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对源自小鼠骨髓的中性粒细胞的中性粒细胞胞外诱捕网(NETs)进行定量和可视化分析。

Quantification and visualization of neutrophil extracellular traps (NETs) from murine bone marrow-derived neutrophils.

作者信息

Vong Linda, Sherman Philip M, Glogauer Michael

机构信息

Faculty of Medicine, University of Toronto, Toronto, ON, Canada.

出版信息

Methods Mol Biol. 2013;1031:41-50. doi: 10.1007/978-1-62703-481-4_5.

Abstract

Neutrophils are one of the first cells to respond to an inflammatory stimulus, and are equipped with an assortment of antimicrobial and proteolytic enzymes to disarm and degrade bacterial pathogens. A novel mechanism of bacterial trapping, termed neutrophil extracellular traps (NETs), was recently described whereby neutrophils were shown to cast out web-like structures of chromatin, capturing and immobilizing invading pathogens. Herein we describe protocols to isolate murine bone marrow-derived neutrophils, and spectrophotometrically quantify, immunolabel, and visualize NET structures in vitro.

摘要

中性粒细胞是最早对炎症刺激作出反应的细胞之一,它们配备了各种抗菌和蛋白水解酶,以解除细菌病原体的武装并将其降解。最近描述了一种新的细菌捕获机制,称为中性粒细胞胞外陷阱(NETs),据此显示中性粒细胞会抛出染色质的网状结构,捕获并固定入侵的病原体。在此,我们描述了分离小鼠骨髓来源中性粒细胞的方法,以及通过分光光度法定量、免疫标记和体外观察NET结构的方法。

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