巨噬细胞和凋亡细胞(AC)的体外共培养为研究有效的细胞增殖提供了实用且有用的工具。这里,我们描述了一种使用成像流式细胞术(IFC)对原代巨噬细胞识别和吞噬凋亡细胞进行自动定量和成像的方法。基于IFC的分析使我们能够成功地量化红细胞增多,明确区分吞噬和非吞噬巨噬细胞,更重要的是,从那些处于认可阶段的人来看,这是标准流式细胞分析无法实现的。为此,我们建立了一个普遍适用的分析管道,以解决可以很容易地适应任何巨噬细胞群体从不同来源的样本。
In vitro cocultures of macrophages and apoptotic cells (ACs) provide a practical and useful tool to study efferocytosis. Here, we describe a method for automated quantification and imaging of recognition and engulfment of apoptotic cells by primary macrophages using imaging flow cytometry (IFC). IFC-based analysis allows us to successfully quantify efferocytosis, clearly distinguishing phagocytic from nonphagocytic macrophages and, more importantly, from those in recognition stage, which is not achievable by standard flow cytometrical analysis. To this end, we established a universally employable analysis pipeline to address efferocytosis that can be easily adapted to any macrophage population from samples of different origins.