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Quantification and Visualization of Neutrophil Extracellular Traps (NETs) from Murine Bone Marrow-Derived Neutrophils

  • Linda Vong
  • Philip M. Sherman
  • Michael GlogauerEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1960)

Abstract

Neutrophils are some of the first leukocytes to respond to inflammatory stimuli. Once recruited, these cells are equipped with an assortment of proteolytic enzymes and antimicrobial factors that disarm and degrade pathogens. Neutrophils employ a highly novel mechanism to contain and trap bacteria in the local inflammatory microenvironment, termed neutrophil extracellular traps (NETs). During NET formation, neutrophils eject weblike structures of chromatin, which captures and immobilizes invading pathogens. In this chapter, we describe protocols to isolate bone marrow-derived neutrophils from mice. We further describe in vitro methods to spectrophotometrically quantify, immunolabel, and visualize NET structures.

Keywords

Neutrophil extracellular trap Bone marrow neutrophil Nucleic acid stain Histone H3 Elastase Fluorescence microscopy 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Linda Vong
    • 1
  • Philip M. Sherman
    • 1
    • 2
  • Michael Glogauer
    • 2
    Email author
  1. 1.Cell Biology Program, Research Institute, Hospital for Sick ChildrenUniversity of TorontoTorontoCanada
  2. 2.Faculty of DentistryUniversity of TorontoTorontoCanada

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