title: Labeling of virus components for advanced, quantitative imaging analyses.
abstract: In recent years, investigation of virus-cell interactions has moved from ensemble measurements to imaging analyses at the single-particle level. Advanced fluorescence microscopy techniques provide single-molecule sensitivity and subdiffraction spatial resolution, allowing observation of subviral details and individual replication events to obtain detailed quantitative information. To exploit the full potential of these techniques, virologists need to employ novel labeling strategies, taking into account specific constraints imposed by viruses, as well as unique requirements of microscopic methods. Here, we compare strengths and limitations of various labeling methods, exemplify virological questions that were successfully addressed, and discuss challenges and future potential of novel approaches in virus imaging.
authors.affiliation: Department of Infectious Diseases, Virology, University Hospital Heidelberg, Germany.
authors.forename: Volkan
authors.initials: V
authors.lastname: Sakin
authors.affiliation: Structural and Computational Biology Unit, Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
authors.forename: Giulia
authors.initials: G
authors.lastname: Paci
authors.affiliation: Structural and Computational Biology Unit, Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
authors.forename: Edward A
authors.initials: EA
authors.lastname: Lemke
authors.affiliation: Department of Infectious Diseases, Virology, University Hospital Heidelberg, Germany.
authors.forename: Barbara
authors.identifier: 0000-0001-5726-5585
authors.initials: B
authors.lastname: Müller
mesh_terms: D005456:Fluorescent Dyes / Q000378:metabolism; D021621:Imaging, Three-Dimensional; D013194:Staining and Labeling; D014779:Virus Replication; D014780:Viruses / Q000378:metabolism*
chemical_list: D005456:Fluorescent Dyes
keywords: amber suppression; click labeling; fluorescence; human immunodeficiency virus; imaging; nanoscopy; single-molecule techniques; virus