High-dimensional mass cytometry data potentially enable a comprehensive characterization of immune cells. In order to positively affect clinical trials and translational clinical research, this advanced technology needs to demonstrate a high reproducibility of results across multiple sites for both peripheral blood mononuclear cells (PBMC) and whole blood preparations. A dry 30-marker broad immunophenotyping panel and customized automated analysis software were recently engineered and are commercially available as the Fluidigm® Maxpar® Direct™ Immune Profiling Assay™. In this study, seven sites received whole blood and six sites received PBMC samples from single donors over a 2-week interval. Each site labeled replicate samples and acquired data on Helios™ instruments using an assay-specific acquisition template. All acquired sample files were then automatically analyzed by Maxpar Pathsetter™ software. A cleanup step eliminated debris, dead cells, aggregates, and normalization beads. The second step automatically enumerated 37 immune cell populations and performed label intensity assessments on all 30 markers. The inter-site reproducibility of the 37 quantified cell populations had consistent population frequencies, with an average %CV of 14.4% for whole blood and 17.7% for PBMC. The dry reagent coupled with automated data analysis is not only convenient but also provides a high degree of reproducibility within and among multiple test sites resulting in a comprehensive yet practical solution for deep immune phenotyping.

中文翻译：

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使用 CyTOF 技术结合 Maxpar Pathsetter（一种自动数据分析系统），在全血和外周血单核细胞制剂中进行人体免疫表型分析的多位点重现性。

高维质谱数据可能有助于对免疫细胞进行全面表征。为了对临床试验和转化临床研究产生积极影响，这项先进技术需要证明外周血单核细胞 (PBMC) 和全血制剂在多个位点的结果具有高度可重复性。最近设计了一个干燥的 30 标记广泛免疫表型面板和定制的自动分析软件，并作为 Fluidigm® Maxpar® Direct™ Immune Profiling Assay™ 上市销售。在这项研究中，在 2 周的时间间隔内，七个地点接受了全血，六个地点接受了来自单个供体的 PBMC 样本。每个站点都使用特定于分析的采集模板在 Helios™ 仪器上标记重复样本和采集数据。然后通过 Maxpar Pathsetter™ 软件自动分析所有采集的样品文件。净化步骤消除了碎片、死细胞、聚集体和归一化珠。第二步自动枚举 37 个免疫细胞群，并对所有 30 个标记进行标记强度评估。37 个量化细胞群的位点间重现性具有一致的群体频率，全血的平均 %CV 为 14.4%，PBMC 的平均 %CV 为 17.7%。干试剂与自动数据分析相结合，不仅方便，而且在多个测试点内部和之间提供高度的可重复性，从而为深度免疫表型分析提供全面而实用的解决方案。第二步自动枚举 37 个免疫细胞群，并对所有 30 个标记进行标记强度评估。37 个量化细胞群的位点间重现性具有一致的群体频率，全血的平均 %CV 为 14.4%，PBMC 的平均 %CV 为 17.7%。干试剂与自动数据分析相结合，不仅方便，而且在多个测试点内部和之间提供高度的可重复性，从而为深度免疫表型分析提供全面而实用的解决方案。第二步自动枚举 37 个免疫细胞群，并对所有 30 个标记进行标记强度评估。37 个量化细胞群的位点间重现性具有一致的群体频率，全血的平均 %CV 为 14.4%，PBMC 的平均 %CV 为 17.7%。干试剂与自动数据分析相结合，不仅方便，而且在多个测试点内部和之间提供高度的可重复性，从而为深度免疫表型分析提供全面而实用的解决方案。