Mass cytometry analysis of immune cells in the brain
Mass cytometry analysis of immune cells in the brain
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DOI：
10.1038/nprot.2017.155
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发表时间：
2018-02-01
期刊：
影响因子：
14.8
通讯作者：
Rolls, Asya
中科院分区：
文献类型：
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作者：
Korin, Ben;Dubovik, Tania;Rolls, Asya
免疫细胞包括一个多样化和动态的细胞群，负责广泛的免疫活动。它们通过细胞因子介导的通讯和直接的细胞间相互作用与其他免疫细胞和非免疫细胞协同作用。理解复杂的免疫网络需要对其单个细胞成分进行广泛的表征。这与脑隔室特别相关，这是一个活跃的免疫部位，由驻留和浸润的免疫细胞组成，影响大脑发育、组织稳态和神经元活动。大规模细胞术或CyTOF（飞行时间细胞术）使用金属偶联抗体在单细胞水平上对数十种标记物进行高维描述，从而提供免疫系统的鸟瞰图。这项技术已经成功地应用于人类和啮齿动物中新的免疫群体的发现。在这里，我们提供了一个逐步描述的质量细胞术方法的分析小鼠脑室。该程序的不同阶段包括脑灌注，脑组织提取并将其解离为单细胞悬浮液，随后用金属标记的抗体对细胞进行染色，使用流式细胞仪读取样本，使用SPADE和viSNE进行数据分析。这个过程需要
Immune cells comprise a diverse and dynamic cell population that is responsible for a broad range of immunological activities. They act in concert with other immune and nonimmune cells via cytokine-mediated communication and direct cell-cell interactions. Understanding the complex immune network requires a broad characterization of its individual cellular components. This is especially relevant for the brain compartment, which is an active immunological site, composed of resident and infiltrating immune cells that affect brain development, tissue homeostasis and neuronal activity. Mass cytometry, or CyTOF (cytometry by time-of-flight), uses metal-conjugated antibodies to enable a high-dimensional description of tens of markers at the single-cell level, thereby providing a bird's-eye view of the immune system. This technique has been successfully applied to the discovery of novel immune populations in humans and rodents. Here, we provide a step-by-step description of a mass cytometry approach for the analysis of the mouse brain compartment. The different stages of the procedure include brain perfusion, extraction of the brain tissue and its dissociation into a single-cell suspension, followed by cell staining with metal-tagged antibodies, sample reading using a mass cytometer, and data analysis using SPADE and viSNE. This procedure takes