A novel image-based quantitative method for the characterization of NETosis
Wenpu Zhao1, Darin K Fogg2, Mariana J Kaplan1
1Systemic Autoimmunity Branch, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, MD 20892, United States.
Journal of Immunological Methods
|May 25, 2015
Summary
Researchers developed a novel method to quantify both
Area of Science:
- Immunology
- Cell Biology
Background:
- Neutrophil extracellular trap (NET) formation, or NETosis, is a programmed cell death mechanism involving DNA release.
- Two forms, 'suicidal' and 'vital' NETosis, have been described, but methods for their simultaneous assessment are lacking.
- This hinders detailed characterization and biomarker development for NETosis.
Purpose of the Study:
- To develop a novel, rapid, quantitative, and semiautomated method for simultaneously assessing both 'suicidal' and 'vital' NETosis.
- To enable detailed characterization of NETosis mechanisms and explore its potential as a biomarker.
Main Methods:
- Utilized high-speed multi-spectral imaging coupled with morphometric image analysis.
- Employed imaging flow cytometry for automated analysis of subcellular morphology and texture.
- Quantified spontaneous and stimulus-induced NET formation ex-vivo and in vitro.
Main Results:
- Successfully developed a method to simultaneously assess 'suicidal' and 'vital' NETosis.
- The method provides automated, quantitative, and rapid analysis of NET formation.
- Demonstrated the potential for NETosis assessment as a biomarker in pre-clinical and clinical studies.
Conclusions:
- The new imaging flow cytometry-based method enables simultaneous, quantitative assessment of both 'suicidal' and 'vital' NETosis.
- This advancement facilitates deeper understanding of NETosis mechanisms.
- The method holds promise for NETosis biomarker development in various disease contexts.
