Fluorescence Imaging Approaches in Flavivirus Research

Abstract

The genus Flavivirus within the family Flaviviridae contains many arthropod-borne infectious agents of medical relevance like dengue virus (DENV), Zika virus (ZIKV), and West Nile virus (WNV), among others, that cause epi- demics of hemorrhagic fevers and encephalitis for which there are no antiviral treatments and effective vaccines available. Fluorescence imaging is a powerful and versatile research tool for the study of fl aviviral diseases. This tool can be com- plemented with biochemical and molecular methods to gain insight into the mechanisms of fl avivirus infection and immunity, in order to develop feasible prophylactic and therapeutic interventions to lower their impact on public health. Here we discuss the basics of the fl uorescence imaging techniques currently employed in fl avivirus research, including immuno fl uorescence assay (IFA), fl uorescence in situ hybridization (FISH), fl uorescence-labeled viral particles, fl uorescent labeling of cytopathic effect (CPE), subgenomic reporter replicons (SRRs)/reporter virus particles (RVPs), and cell-based molecular reporters (CBMRs). We also address the advantages of each application based on our own experience and included some of our protocols to facilitate its applications.