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Updated: Jan 16, 2026
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Automated Image-Based Quantification of Neutrophil Extracellular Traps Using NETQUANT
Published on: November 27, 2019
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Tissue Cytometry Assay with Nuclear Segmentation for Quantifying NETotic Cells in Neutrophils Stimulated by
Rodrigo Rivera-Concha1,2, Marion León3, Nikol Ponce-Rojas4,5
1Center of Excellence in Translational Medicine-Scientific and Technological Bioresource Nucleus (CEMT-BIOREN), Edificio Biociencias de la Salud, Faculty of Medicine, Universidad de La Frontera, Avenida Alemania 0458, Temuco 4810296, Chile.
Animals : an Open Access Journal From MDPI
|September 27, 2025
Summary
An automated method quantifies neutrophil nuclear area expansion, a key indicator of neutrophil activation. This technique reliably measures neutrophil extracellular trap release in canine and bovine species.
Area of Science:
- Immunology
- Cell Biology
- Veterinary Science
Background:
- Neutrophils are critical immune cells with functions including phagocytosis, degranulation, and neutrophil extracellular trap (NET) release.
- Measuring neutrophil nuclear area expansion is a standard method for assessing early neutrophil activation.
- Existing methods for quantifying nuclear area expansion can be time-consuming and require manual analysis.
Purpose of the Study:
- To develop and validate an automated method for measuring neutrophil nuclear area expansion in canine and bovine species.
- To assess the utility of this automated method for detecting neutrophil extracellular trap release.
Main Methods:
- Neutrophils were isolated from canine and bovine blood.
- Neutrophils were co-incubated with spermatozoa (fresh for canine, cryopreserved for bovine).
- Fluorescence microscopy (TissueFAXS) and automated image analysis software (StrataQuest v.7.0) were used to measure nuclear area expansion and co-localization of neutrophil elastase and DNA.
Main Results:
- The automated method successfully detected neutrophil nuclear area expansion in both canine and bovine species upon activation with spermatozoa.
- Nuclear expansion was observed as early as 15 minutes and persisted up to 120 minutes.
- Analysis confirmed reliable and consistent data generation, comparable to previously published methods.
Conclusions:
- An automated method for measuring neutrophil nuclear area expansion has been successfully developed and validated for canine and bovine species.
- This automated approach provides a reliable and efficient alternative for assessing neutrophil activation and NETosis.
- The method facilitates the study of neutrophil function across different mammalian species.
