Flow Cytometry Analysis to Identify Human CD4+ T Cell Subsets
Flow Cytometry Analysis to Identify Human CD4+ T Cell Subsets
复制标题
DOI：
10.1007/978-1-4939-9728-2_2
复制
发表时间：
2019-01-01
期刊：
影响因子：
--
通讯作者：
Gorry, Paul
中科院分区：
文献类型：
--
作者：
Flynn, Jacqueline;Gorry, Paul
流式细胞术是一种强大的工具，它使用激光来分析细胞的各种不同特征。它通常用于确定细胞表面标志物和细胞内分子的表达，以使用细胞大小、粒度和荧光标记抗体将细胞定义为不同的群体。因此，流式细胞术能够同时对单细胞进行多参数分析。在染色过程中，创建单细胞悬浮液，用流式细胞术抗体进行染色，以便在流式细胞仪上进行分析。通过改变流式抗体靶向的表面标记，该技术的染色方法可用于不同的细胞类型，前提是所有抗体在使用前都经过滴定，并在了解用于检测的表面分子的密度和荧光染料的亮度的情况下进行选择，以指导抗体选择。本章的方案专门用于检测人 CD4(+) T 细胞亚群，通过表面标记表型分析定义幼稚和记忆亚群。
Flow cytometry is a powerful tool, which uses lasers to analyze a wide range of different characteristics of cells. It is commonly used to determine the expression of cell surface markers and intracellular molecules to define cells into different populations using cell size, granularity, and fluorescently labeled antibodies. Thus, flow cytometry enables simultaneous and mutliparameter analysis of single cells.During the staining procedure, a single cell suspension is created for staining with flow cytometry antibodies for analysis on the flow cytometer. The staining methods from this technique can be used for different cell types by changing the surface marker targeted by the flow antibody, provided all antibodies are titrated prior to use, and are chosen with knowledge of the density of surface molecule for detection and brightness of fluorochrome to guide antibody selection.This chapter's protocol has been designed specifically for detection of human CD4(+) T cell subsets defining naive and memory subpopulations by surface marker phenotyping.