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1991 Fiscal Year Final Research Report Summary

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
KeywordsMulti-shot pulsed laser microscope / Multiview microscopy / Fluorescence polarization imaging / Cell eletroporation / Cell celctrodeformation / Actomyosin motor / Molecular mechanism of fertilization
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.

  • Research Products

    (38 results)

All Other

All Publications (38 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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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

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Published: 1999-03-16  

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