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

Gray, RDM; Mercer, J; Henriques, R; (2017) Open-source Single-particle Analysis for Super-resolution Microscopy with VirusMapper. J Vis Exp , 2017 (122) , Article e55471. 10.3791/55471. Green open access

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Abstract

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.

Type: Article
Title: Open-source Single-particle Analysis for Super-resolution Microscopy with VirusMapper.
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.3791/55471
Publisher version: http://dx.doi.org/10.3791/55471
Language: English
Additional information: Copyright © 2017 Creative Commons Attribution 3.0 License
Keywords: Molecular Biology, super-resolution microscopy, fluorescence microscopy, image analysis, single-particle analysis, software, viruses, Fiji, vaccinia
UCL classification: UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Cell and Developmental Biology
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Computer Science
URI: http://discovery.ucl.ac.uk/id/eprint/1554755
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