Department of Medicine, Division of Hematology, The Ohio State University Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio, USA.
Cytometry A. 2021 Oct;99(10):1042-1053. doi: 10.1002/cyto.a.24308. Epub 2021 Feb 23.
The identification and discrimination of viable cells is important to understand how experimental variables may influence biochemical processes such as cell metabolism, cell cycle, and signaling pathways. Cisplatin is commonly used as a viability stain in mass cytometry studies, however, recent work by Mei et al. has demonstrated that cisplatin can also be used to label antibodies, complicating the simultaneous use of the platinum measurement channels for both antibody and viability staining. This study demonstrates that other metal salts (hafnium chloride, niobium chloride, and zirconium chloride) can serve as substitutes for cisplatin in viability staining. These stains yield similar fractions of live and dead cells and stain the same dead cells in parallel high parameter analyses. In addition, this study demonstrates how a variety of protein antigen viability markers (pRb, Ki-67, Histone H1, cleaved PARP, and GAPDH) can be used to discriminate live and dead cell populations, without the need for a separate viability staining step. As few as two of these protein antigen viability markers can help identify live and dead cell populations in fixed samples and can identify the same viable cells in high dimensional analyses with or without use of viability stain information. This study demonstrates several alternative approaches to mass cytometry viability assessment that can facilitate use of platinum isotopes for antibody staining and enables identification of live and dead cell populations in samples for which a separate viability stain is not practical.
鉴定和区分活细胞对于了解实验变量如何影响细胞代谢、细胞周期和信号通路等生化过程非常重要。顺铂通常被用作液质联用细胞术研究中的活细胞染色剂，然而，最近 Mei 等人的研究表明，顺铂也可用于标记抗体，这使得同时使用铂测量通道进行抗体和活细胞染色变得复杂。本研究表明，其他金属盐（氯化铪、氯化铌和氯化锆）可替代顺铂用于活细胞染色。这些染色剂产生相似比例的死活细胞，并在平行的高参数分析中对相同的死细胞进行染色。此外，本研究还展示了如何使用多种蛋白质抗原活细胞标志物（pRb、Ki-67、组蛋白 H1、cleaved PARP 和 GAPDH）来区分死活细胞群体，而无需进行单独的活细胞染色步骤。仅使用其中两种蛋白质抗原活细胞标志物，就可以帮助鉴定固定样本中的死活细胞群体，并可以在不使用活细胞染色信息的情况下，在高维分析中识别相同的活细胞。本研究展示了几种替代的液质联用细胞术活细胞评估方法，这可以促进使用铂同位素进行抗体染色，并能够在没有单独活细胞染色剂的情况下，鉴定样本中的死活细胞群体。
