Flow cytometry and fluorescence-activated cell sorting have become invaluable tools to analyze and isolate specific cell populations in a wide range of biomedical research and clinical applications. In countless approaches worldwide, scientists are using single cell analyses to better understand the significance and variation within different cellular populations, and fluorescence-activated cell sorting has become a major technique for cell isolation in both basic and clinical research. However, majority of available cell sorters are pressurized, droplet-based systems, which apply significant environmental pressure and shear stress to cells during sorting. Recently, the flow cytometry community has become increasingly aware about the potential negative effects this could have on sorted cells and the term “sorter induced cell stress” (SICS) has been proposed. However, up to date only a limited number of studies have investigated the effects of cell sorting on cell viability and function. Therefore, solid data on the effects of sheath pressure and nozzle size on survival and function of sorted cells are surprisingly rare. With this in mind, we sorted “CD4⁺” T-cells and “live” cells from human peripheral blood mononuclear cells (PBMCs) at different sort conditions and analyzed their quality before and after sorting in a series of assays. Here we present our findings in reference to cell viability and cell proliferation following sorting on different instruments (BD FACSAria III SORP and BD FACSJazz), utilizing different nozzle sizes (70 to 100 μm) and sheath pressure settings (20 to 70 psi). The results show no significant differences in cell viability and proliferation after the different tested sort conditions, but rather differences between individual experiments. These findings are evaluated and their potential significance in cell sorting experiments is discussed.

流式细胞术和荧光激活的细胞分选已成为在广泛的生物医学研究和临床应用中分析和分离特定细胞群的宝贵工具。在世界范围内，无数方法中，科学家正在使用单细胞分析来更好地了解不同细胞群体内的意义和变异，并且荧光激活的细胞分选已成为基础和临床研究中用于细胞分离的主要技术。但是，大多数可用的细胞分选仪都是基于液滴的加压系统，该系统在分选过程中会对细胞施加很大的环境压力和剪切应力。最近，流式细胞仪界越来越意识到这可能对分选的细胞产生潜在的负面影响，因此提出了术语“分选诱导的细胞应激”（SICS）。然而，迄今为止，仅有限数量的研究调查了细胞分选对细胞活力和功能的影响。因此，关于鞘管压力和喷嘴尺寸对分选细胞的存活和功能影响的可靠数据非常少见。考虑到这一点，我们对“ CD4⁺来自人类外周血单核细胞（PBMC）在不同分选条件下的T细胞和“活”细胞，并在一系列分析中对分选前后的质量进行了分析。在这里，我们介绍了在不同仪器（BD FACSAria III SORP和BD FACSJazz）上进行分类后，利用不同的喷嘴尺寸（70至100μm）和鞘管压力设置（20至70 psi）对细胞活力和细胞增殖的发现。结果表明，在不同的测试分类条件下，细胞活力和增殖没有显着差异，但各个实验之间却存在差异。对这些发现进行了评估，并讨论了它们在细胞分选实验中的潜在意义。