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Related Experiment Video

Updated: Feb 7, 2026

Immunofluorescence Labelling of Human and Murine Neutrophil Extracellular Traps in Paraffin-Embedded Tissue
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NETs detection and quantification in paraffin embedded samples using confocal microscopy.

A Santos1, P Martín2, A Blasco3

  • 1Molecular Pathology Laboratory, Instituto de Investigación Puerta de Hierro Segovia Arana (IDIPHISA), C/ Joaquín Rodrigo, 2, 28222 Majadahonda, Madrid, Spain.

Micron (Oxford, England : 1993)
|July 21, 2018
PubMed
Summary
This summary is machine-generated.

This study presents a new semi-automatic method for detecting Neutrophil Extracellular Traps (NETs) in thrombus samples using MPO and H3Cit markers. This observer-independent technique aids in standardizing NETs research in various diseases.

Keywords:
Image analysisInmunofluorescenceMicroscopyNeutrophil Extracellular Traps

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Area of Science:

  • Immunology
  • Pathology
  • Cardiovascular Research

Background:

  • Neutrophil Extracellular Traps (NETs) are increasingly recognized for their pathological roles in diverse diseases.
  • Accurate detection and quantification of NETs in tissue samples are crucial for understanding disease mechanisms.
  • Existing methods for NETs analysis in tissues can be subjective and lack standardization.

Purpose of the Study:

  • To develop and validate a semi-automatic method for visualizing and quantifying NETs.
  • To apply this method to paraffin-embedded intracoronary thrombus aspirate samples.
  • To establish an observer-independent approach for NETs quantification in tissue research.

Main Methods:

  • Utilized confocal immunofluorescence microscopy for NETs detection.
  • Employed colocalization of myeloperoxidase (MPO) and citrullinated histone 3 (H3Cit) as NETs hallmarks.
  • Quantified NETs by counting positive fields and measuring total NETs area (μm²) in thrombus samples.

Main Results:

  • Successfully visualized and quantified NETs in paraffin-embedded thrombus aspirates.
  • The method demonstrated observer-independent quantification, reducing subjectivity.
  • Established a reproducible approach for NETs analysis in tissue samples.

Conclusions:

  • The described semi-automatic method provides a standardized tool for NETs detection and quantification.
  • This technique is valuable for studying the role of NETs in cardiovascular diseases and other pathologies.
  • Facilitates consistent comparison of NETs findings across different research laboratories.