使用质谱流式细胞术进行高参数免疫分析
High-Parameter Immune Profiling with CyTOF.
作者信息
Sahaf Bita, Rahman Adeeb, Maecker Holden T, Bendall Sean C
机构信息
PICI Cancer Correlative Science Unit, Cancer Institute, Stanford University School of Medicine, Stanford, CA, USA.
Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, USA.
出版信息
Methods Mol Biol. 2020;2055:351-368. doi: 10.1007/978-1-4939-9773-2_16.
Mass cytometry, or CyTOF, is a useful technology for high-parameter single-cell phenotyping, especially from suspension cells such as blood or PBMC. It is particularly appealing to monitor the systemic immune changes that could accompany cancer immunotherapy. Here we present a reference panel for identification of all major immune cell populations, with flexibility for addition of trial-specific markers. We also describe best-practice measures for minimizing and tracking batch variability. These include: sample barcoding, use of spiked-in reference cells, and lyophilization of the antibody cocktail. Our protocol assumes the use of cryopreserved PBMC, both for convenience of batching samples and for maximum comparability across patients and time points. Finally, we show an option for automated analysis using the Astrolabe platform (Astrolabe Diagnostics, Inc.).
质谱流式细胞术（Mass cytometry），即细胞索引质谱流式技术（CyTOF），是一种用于高参数单细胞表型分析的实用技术，尤其适用于对血液或外周血单核细胞（PBMC）等悬浮细胞进行分析。它对于监测癌症免疫治疗过程中可能伴随的全身免疫变化特别有吸引力。在此，我们展示了一个用于识别所有主要免疫细胞群体的参考面板，并可灵活添加特定试验的标志物。我们还描述了将批次变异性降至最低并进行跟踪的最佳实践措施。这些措施包括：样本条形码标记、使用掺入的参考细胞以及抗体鸡尾酒的冻干。我们的方案假定使用冷冻保存的PBMC，这既便于批量处理样本，又能在患者和时间点之间实现最大程度的可比性。最后，我们展示了一种使用Astrolabe平台（Astrolabe诊断公司）进行自动化分析的方法。
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