluorescence quenching measurements of the membrane bound lipid haptens with different length spacers." Biochim.Biophys.Acta.

    • Related Report
      1991 Annual Research Report
  • [Publications] 木下 一彦,宝谷 紘一編: "限界を越える生物顕微鏡ー見えないものを見る" 学会出版センタ-, 177 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] K.kinosita,Jr.et al.: "Events of membrane electroporation visualized on a time scale from microsecond to seconds.(in Guide to Electroporation and Electrofusion)" Academic Press, 29-46 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] K.Kimura,et al.: "Location of MembraneーBound Hapten with Different Length Spacers" Immunol.69. 323-328 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] M.Hibino et al.: "Membrane Conductance of an Electropotated Cell Analyzed by Submicrosecond Imaging of Transmembrane Potential" Biophys.J.59. 209-220 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] H.Hyuga,et al.: "Deformation of Vesicles under the Influence of Strong Electric Fields I" Jpn.J.Appl.Phys. (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] H.Hyuga,et al.: "Deformation of Vesicles under the Influence of Strong Electric Fields II" Jpn.J.Appl.Phys.

    • Related Report
      1990 Annual Research Report
  • [Publications] A.NasudaーKouyama,et al.: "Effect of a lightinduced pH gradient on purpleーtoーbluecand purpleーtoーred transitions of bacteriorhodopsin" Biochemisty. 29. 6778-6788 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] H.Itoh,et al.: "MultiーShot Pulsed Laser Fluorescence Microscope System" TimeーResolved Laser Spectroscopy in Biochemistry II. 49-53 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] K.Kinosita,Jr.,et al.: "Submicrocecond Imaging under a PulsedーLaser Fluorescence Microscope.Electroporation of Cell Membrane Timeーand SpaceーResolved" KTK Sientific Publishers, (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] K.Kinosita,Jr.et al.: "Events of Membrane Electroporation Visualized on a Time Scale from Microsecond to Seconds" Academic Press, (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] T.Kouyama et al.: "Correlation between Internal Motion and Emission Kinetics of Tryptophan Residues in Proteins" Eur.J.Biochem.182. 517-521 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] S.Suzuki et al.: "Independent Flexible Motion of Submolecular Domains of the Ca^<2+>,Mg^<2+> ATPase of Sarcoplasmic Reticulum Measured by Time-resolved Fluores-cence Depolarization of Site-Specifically Attached Probes" Biochemistry. 28. 7734-7740 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] H.Itoh et al.: "Multi-Shot Pulsed Laser Fluorescence Microscope System" Proc.SPIE. 1204. (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] M.Shigemori et al.: "“Electroporation of Cell Membrane under a Pulsed Laser Fluorescence Microscope"in“Dynamic Behavior of Macromolecules,Colloids,Liquid Crystals,and Biological Systems by Optical and Electro-optical Methods"" Hirokawa Publishing Co., 311-316 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] K.Kinosita,Jr.,et al.: "“Submicrosecond Imaging under a Pulsed Laser Fluorescence Microscope.Electroporation of Cell Membrane Time- and Space-Resolved"in“Science on Form"(in press)" KTK Scientific Publishers, (1990)

    • Related Report
      1989 Annual Research Report

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Published: 1989-04-01   Modified: 2016-04-21  

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