| Project/Area Number |
01440095
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
生物物性学
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| Research Institution | Keio University |
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Principal Investigator |
KINOSHITA Kazuhiko Keio University, Faculty of Science and Technology, Professor, 理工学部, 教授 (30124366)
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| 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)
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| Project Period (FY) |
1989 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| 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)
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| Keywords | Multi-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 |
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| 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.
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