Local and global analysis of endocytic patch dynamics in fission yeast using a new "temporal superresolution" realignment method
Local and global analysis of endocytic patch dynamics in fission yeast using a new "temporal superresolution" realignment method
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DOI：
10.1091/mbc.e13-01-0004
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发表时间：
2014-11-05
影响因子：
3.3
通讯作者：
Pollard, Thomas D.
中科院分区：
文献类型：
--
作者：
Berro, Julien;Pollard, Thomas D.
定量显微镜是一种有价值的工具，用于推断细胞过程的分子机制，如网格蛋白介导的内吞作用，但定量显微镜，以达到其潜力，数据收集和分析需要改进。我们引入了新的工具来跟踪和计数内吞补丁裂变酵母，以提高质量的定量显微镜电影提取的数据。我们提出了一种通用的方法来实现“时间超分辨率”对齐时间数据集具有更高的时间分辨率比测量间隔。这些方法使我们能够从fimbrin-mEGFP的荧光测量中提取有关野生型细胞中内吞肌动蛋白斑块的新信息。我们表明，肌动蛋白的组装和拆卸的时间过程不同，
Quantitative microscopy is a valuable tool for inferring molecular mechanisms of cellular processes such as clathrin-mediated endocytosis, but, for quantitative microscopy to reach its potential, both data collection and analysis needed improvement. We introduce new tools to track and count endocytic patches in fission yeast to increase the quality of the data extracted from quantitative microscopy movies. We present a universal method to achieve "temporal superresolution" by aligning temporal data sets with higher temporal resolution than the measurement intervals. These methods allowed us to extract new information about endocytic actin patches in wild-type cells from measurements of the fluorescence of fimbrin-mEGFP. We show that the time course of actin assembly and disassembly varies