Polarized Fluorescence Microscopy to Study Cytoskeleton Assembly and Organization in Live Cells.
Polarized Fluorescence Microscopy to Study Cytoskeleton Assembly and Organization in Live Cells.
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DOI：
10.1002/0471143030.cb0429s67
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发表时间：
2015-06-01
影响因子：
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通讯作者：
Gladfelter, Amy S
中科院分区：
文献类型：
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作者：
McQuilken, Molly;Mehta, Shalin B;Gladfelter, Amy S
测量活细胞中分子的位置和组织对于理解不同的生物过程至关重要。偏振光显微镜提供了一种非破坏性的手段来评估亚细胞结构域内的秩序。当与荧光显微镜和GFP标记的蛋白质相结合时，这种方法可以揭示特定分子群体中的组织。本单元描述了使用偏振荧光显微镜和OpenPolScope开放获取软件（http：//www.example.com）测量细胞骨架组分的结构动力学的方案。www.openpolscope.org该方案描述了线性偏振器或液晶（LC）通用补偿器的安装，系统的校准，偏振荧光成像和分析。使用OpenPolScope软件和硬件可以进行可靠、用户友好的图像采集，以测量和分析偏振荧光。
The measurement of not only the location but also the organization of molecules in live cells is crucial to understanding diverse biological processes. Polarized light microscopy provides a nondestructive means to evaluate order within subcellular domains. When combined with fluorescence microscopy and GFP-tagged proteins, the approach can reveal organization within specific populations of molecules. This unit describes a protocol for measuring the architectural dynamics of cytoskeletal components using polarized fluorescence microscopy and OpenPolScope open-access software (http://www.openpolscope.org). The protocol describes installation of linear polarizers or a liquid crystal (LC) universal compensator, calibration of the system, polarized fluorescence imaging, and analysis. The use of OpenPolScope software and hardware allows for reliable, user-friendly image acquisition to measure and analyze polarized fluorescence.