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http://hdl.handle.net/1942/27258
Title: | Single Viruses on the Fluorescence Microscope: Imaging Molecular Mobility, Interactions and Structure Sheds New Light on Viral Replication | Authors: | Parveen, Nagma Borrenberghs, Doortje Rocha, Susana HENDRIX, Jelle |
Issue Date: | 2018 | Source: | Viruses, 10(5), p. 1-21 (Art N° 250) | Abstract: | Viruses are simple agents exhibiting complex reproductive mechanisms. Decades of research have provided crucial basic insights, antiviral medication and moderately successful gene therapy trials. The most infectious viral particle is, however, not always the most abundant one in a population, questioning the utility of classic ensemble-averaging virology. Indeed, viral replication is often not particularly efficient, prone to errors or containing parallel routes. Here, we review different single-molecule sensitive fluorescence methods that we employ routinely to investigate viruses. We provide a brief overview of the microscopy hardware needed and discuss the different methods and their application. In particular, we review how we applied (i) single-molecule Förster resonance energy transfer (smFRET) to probe the subviral human immunodeficiency virus (HIV-1) integrase (IN) quaternary structure; (ii) single particle tracking to study interactions of the simian virus 40 with membranes; (iii) 3D confocal microscopy and smFRET to quantify the HIV-1 pre-integration complex content and quaternary structure; (iv) image correlation spectroscopy to quantify the cytosolic HIV-1 Gag assembly, and finally; (v) super-resolution microscopy to characterize the interaction of HIV-1 with tetherin during assembly. We hope this review is an incentive for setting up and applying similar single-virus imaging studies in daily virology practice | Notes: | Hendrix, J (reprint author), Katholieke Univ Leuven, Dept Chem, Mol Imaging & Photon Div, Lab Photochem & Spect, B-3001 Leuven, Belgium nagma.parveen@kuleuven.be; doortjeborrenberghs@gmail.com; susana.rocha@kuleuven.be; jelle.hendrix@uhasselt.be | Keywords: | wide-field fluorescence microscopy; confocal laser scanning microscopy; super-resolution microscopy; single virus imaging; raster image correlation spectroscopy; single particle tracking; Förster resonance energy transfer; HIV; simian virus 40, oligomerization; stoichiometry | Document URI: | http://hdl.handle.net/1942/27258 | e-ISSN: | 1999-4915 | DOI: | 10.3390/v10050250 | ISI #: | 000435198000035 | Rights: | © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). | Category: | A1 | Type: | Journal Contribution | Validations: | ecoom 2019 |
Appears in Collections: | Research publications |
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File | Description | Size | Format | |
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viruses-10-00250.pdf | Published version | 2.8 MB | Adobe PDF | View/Open |
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