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Related Experiment Video

Updated: Mar 12, 2026

Morphological and Compositional Analysis of Neutrophil Extracellular Traps Induced by Microbial and Chemical Stimuli
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Induction and Quantification of NETosis.

Carmelo Carmona-Rivera1, Mariana J Kaplan1

  • 1Systemic Autoimmunity Branch, Intramural Research Program, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, Maryland.

Current Protocols in Immunology
|November 2, 2016
PubMed
Summary

Neutrophil extracellular traps (NETs) are web-like structures released by neutrophils. Standardized methods are presented for inducing and quantifying NETs to improve research consistency in health and disease.

Keywords:
flow cytometryimmunofluorescenceneutrophil extracellular trapsneutrophils

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Area of Science:

  • Immunology
  • Cell Biology
  • Biochemistry

Background:

  • Neutrophil extracellular traps (NETs) are crucial in host defense but also implicated in inflammatory diseases.
  • NETosis, the process of NET extrusion, requires standardized induction and quantification methods for reliable research.
  • Current methodologies lack consistency, hindering comparative studies and understanding of NETs' role in pathology.

Purpose of the Study:

  • To present validated methods for isolating human neutrophils.
  • To describe standardized protocols for inducing NETs using various stimuli.
  • To offer reliable low- and high-throughput methods for NET quantification.

Main Methods:

  • Isolation of human neutrophils from peripheral blood.
  • Induction of NETs via stimuli targeting NADPH oxidase or mitochondrial reactive oxygen species production.
  • Low- and high-throughput quantification assays for NETs.

Main Results:

  • Established protocols for reproducible NET induction and isolation of neutrophils.
  • Demonstrated efficacy of ROS-dependent pathways in NET formation.
  • Provided adaptable quantification techniques for diverse research needs.

Conclusions:

  • Standardized methods are essential for advancing NET research.
  • The presented protocols facilitate consistent and comparable results in NETosis studies.
  • Accurate NET quantification aids in understanding their implications in various diseases.