Multiple signal classification algorithm for super-resolution fluorescence microscopy.

Multiple signal classification algorithm for super-resolution fluorescence microscopy.
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超分辨率荧光显微镜的多个信号分类算法。

DOI:
10.1038/ncomms13752
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发表时间:
2016-12-09
影响因子:
16.6
通讯作者:
--
中科院分区:
综合性期刊1区
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--
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单分子定位技术受到长的采集和计算时间,或需要特殊的荧光团或生物毒性光化学环境的限制。在这里,我们提出了一种统计超分辨率技术的宽场荧光显微镜,我们称之为多信号分类算法,它有几个优点。它提供的分辨率低至至少50 nm,需要更少的帧和更低的激发功率,即使在高荧光团浓度下也能工作。此外,它与在记录的时间尺度上表现出闪烁的任何荧光团一起工作。与单分子定位技术和四种当代统计超分辨率方法相比,多信号分类算法显示出相当或更好的性能,用于体外肌动蛋白丝和其他独立获得的实验数据集的实验。我们还证明了超分辨率在245毫秒的时间尺度(使用49帧,每秒200帧)的活细胞微管和活细胞肌动蛋白丝成像没有成像缓冲液的样品。 单分子定位显微镜提供了超分辨率成像,但需要很长的采集时间和有毒的光化学环境。在这里,作者展示了一种多信号分类算法,该算法在生物有益环境中仅用50帧就实现了50 nm的分辨率。
Single-molecule localization techniques are restricted by long acquisition and computational times, or the need of special fluorophores or biologically toxic photochemical environments. Here we propose a statistical super-resolution technique of wide-field fluorescence microscopy we call the multiple signal classification algorithm which has several advantages. It provides resolution down to at least 50 nm, requires fewer frames and lower excitation power and works even at high fluorophore concentrations. Further, it works with any fluorophore that exhibits blinking on the timescale of the recording. The multiple signal classification algorithm shows comparable or better performance in comparison with single-molecule localization techniques and four contemporary statistical super-resolution methods for experiments of in vitro actin filaments and other independently acquired experimental data sets. We also demonstrate super-resolution at timescales of 245 ms (using 49 frames acquired at 200 frames per second) in samples of live-cell microtubules and live-cell actin filaments imaged without imaging buffers. Single-molecule localization microscopy offers super-resolution imaging, but needs a long acquisition time and a toxic photochemical environment. Here, the authors demonstrate a multiple signal classification algorithm that achieves a resolution of 50 nm with as few as 50 frames in a biologically conducive environment.
DOI: 10.1038/nmeth.1768
发表时间: 2011-11-06
期刊: NATURE METHODS
影响因子: 48
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发表时间: 2009-12-29
影响因子: 11.1
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影响因子: 7.8
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