Cell viability is a physiological status connected to cell membrane integrity and cytoplasmic topography, which is profoundly important for fundamental biological research and practical biomedical applications. A conventional method for assessing cell viability is through cell staining analysis. However, cell staining involves laborious and complicated processing procedures and is normally cytotoxic. Intrinsic cellular phenotypes thus provide new avenues for measuring cell viability in a stain-free and non-toxic manner. In this work, we present a label-free non-destructive impedance-based approach for cell viability assessment by simultaneously characterizing multiple electrical cellular phenotypes in a high-throughput manner (>1000 cells per min). A novel concept called the complex opacity spectrum is introduced for improving the discrimination of live and dead cells. The analysis of the complex opacity spectrum leads to the discovery of two frequency ranges that are optimized for characterizing membranous and cytoplasmic electrical phenotypes. The present impedance-based approach has successfully discriminated between living and dead cells in two different experimental scenarios, including mixed living and dead cells in both homogenous and heterogeneous cell samples. This impedance-based single cell phenotyping technique provides highly accurate and consistent cell viability analysis, which has been validated by commercial fluorescence-based flow cytometry (∼1% difference) using heterogeneous cell samples. This label-free high-throughput cell viability analysis strategy will have broad applications in the field of biology and medicine.

中文翻译：

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多频单细胞电阻抗测量，用于无标记细胞生存力分析

细胞活力是一种与细胞膜完整性和胞质形貌有关的生理状态，这对于基础生物学研究和实际生物医学应用而言极为重要。评估细胞活力的常规方法是通过细胞染色分析。然而，细胞染色涉及费力且复杂的处理过程，并且通常具有细胞毒性。因此，固有的细胞表型提供了以无污染且无毒的方式测量细胞活力的新途径。在这项工作中，我们通过以高通量方式（每分钟> 1000个细胞）同时表征多种细胞电表型，提出了一种基于无标签，基于非破坏性阻抗的细胞活力评估方法。为了改善对活细胞和死细胞的分辨力，引入了一种称为复杂不透明光谱的新概念。复杂的不透明度频谱的分析导致发现两个频率范围的发现，这两个频率范围已被优化以表征膜和细胞质的电表型。本基于阻抗的方法已成功区分了两种不同实验场景下的活细胞和死细胞，包括均质和异质细胞样品中的活细胞和死细胞混合。这种基于阻抗的单细胞表型分析技术可提供高度准确且一致的细胞活力分析，已通过使用异质细胞样品的商用荧光流式细胞仪（相差约1％）进行了验证。