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A High-throughput Assay to Assess and Quantify Neutrophil Extracellular Trap Formation
JoVE Journal
Immunologia e infezioni
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JoVE Journal Immunologia e infezioni
A High-throughput Assay to Assess and Quantify Neutrophil Extracellular Trap Formation

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

DOI:
10.3791/59150-v

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09:59 min

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January 29, 2019

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, , , , , ,
1Department of Nephrology,Leiden University Medical Center

Capitoli

  • 00:04Titolo
  • 01:13Isolation of Healthy Neutrophils
  • 03:42Red Fluorescent Cell Labelling of Neutrophils
  • 05:10Induction of Neutrophil Extracellular Trap Formation
  • 06:42NET Visualization with 3D High-content, High-resolution Immunofluorescence Confocal Microscopy and Analysis of NET Formation
  • 07:59Results: Assessment and Quantification of Neutrophil Extracellular Trap Formation
  • 09:26Conclusion

Summary

Traduzione automatica

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This protocol describes a highly sensitive and high throughput neutrophil extracellular trap (NET) assay for the semi-automated quantification of ex vivo NET formation by immunofluorescence three-dimensional confocal microscopy. This protocol can be used to evaluate NET formation and degradation after different stimuli and can be used to study potential NET-targeted therapies.

Tags

Keywords: Neutrophil Extracellular Traps (NETs)High-throughput AssayImmunofluorescence Confocal MicroscopyAutoimmune DiseasesANCA-associated VasculitisSystemic Lupus ErythematosusNeutrophil IsolationDensity Gradient Centrifugation

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