Ex Vivo Human Neutrophil Swarming Against Live Microbial Targets

Neutrophils often communicate with each other and coordinate their actions to seal off large sites of injury and infection that individual neutrophils could not cover. The concerted actions of neutrophils are essential for the expeditious protection of healthy tissues from wounds and microbes. These processes, collectively known as swarming, are typically studied in vivo in mice. However, these studies are low throughput and their relevance to human disease is limited. Recently, new tools have been developed for the study of human neutrophil swarming ex vivo. The emergent microscale swarming assays are providing significant insights into the molecular mediators of swarming. By enabling the direct study of human cells, these assays also shed new light on human diseases and host responses against infections. Here, we describe a robust technique for manufacturing microscale swarming arrays with live microbial targets (e.g., clusters of Candida albicans). These arrays allow for the direct, precise, and efficient interrogation of the antimicrobial functions of human swarming against a variety of targets.