Nicholas Katherine J, Greenplate Allison R, Flaherty David K, Matlock Brittany K, Juan Juan San, Smith Rita M, Irish Jonathan M, Kalams Spyros A
Department of Pathology, Microbiology and Immunology, Vanderbilt University School of Medicine, Nashville, Tennessee.
Department of Cancer Biology and Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, Tennessee.
Cytometry A. 2016 Mar;89(3):271-80. doi: 10.1002/cyto.a.22799. Epub 2015 Nov 24.
Mass and fluorescence cytometry are quantitative single cell flow cytometry approaches that are powerful tools for characterizing diverse tissues and cellular systems. Here mass cytometry was directly compared with fluorescence cytometry by studying phenotypes of healthy human peripheral blood mononuclear cells (PBMC) in the context of superantigen stimulation. One mass cytometry panel and five fluorescence cytometry panels were used to measure 20 well-established lymphocyte markers of memory and activation. Comparable frequencies of both common and rare cell subpopulations were observed with fluorescence and mass cytometry using biaxial gating. The unsupervised high-dimensional analysis tool viSNE was then used to analyze data sets generated from both mass and fluorescence cytometry. viSNE analysis effectively characterized PBMC using eight features per cell and identified similar frequencies of activated CD4+ T cells with both technologies. These results suggest combinations of unsupervised analysis programs and extended multiparameter cytometry will be indispensable tools for detecting perturbations in protein expression in both health and disease.
质谱流式细胞术和荧光流式细胞术是定量单细胞流式细胞术方法，是表征各种组织和细胞系统的强大工具。在这里，通过在超抗原刺激的背景下研究健康人外周血单个核细胞（PBMC）的表型，将质谱流式细胞术与荧光流式细胞术进行了直接比较。使用一个质谱流式细胞术面板和五个荧光流式细胞术面板来测量20种成熟的记忆和活化淋巴细胞标志物。使用双轴门控，在荧光和质谱流式细胞术中观察到常见和罕见细胞亚群的频率相当。然后使用无监督高维分析工具viSNE来分析由质谱和荧光流式细胞术生成的数据集。viSNE分析使用每个细胞八个特征有效地表征了PBMC，并通过两种技术确定了活化CD4 + T细胞的相似频率。这些结果表明，无监督分析程序和扩展多参数流式细胞术的组合将是检测健康和疾病中蛋白质表达扰动的不可或缺的工具。
