Open-source Single-particle Analysis for Super-resolution Microscopy with VirusMapper.

Open-source Single-particle Analysis for Super-resolution Microscopy with VirusMapper.
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使用 VirusMapper 进行超分辨率显微镜的开源单粒子分析。

DOI:
10.3791/55471
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发表时间:
2017-04-09
期刊:
Journal of visualized experiments : JoVE
影响因子:
--
通讯作者:
Henriques R
Henriques R
中科院分区:
其他
文献类型:
--
作者:
Gray RDM;Mercer J;Henriques R

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超分辨率荧光显微镜目前正在彻底改变细胞生物学的研究。其突破300纳米分辨率限制的能力使得纳米级生物复合物和过程的常规成像成为可能。分辨率的提高也意味着在电子显微镜中流行的方法,如单粒子分析,可以很容易地应用于超分辨率荧光显微镜。通过将这种分析方法与超分辨率光学成像相结合,利用荧光显微镜的分子特异性标记能力来生成亚稳结构内分子元素的结构图成为可能。为此,我们开发了一种新的算法——VirusMapper,它被打包成一个易于使用、高性能和高吞吐量的ImageJ插件。本文介绍了该软件的深入指南,展示了其揭示生物分子复合物中新结构特征的能力。在这里,我们介绍了如何组装兼容的数据,并提供了一个循序渐进的协议,如何使用该算法将单粒子分析应用于超分辨率图像。
Super-resolution fluorescence microscopy is currently revolutionizing cell biology research. Its capacity to break the resolution limit of around 300 nm allows for the routine imaging of nanoscale biological complexes and processes. This increase in resolution also means that methods popular in electron microscopy, such as single-particle analysis, can readily be applied to super-resolution fluorescence microscopy. By combining this analytical approach with super-resolution optical imaging, it becomes possible to take advantage of the molecule-specific labeling capacity of fluorescence microscopy to generate structural maps of molecular elements within a metastable structure. To this end, we have developed a novel algorithm — VirusMapper — packaged as an easy-to-use, high-performance, and high-throughput ImageJ plugin. This article presents an in-depth guide to this software, showcasing its ability to uncover novel structural features in biological molecular complexes. Here, we present how to assemble compatible data and provide a step-by-step protocol on how to use this algorithm to apply single-particle analysis to super-resolution images.
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