Open-source single-particle analysis for super-resolution microscopy with virusmapper

Research output: Contribution to journalArticlepeer-review

Authors

Colleges, School and Institutes

External organisations

  • UCL

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.

Details

Original languageEnglish
Article numbere55471
JournalJournal of Visualized Experiments
Volume2017
Issue number122
Publication statusPublished - 9 Apr 2017

Keywords

  • Fiji, Fluorescence microscopy, Image analysis, Issue 122, Molecular Biology, Single-particle analysis, Software, Super-resolution microscopy, Vaccinia, Viruses