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

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 elec...

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Detalles Bibliográficos
Autores principales: Gray, Robert D. M., Mercer, Jason, Henriques, Ricardo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MyJove Corporation 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5564481/
https://www.ncbi.nlm.nih.gov/pubmed/28448005
http://dx.doi.org/10.3791/55471
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author Gray, Robert D. M.
Mercer, Jason
Henriques, Ricardo
author_facet Gray, Robert D. M.
Mercer, Jason
Henriques, Ricardo
author_sort Gray, Robert D. M.
collection PubMed
description 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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spelling pubmed-55644812017-09-01 Open-source Single-particle Analysis for Super-resolution Microscopy with VirusMapper Gray, Robert D. M. Mercer, Jason Henriques, Ricardo J Vis Exp Molecular Biology 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. MyJove Corporation 2017-04-09 /pmc/articles/PMC5564481/ /pubmed/28448005 http://dx.doi.org/10.3791/55471 Text en Copyright © 2017, Journal of Visualized Experiments http://creativecommons.org/licenses/by/3.0/us/ This is an open-access article distributed under the terms of the Creative Commons Attribution 3.0 License. To view a copy of this license, visithttp://creativecommons.org/licenses/by/3.0/us/
spellingShingle Molecular Biology
Gray, Robert D. M.
Mercer, Jason
Henriques, Ricardo
Open-source Single-particle Analysis for Super-resolution Microscopy with VirusMapper
title Open-source Single-particle Analysis for Super-resolution Microscopy with VirusMapper
title_full Open-source Single-particle Analysis for Super-resolution Microscopy with VirusMapper
title_fullStr Open-source Single-particle Analysis for Super-resolution Microscopy with VirusMapper
title_full_unstemmed Open-source Single-particle Analysis for Super-resolution Microscopy with VirusMapper
title_short Open-source Single-particle Analysis for Super-resolution Microscopy with VirusMapper
title_sort open-source single-particle analysis for super-resolution microscopy with virusmapper
topic Molecular Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5564481/
https://www.ncbi.nlm.nih.gov/pubmed/28448005
http://dx.doi.org/10.3791/55471
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