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Related Experiment Video

Updated: May 15, 2026

Automated Image-Based Quantification of Neutrophil Extracellular Traps Using NETQUANT
07:33

Automated Image-Based Quantification of Neutrophil Extracellular Traps Using NETQUANT

Published on: November 27, 2019

Automatic quantification of in vitro NET formation.

Volker Brinkmann1, Christian Goosmann, Lars I Kühn

  • 1Microscopy Core Facility, Max Planck Institute for Infection Biology Berlin, Germany.

Frontiers in Immunology
|January 15, 2013
PubMed
Summary
This summary is machine-generated.

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Neutrophil Extracellular Traps (NETs) formation can now be quantified using a novel semi-automatic protocol. This method utilizes anti-chromatin antibodies and DNA-binding dyes for accurate measurement of netting neutrophils.

Area of Science:

  • Immunology
  • Cell Biology

Background:

  • Neutrophil Extracellular Traps (NETs) are formed by neutrophils (polymorphonuclear leukocytes, PMN) through a cell death process called NETosis.
  • NETosis can be triggered by pathogens, platelets, or pathogen components, involving receptor stimulation, ERK pathway activation, and reactive oxygen species (ROS) production.
  • Existing methods for NET quantification, such as fluorescence microscopy and DNA release assays, have limitations including subjectivity and lack of specificity.

Purpose of the Study:

  • To present a protocol for semi-automatic quantification of NET formation.
  • To offer a more objective and reliable method for measuring NETosis compared to existing techniques.

Main Methods:

  • The protocol involves using anti-chromatin antibodies that bind to decondensed chromatin in both the nucleus and NETs.
Keywords:
NEtosischromatinimmunofluorescencequantificationsegmentation

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Real-Time High Throughput Technique to Quantify Neutrophil Extracellular Traps Formation in Human Neutrophils
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Real-Time High Throughput Technique to Quantify Neutrophil Extracellular Traps Formation in Human Neutrophils

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Related Experiment Videos

Last Updated: May 15, 2026

Automated Image-Based Quantification of Neutrophil Extracellular Traps Using NETQUANT
07:33

Automated Image-Based Quantification of Neutrophil Extracellular Traps Using NETQUANT

Published on: November 27, 2019

A High-throughput Assay to Assess and Quantify Neutrophil Extracellular Trap Formation
09:59

A High-throughput Assay to Assess and Quantify Neutrophil Extracellular Trap Formation

Published on: January 29, 2019

Real-Time High Throughput Technique to Quantify Neutrophil Extracellular Traps Formation in Human Neutrophils
07:19

Real-Time High Throughput Technique to Quantify Neutrophil Extracellular Traps Formation in Human Neutrophils

Published on: December 1, 2023

  • Quantification is achieved by relating the fluorescence signal of anti-chromatin antibodies to that of a DNA-binding dye.
  • The method allows for automatic calculation of the percentage of neutrophils undergoing NETosis and requires only standard microscopic equipment and public-domain software.

    • Main Results:

    • The developed protocol enables semi-automatic quantification of NET formation.
    • This method overcomes the subjectivity of fluorescence microscopy and the specificity issues of DNA release assays.
    • The percentage of netting neutrophils can be automatically calculated, providing a more accurate assessment of NETosis.

    Conclusions:

    • A novel, semi-automatic protocol for quantifying NET formation has been established.
    • This method offers improved accuracy and objectivity in assessing NETosis.
    • The protocol is accessible, requiring standard equipment and public-domain software for image analysis.