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Validation of CDr15 as a new dye for detecting neutrophil extracellular trap
- Source :
- Biochemical and Biophysical Research Communications. 527:646-653
- Publication Year :
- 2020
- Publisher :
- Elsevier BV, 2020.
-
Abstract
- Neutrophil extracellular trap (NET) is one of the first-line defenses against microbes. Under certain circumstances, however, it also plays an aggravating factor in diverse inflammation-related diseases including cancers and vascular diseases. Our aim is to develop a new method to detect NET in cells and tissues using a DNA-specific fluorescence probe CDr15. CDr15 was characterized to be impermeable to the cell membranes and to emit a strong fluorescence in association with extracellular DNAs in NET. Due to these properties, CDr15 was successfully shown to quantify NETs in vitro and to be applicable for real-time monitoring NET formation in PMA-stimulated neutrophils. Even in formaldehyde-fixed tumor specimens, CDr15 could detect NETs spreading around cancer cells. Compared with DAPI and SYTOX DNA dyes, CDr15 showed a lower level of background fluorescence and a higher specificity in NET detection. Based on these results, we propose CDr15 as a novel marker of NET to be applicable in experimental and clinical studies.
- Subjects :
- 0301 basic medicine
Fluorescence-lifetime imaging microscopy
Neutrophils
Cell
Biophysics
Extracellular Traps
Biochemistry
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
Neoplasms
medicine
Extracellular
Humans
DAPI
Molecular Biology
Cells, Cultured
Fluorescent Dyes
Chemistry
DNA
Cell Biology
Neutrophil extracellular traps
Fluorescence
030104 developmental biology
Membrane
medicine.anatomical_structure
Microscopy, Fluorescence
030220 oncology & carcinogenesis
Cancer cell
Subjects
Details
- ISSN :
- 0006291X
- Volume :
- 527
- Database :
- OpenAIRE
- Journal :
- Biochemical and Biophysical Research Communications
- Accession number :
- edsair.doi.dedup.....4b2b07c47c8996305485b4b8ba4e0ac0