Axe des Maladies Infectieuses et Immunitaires, Centre de Recherche du CHU de Québec-Université Laval, Pavillon CHUL, Québec (QC), G1V 4G2, Canada.
Methods Appl Fluoresc. 2018 May 29;6(3):037002. doi: 10.1088/2050-6120/aac51b.
Extracellular traps ejected by various immune cells (neutrophils, macrophages, eosinophils and mast cells) have several immune functions, either protective against pathogens or deleterious in some autoimmune or inflammatory disorders. Since their first description in 2004, the mechanisms of extracellular traps formation have been extensively investigated though still not fully understood. We describe here a new tool for the detection of extracellular traps by fluorescence microscopy in a single-step staining protocol, which does not require any wash. The approach uses the GreenGlo™ DNA dye, which can differentiate between nuclear DNA and extracellular DNA (extracellular traps) released from cells using different fluorescence excitation wavelengths. GreenGlo™ staining is suitable for adherent and non-adherent cells and is expected to be extendable to extracellular traps from other cells types (i.e. eosinophils, mast cells and monocytes).
各种免疫细胞（中性粒细胞、巨噬细胞、嗜酸性粒细胞和肥大细胞）释放的细胞外陷阱具有多种免疫功能，无论是针对病原体的保护作用，还是在某些自身免疫或炎症性疾病中的有害作用。自 2004 年首次描述以来，尽管对细胞外陷阱形成的机制进行了广泛的研究，但仍未完全了解。我们在这里描述了一种新的荧光显微镜检测细胞外陷阱的工具，该工具在一个单步染色方案中使用，不需要任何洗涤。该方法使用 GreenGlo™ DNA 染料，该染料可以使用不同的荧光激发波长区分细胞核 DNA 和从细胞释放的细胞外 DNA（细胞外陷阱）。GreenGlo™ 染色适用于贴壁和非贴壁细胞，并有望扩展到其他细胞类型（即嗜酸性粒细胞、肥大细胞和单核细胞）的细胞外陷阱。
