@article {404661,
	title = {A high-throughput drop microfluidic system for virus culture and analysis},
	journal = {Journal of Virological Methods},
	volume = {213},
	number = {1},
	year = {2015},
	pages = {111-117},
	abstract = {
	High mutation rates and short replication times lead to rapid evolution in RNA viruses. New tools for high-throughput culture and analysis of viral phenotypes will enable more effective studies of viral evolutionary processes. A water-in-oil drop microfluidic system to study virus{\textendash}cell interactions at the single event level on a massively parallel scale is described here. Murine norovirus (MNV-1) particles were co-encapsulated with individual RAW 264.7 cells in 65 pL aqueous drops formed by flow focusing in 50 μm microchannels. At low multiplicity of infection (MOI), viral titers increased greatly, reaching a maximum 18 h post-encapsulation. This system was employed to evaluate MNV-1 escape from a neutralizing monoclonal antibody (clone A6.2). Further, the system was validated as a means for testing escape from antibody neutralization using a series of viral point mutants. Finally, the replicative capacity of single viral particles in drops under antibody stress was tested. Under standard conditions, many RNA virus stocks harbor minority populations of genotypic and phenotypic variants, resulting in quasispecies. These data show that when single cells are encapsulated with single viral particles under antibody stress without competition from other virions, the number of resulting infectious particles is nearly equivalent to the number of viral genomes present. These findings suggest that lower fitness virions can infect cells successfully and replicate, indicating that the microfluidics system may serve as an effective tool for isolating mutants that escape evolutionary stressors.

},
	keywords = {Microfluidics Murine norovirus Antibody neutralization Evolution},
	url = {https://www.sciencedirect.com/science/article/abs/pii/S0166093414004662?via\%3Dihub},
	author = {Fischer, Audrey E. and Wu, Susan K. and Proescher, Jody B. G. and Rotem, Assaf and Chang, Connie B. and Zhang, Huidan and Tao, Ye and Mehoke, Thomas S. and Thielen, Peter M. and Kolawole, Abimbola O. and Smith, Thomas J. and Wobus, Christiane E. and Weitz, David A and Lin, Jeffrey S. and Feldman, Andrew B. and Wolfe, Joshua T.}
}
