Department of Infectious Diseases and Hospital Epidemiology, University Hospital Zurich and University of Zurich, Zurich, Switzerland.
Methods Mol Biol. 2023;2674:235-250. doi: 10.1007/978-1-0716-3243-7_16.
Regulated cell death (RCD) and the concomitant release of extracellular traps by neutrophils (NETs) constitute an important antibacterial effector response. Usually, the dynamic processes of RCD and NETs release are assessed independently of each other by either unspecific or time-consuming methods. Here, we describe a flow cytometry-based high-throughput analysis method incorporating neutrophil RCD and NETs release with visual live-imaging conformation upon ex vivo bacterial challenge. This combined approach allows to quantify and closely follow the kinetics of the dynamic neutrophil effector response towards bacterial infection.
受调控的细胞死亡(RCD)和中性粒细胞(NETs)释放细胞外陷阱是一种重要的抗细菌效应反应。通常,通过非特异性或耗时的方法分别评估 RCD 和 NETs 释放的动态过程。在这里,我们描述了一种基于流式细胞术的高通量分析方法,该方法结合了体外细菌刺激下的中性粒细胞 RCD 和 NETs 释放以及可视化的实时成像确认。这种联合方法可定量和密切跟踪动态中性粒细胞效应反应对细菌感染的动力学。