Flow Cytometry Analysis to Identify Human CD4+ T Cell Subsets

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Abstract

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 naïve and memory subpopulations by surface marker phenotyping.

Original languageEnglish
Title of host publicationIn Vitro Differentiation of T-Cells
Subtitle of host publicationMethods and Protocols
EditorsShin Kaneko
Place of PublicationNew York, NY
PublisherHumana Press
Chapter2
Pages15-25
Number of pages11
ISBN (Electronic)9781493997282
ISBN (Print)9781493997275
DOIs
Publication statusPublished - 1 Jan 2019

Publication series

NameMethods in Molecular Biology
Volume2048
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • CD markers
  • CD4 T cells
  • Flow cytometry
  • Memory T cells
  • Naïve T cells
  • T cell subsets

Cite this

Flynn, J., & Gorry, P. (2019). Flow Cytometry Analysis to Identify Human CD4+ T Cell Subsets. In S. Kaneko (Ed.), In Vitro Differentiation of T-Cells: Methods and Protocols (pp. 15-25). (Methods in Molecular Biology; Vol. 2048). Humana Press. https://doi.org/10.1007/978-1-4939-9728-2_2