应用液质联用技术分析人类全血中的细胞表型和信号状态
Mass Cytometry Assessment of Cell Phenotypes and Signaling States in Human Whole Blood.
机构信息
Institute for Immunity, Transplantation, and Infection, Stanford University School of Medicine, Stanford, CA, USA.
出版信息
Methods Mol Biol. 2022;2543:113-128. doi: 10.1007/978-1-0716-2553-8_10.
Phosphoflow is a powerful tool that allows researchers to measure distinct signaling responses to various stimuli in multiple subpopulations of cells. Extension of this technique to mass cytometry (cytometry by time-of-flight or CyTOF) allows many more cell phenotypes and signaling nodes to be interrogated in parallel. The use of fresh whole blood is ideal for capturing the in vivo signaling state of all leukocytes, including granulocytes. In this chapter, we provide a detailed protocol for performing CyTOF phosphoflow in human whole blood, using cytokines and other stimuli. Barcoding and combining of multiple samples and other techniques to reduce batch effects and provide optimal comparability between samples/stimulations are also described.
磷酸化流式技术是一种强大的工具，它允许研究人员测量多种细胞亚群对不同刺激的不同信号反应。将该技术扩展到液质流式细胞术（飞行时间或 CyTOF 细胞术）中，可以同时检测更多的细胞表型和信号节点。使用新鲜的全血是捕获所有白细胞（包括粒细胞）体内信号状态的理想选择。在本章中，我们提供了一个详细的协议，用于在人类全血中进行 CyTOF 磷酸化流式技术，使用细胞因子和其他刺激物。我们还描述了对多个样本进行条形码标记和组合，以及其他技术，以减少批次效应并提供样本/刺激之间的最佳可比性。
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