| PC12活细胞中单个分泌囊泡的动态成像 |
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| 引用本文： | 吴政星,夏胜,许亮,白丽,徐涛.PC12活细胞中单个分泌囊泡的动态成像[J].生物化学与生物物理学报,2003,35(4):381-386. |
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| 作者姓名： | 吴政星 夏胜 许亮 白丽 徐涛 |
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| 作者单位： | 华中科技大学生命科学与技术学院生物物理与生物化学研究所 武汉430074
(吴政星，夏胜，许亮，白丽)，华中科技大学生命科学与技术学院生物物理与生物化学研究所 武汉430074(徐涛) |
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| 基金项目： | 国家自然科学基金资助项目 (No .3 0 0 2 5 0 2 3 ,No.3 0 13 0 2 3 0 )，国家重点基础研究发展规划 (973计划 )项目 (No .G19990 5 40 0 0 )~~ |
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| 摘 要： | 囊泡的荧光标记和动态显微成像观察是研究蛋白质和膜转运机制的重要手段。采用EGFP hpNPY融合荧光蛋白标记PC12细胞的致密大囊泡 ,用全内反射和宽场荧光显微镜对PC12细胞进行成像研究。结果发现 :普通的宽场荧光成像模糊不清 ,难以观察到单个囊泡 ;而全内反射荧光成像则可清晰地分辨出呈现为离散荧光点的单个囊泡 ;并且进一步利用全内反射荧光成像直接观察到了活的PC12细胞中单个囊泡的转运、锚定及与细胞膜的融合过程 ,证实了囊泡的锚定过程是可逆的。
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| 关 键 词： | 全内反射荧光显微镜 绿色荧光蛋白 囊泡 PC12细胞 胞吐 |
Dynamic Imaging of Single Secretory Granule in Cultured PC12 Cells |
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| WU Zheng-Xing,XIA Sheng,XU Liang,BAI Li,XU Tao.Dynamic Imaging of Single Secretory Granule in Cultured PC12 Cells[J].Acta Biochimica et Biophysica Sinica,2003,35(4):381-386. |
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| Authors: | WU Zheng-Xing XIA Sheng XU Liang BAI Li XU Tao |
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| Institution: | WU Zheng-Xing,XIA Sheng,XU Liang,BAI Li,XU Tao * |
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| Abstract: | Fluorescent labeling and dynamic imaging of secretory vesicles are new powerful means to study the mechanisms of protein and membrane trafficking. The large dense-core granules in PC12 cells were labeled by transfecting EGFP-hpNPY chimera and imaged with epi-fluorescence and evanescent field (EF) fluorescence simultaneously. Under epi-fluorescence illumination, the cells exhibited nearly uniform fluorescence, however, EF-fluorescence excitation revealed distinct fluorescent spots corresponding to GFP-labeled granules. The trafficking, docking and fusing with plasma membrane of individual granule in cultured PC12 cells were observed directly by EF fluorescence imaging. |
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| Keywords: | TIRFM GFP granule PC12 cells exocytosis |
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