Quantification of Neturophil Extracellular Traps

Neutrophils are one of the first cells to respond to an inflammatory stimulus, and are equipped with an assortment of antimicrobial and proteolytic enzymes to disarm and degrade bacterial pathogens. A novel mechanism of bacterial trapping, termed neutrophil extracellular traps (NETs), was recently described whereby neutrophils were shown to cast out web-like structures of chromatin, capturing and immobilizing invading pathogens. Herein we describe protocols to isolate murine bone marrow-derived neutrophils,and spectrophotometrically quantify, immunolabel, and visualize NET structures in vitro.