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

Authors: Robert DM Gray, Jason Mercer, Ricardo Henriques
Technologies: NanoJ-VirusMapper
Paper published in JoVE, January 2017

Open-source single-particle analysis for super-resolution microscopy with VirusMapper
DOI: 10.3791/55471

The manuscript by Gray, Mercer, and Henriques published in the Journal of Visualized Experiments in 2017 describes the development and application of VirusMapper, an open-source software package for single-particle analysis (SPA) of super-resolution fluorescence microscopy images. Super-resolution microscopy has revolutionized cell biology research by enabling the imaging of nanoscale biological complexes and processes with molecular specific labeling. SPA, a method commonly used in electron microscopy, can be applied to super-resolution data to improve resolution, precision, or signal-to-noise. However, the lack of available software for combining super-resolution and SPA has hindered their routine use together. VirusMapper is a plugin for the popular image processing software ImageJ/Fiji designed to provide fast, user-friendly, multi-channel naïve averaging of structures imaged with SR microscopy. It can be used to produce high-precision molecular models of any known structure, allowing for the calculation of average dimensions and other parameters. The software is particularly useful for separating populations of structures, providing for the determination of distinct orientations or different morphological states. It can also be employed for reference-based structure discovery by using a known reference channel. The manuscript provides a detailed guide to using VirusMapper, including instructions for downloading and installing the software, as well as example data for practicing.