Flow cytometrists have long observed a spectrum of cell-type-specific changes ranging from minor functional defects to outright cell destruction after purification of cells using conventional droplet cell sorters. We have described this spectrum of cell perturbations as sorter induced cellular stress, or SICS (Lopez and Hulspas, Cytometry, 2020, 97, 105–106). Despite the potential impact of this issue and ubiquitous anecdotes, little has been reported about this phenomenon in the literature, and the underlying mechanism has been elusive. Inspired by others' observations (Llufrio et al., Redox Biology, 2018, 16, 381–387 and Binek et al., Journal of Proteome Research, 2019, 18, 169–181), we set out to examine SICS at the metabolic level and use this information to propose a working model. Using representative suspension (Jurkat) and adherent (NIH/3T3) cell lines we observed broad and consistent metabolic perturbations after sorting using a high-speed droplet cell sorter. Our results suggest that the SICS metabolic phenotype is a common cell-type-independent manifestation and may be the harbinger of a wide-range of functional defects either directly related to metabolism, or cell stress response pathways. We further demonstrate a proof of concept that a modification to the fluidic environment (complete media used as sheath fluid) in a droplet cell sorter can largely rescue the intracellular markers of SICS, and that this rescue is not due to a contribution of metabolites found in media. Future studies will focus on characterizing the potential electro-physical mechanisms inherent to the droplet cell sorting process to determine the major contributors to the SICS mechanism.

中文翻译：

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鞘液会影响受分选诱导细胞应激 (SICS) 影响的细胞中细胞代谢物的消耗

流式细胞仪长期以来一直观察到一系列细胞类型特异性变化，从轻微的功能缺陷到使用传统液滴细胞分选仪纯化细胞后的彻底细胞破坏。我们已将这种细胞扰动谱描述为分拣机诱导的细胞应激或 SICS（Lopez 和 Hulspas，Cytometry，2020, 97, 105–106）。尽管这个问题的潜在影响和无处不在的轶事，但文献中关于这种现象的报道很少，而且潜在的机制也难以捉摸。受其他人观察的启发（Llufrio 等人，Redox Biology，2018,16,381-387 和 Binek 等人，蛋白质组研究杂志，2019,18,169-181），我们开始在代谢方面检查 SICS水平并使用此信息提出工作模型。使用具有代表性的悬浮液 (Jurkat) 和贴壁 (NIH/3T3) 细胞系，我们在使用高速液滴细胞分选器分选后观察到广泛且一致的代谢扰动。我们的研究结果表明，SICS 代谢表型是一种常见的与细胞类型无关的表现，可能是与代谢或细胞应激反应途径直接相关的广泛功能缺陷的预兆。我们进一步证明了一个概念证明，即对液滴细胞分选器中的流体环境（用作鞘液的完整介质）的修改可以在很大程度上拯救 SICS 的细胞内标记物，并且这种拯救不是由于在媒体。