Microfluidic Platform to Quantify Neutrophil Migratory Decision-Making

: Neutrophils are the most abundant leukocytes in blood, serving as the first line of host defense in tissue damage and infections. Upon activation by chemokines released from pathogens or injured tissues, neutrophils migrate through complex tissue microenvironments toward sites of infections along the chemokine gradients, in a process named chemotaxis. However, current methods for measuring neutrophil chemotaxis require large volumes of blood and are often bulk, endpoint measurements. To address the need for rapid and robust assays, we engineered a novel dual gradient microfluidic platform that precisely quantifies neutrophil migratory decision-making with high temporal resolution. Here, we present a protocol to measure neutrophil migratory phenotypes (velocity, directionality) with single-cell resolution.