Protocol to investigate the role of NETosis in health and disease via a high-throughput neutrophil-like model using quantitative imaging.

Neutrophil extracellular traps (NETs) are a crucial host-defense mechanism employed by neutrophils in response to various aggregates and contribute to pathology in diverse diseases, from chronic inflammation to sepsis. Here, we present a protocol to investigate the role of NETosis, the release of NETs, in health and disease via a high-throughput neutrophil-like model using quantitative imaging. We describe steps for preparing stimulated, differentiated Human Leukemia-60 (HL-60) cells; immunostaining; high-throughput microscopy; and NETosis quantification using CellProfiler.