Infection counter: Automated quantification of in vitro virus replication by fluorescence microscopy

TY - JOUR

T1 - Infection counter

T2 - Automated quantification of in vitro virus replication by fluorescence microscopy

AU - Culley, Siân

AU - Towers, Greg J.

AU - Selwood, David L.

AU - Henriques, Ricardo

AU - Grove, Joe

N1 - Funding Information: We would like to thank Lucas Walker for critical reading of the manuscript. Joe Grove is supported by a Sir Henry Dale Fellowship jointly funded the Wellcome Trust and Royal Society (Grant Number 107653/Z/15/Z). Si?n Culley and Ricardo Henriques are supported by grant MR/K015826/1 from the Medical Research Council and grant BB/M022374/1 from the Biotechnology and Biological Sciences Research Council. Publisher Copyright: © 2016 by the authors; licensee MDPI, Basel, Switzerland.

PY - 2016/7/21

Y1 - 2016/7/21

N2 - 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.

AB - 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.

KW - Focus-forming assay

KW - Hepatitis C virus

KW - Image quantification

KW - ImageJ

KW - Plaque assay

UR - http://www.scopus.com/inward/record.url?scp=84979224962&partnerID=8YFLogxK

U2 - 10.3390/v8070201

DO - 10.3390/v8070201

M3 - Article

C2 - 27455304

AN - SCOPUS:84979224962

SN - 1999-4915

VL - 8

JO - Viruses

JF - Viruses

IS - 7

M1 - 201

ER -