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| 双光子激发荧光各向异性度的成像 | |
| 引用本文： | 汪雪峰,王 毅,蒋 艳,马 辉.双光子激发荧光各向异性度的成像[J].生物化学与生物物理进展,2005,32(2):161-167. |
| 作者姓名： | 汪雪峰 王 毅 蒋 艳 马 辉 |
| 作者单位： | 1. 清华大学物理系,原子分子纳米科学教育部重点实验室,北京,100084 2. 北京中医药大学病理室,北京,100029 3. 清华大学物理系,原子分子纳米科学教育部重点实验室,北京,100084;清华大学深圳研究生院,深圳,518057 |
| 基金项目： | 国家自然科学基金海外青年学者合作基金(19928408), 国家自然科学重点基金(60138010)和面上基金(10274039)资助项目. |
| 摘 要： | 荧光各向异性度 (fluorescence anisotropy) 测量可以获得荧光分子的转动速度信息，进而了解分子质量、结构、以及与周边环境的相互作用情况 . 围绕一台双光子激发扫描荧光成像系统，通过改变外光路和图像记录与处理程序，从而实现了双光子激发荧光各向异性度成像，并针对一些典型样品和体系，展示了该方法的应用 . 实验中观察了 FITC 荧光分子、 FITC 结合的 CD44 抗体分子及与肿瘤细胞表面受体结合的 FITC-CD44 抗体分子 . 测量结果表明，不同分子质量、不同微观环境状态下的荧光分子，其各向异性度大小不同，在各向异性度图中能够被明显区分 . 荧光各向异性度成像能够定量测量样品微区的各向异性度值，并以二维图像的形式直观表达，是各向异性度测量与成像技术的良好结合 . |
| 关 键 词： | 荧光各向异性度，双光子扫描成像，细胞膜流动性，小檗碱，肿瘤细胞 |
Two-photon Fluorescence Anisotropy Imaging |
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| WANG Xue-Feng,WANG Yi,JIANG Yan and MA Hui.Two-photon Fluorescence Anisotropy Imaging[J].Progress In Biochemistry and Biophysics,2005,32(2):161-167. | |
| Authors: | WANG Xue-Feng WANG Yi JIANG Yan and MA Hui |
| Institution: | The Key Laboratory of Atomic and Molecular Nanosciences, Ministry of Education, Department of Physics, Tsinghua University, Beijing 100084, China;The Key Laboratory of Atomic and Molecular Nanosciences, Ministry of Education, Department of Physics, Tsinghua University, Beijing 100084, China;Graduate School at Shenzhen, Tsinghua University, Shenzhen 518057, China;1)The Key Laboratory of Atomic and Molecular Nanosciences, Ministry of Education, Department of Physics, Tsinghua University, Beijing 100084, China; 2)Graduate School at Shenzhen, Tsinghua University, Shenzhen 518057, China |
| Abstract: | A new method is developed for two-photon fluorescence anisotropy imaging. Its biological applications are tested. This system is based on a two-photon laser scanning fluorescence microscope. A polarization beam splitter was inserted into the optical path to separated fluorescence into components of orthogonal polarization. By rotating the polarization of the excitation beam by 90 ° , four images were collected for each sample. These images were then processed pixel-by-pixel to generate a new fluorescence anisotropy image. The capability of this method is tested for different samples, including FITC, FITC-CD44 in solution and FITC-CD44 attached to the membranes of tumor cells. The results show that fluorescence images can distinguish fluorescence molecules of different molecular mass, or detect changes in microenvironment. |
| Keywords: | fluorescence anisotropy two-photon scanning microscopy fluidity of membrane berberine tumor cell |
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