title: Infection Counter: Automated Quantification of in Vitro Virus Replication by Fluorescence Microscopy.
abstract: The ability to accurately and reliably quantify viral infection is essential to basic and translational virology research. Here, we describe a simple and robust automated method for using fluorescence microscopy to estimate the proportion of virally infected cells in a monolayer. We provide details of the automated analysis workflow along with a freely available open-source ImageJ plugin, Infection Counter, for performing image quantification. Using hepatitis C virus (HCV) as an example, we have experimentally verified our method, demonstrating that it is equivalent, if not better, than the established focus-forming assay. Finally, we used Infection Counter to assess the anti-HCV activity of SMBz-CsA, a non-immunosuppressive cyclosporine analogue.
authors.affiliation: MRC Laboratory for Molecular Cell Biology, University College London, London WC1E 6BT, UK. s.culley@ucl.ac.uk.|Department of Cell and Developmental Biology, University College London, London WC1E 6BT, UK. s.culley@ucl.ac.uk.
authors.forename: Siân
authors.initials: S
authors.lastname: Culley
authors.affiliation: Division of Infection and Immunity, University College London, London WC1E 6BT, UK. g.towers@ucl.ac.uk.
authors.forename: Greg J
authors.initials: GJ
authors.lastname: Towers
authors.affiliation: Wolfson Institute for Biomedical Research, University College London, London WC1E 6BT, UK. d.selwood@ucl.ac.uk.
authors.forename: David L
authors.initials: DL
authors.lastname: Selwood
authors.affiliation: MRC Laboratory for Molecular Cell Biology, University College London, London WC1E 6BT, UK. r.henriques@ucl.ac.uk.|Department of Cell and Developmental Biology, University College London, London WC1E 6BT, UK. r.henriques@ucl.ac.uk.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Henriques
authors.affiliation: Division of Infection and Immunity, University College London, London WC1E 6BT, UK. j.grove@ucl.ac.uk.
authors.forename: Joe
authors.initials: J
authors.lastname: Grove
mesh_terms: D057205:Automation, Laboratory / Q000379:methods*; D016174:Hepacivirus / Q000502:physiology*; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D014773:Virology / Q000379:methods*; D014779:Virus Replication
keywords: ImageJ; focus-forming assay; hepatitis C virus; image quantification; plaque assay