The characterization of specialized cell subpopulations in a heterogeneous tissue is essential for understanding organ function in health and disease. A popular method of cell isolation is fluorescence-activated cell sorting (FACS) based on probes that bind surface or intracellular markers. In this study, we analyze the impact of FACS on the cell metabolome of mouse peritoneal macrophages. Compared with directly pelleted macrophages, FACS-treated cells had an altered content of metabolites related to the plasma membrane, activating a mechanosensory signaling cascade causing inflammation-like stress. The procedure also triggered alterations related to energy consumption and cell damage. The observed changes mostly derive from the physical impact on cells during their passage through the instrument. These findings provide evidence of FACS-induced biochemical changes, which should be taken into account in the design of robust metabolic assays of cells separated by flow cytometry.

中文翻译：

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流式细胞仪对细胞代谢组有重要影响。

异质组织中特化细胞亚群的表征对于了解健康和疾病中的器官功能至关重要。一种流行的细胞分离方法是基于结合表面或细胞内标记物的探针的荧光激活细胞分选（FACS）。在这项研究中，我们分析了流式细胞仪对小鼠腹膜巨噬细胞细胞代谢组的影响。与直接沉淀的巨噬细胞相比，经FACS处理的细胞的质膜相关代谢产物含量发生了变化，从而激活了机械感觉信号级联反应，从而引起了类似炎症的应激反应。该程序还引发了与能量消耗和细胞损伤有关的改变。观察到的变化主要来自细胞通过仪器时对细胞的物理影响。