Infection Counter: Automated Quantification of in Vitro Virus Replication by Fluorescence Microscopy

• Published in: Viruses Vol. 8; no. 7; p. 201
• Main Authors: Culley, Siân, Towers, Greg J, Selwood, David L, Henriques, Ricardo, Grove, Joe
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI 21-07-2016; MDPI AG
• Subjects: Automation, Laboratory - methods; focus-forming assay; Hepacivirus - physiology; hepatitis C virus; Humans; image quantification; ImageJ; Microscopy, Fluorescence - methods; plaque assay; Virology - methods; Virus Replication; ImageJ; hepatitis C virus; focus-forming assay; image quantification; plaque assay
• ISSN: 1999-4915; 1999-4915
• DOI: 10.3390/v8070201

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.