Flow virometry: recent advancements, best practices, and future frontiers

The imperative for developing robust tools to detect, analyze, and characterize viruses has become increasingly evident as they continue to threaten human health. In this review, we focus on recent advancements in studying human viruses with flow virometry (FV), an emerging technique that has gained considerable momentum over the past 5 years. These advancements include the application of FV in viral surface phenotyping, viral protein functionality, virus sorting, vaccine development, and diagnostics. With examples illustrated using primary data from our recent studies, we demonstrate that FV is a powerful yet underutilized methodology that, when employed with best practices and experimental rigor, can be highly valuable for studying individual virion heterogeneity, virus phenotypes, and virus-antibody interactions. In this review, we also address the current challenges when performing FV studies, propose strategies to overcome these obstacles, and outline best practices for both new and experienced researchers. Finally, we discuss the promising future prospects of FV within the broader context of virology research.