细胞的活力，是反映其功能和生理完整性的关键指标。活细胞和死细胞在代谢方面存在着巨大的差异。然而，在以往大多数基于质谱的代谢研究中，这个重要因素常常被忽略，这可能会让我们对生物系统产生误解。为了解决这个问题，科研人员开发出了一种名为“活力信息单细胞质谱技术（ViSCMS）”的新方法。这种技术具有高通量、无需标记的特点，能够同时测量单个细胞的活力和代谢特征。在具体操作中，研究人员利用磷酸胆碱（PC）34:1的信号来确认细胞事件，还找到了谷胱甘肽（GSH）作为区分活细胞和死细胞的内在标记物。通过ViSCMS得到的细胞活力率与传统的吖啶橙/碘化丙啶（AO/PI）染色法高度一致，平均偏差仅为0.33%，而且在不同活力率的多种细胞系中都展现出了出色的重现性（标准差小于3%）。这项技术的实用性也得到了充分验证。研究人员成功地对六种结直肠癌细胞系进行了亚型分类，而这只有在考虑细胞活力的情况下才能实现。此外，在研究抗癌药物对HCT116细胞的作用时，还观察到了明显的与细胞活力相关的代谢差异，这为我们从单细胞水平了解肿瘤细胞如何适应化疗压力提供了新的视角。相信随着这项技术的不断发展和应用，我们将能更全面、深入地揭示细胞代谢的奥秘。

期刊

Analytical Chemistry

标题

Viability-Informed Single-Cell Mass Spectrometry for More Comprehensive Metabolic Analysis

作者

Simin Cheng, Danni Wu, Xinxin Wang, Siyuan Tan, Lulu Feng, Xiaoping Yu, Xiaoyun Gong, Xinhua Dai

摘要

The viability of cells is a key indicator of cellular function and physiological integrity. There are great metabolic differences between live and dead cells. However, most mass spectrometric metabolic studies overlook this issue, leading to a potential misunderstanding of biological systems. Here, we propose the development of a high-throughput and label-free viability-informed single-cell mass spectrometry (ViSCMS) technique for the simultaneous measurement of cell viability and metabolic profiles. The signal of phosphocholine (PC) 34:1 was used for the confirmation of cell events. Glutathione (GSH) was identified as an intrinsic marker to further accurately distinguish live and dead cells. The viability rates obtained by ViSCMS were in good concordance with those of conventional AO/PI staining (mean bias: 0.33%). Excellent reproducibility (SD < 3%) was achieved across multiple cell lines with varied viability rates. The practicality of the method was demonstrated by the successful subtyping of six colorectal cancer (CRC) cell lines. The subtyping could only be achieved when the viability of cells was taken into consideration. Furthermore, clear viability-dependent metabolic differences were observed on HCT116 cells when treated with an anticancer drug. The results provided insights into how tumor cells adapt to chemotherapy stress at the single-cell level.

原文链接

https://pubs.acs.org/doi/10.1021/acs.analchem.5c07083

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