Deep Immunophenotyping of Human Whole Blood by Standardized Multi-parametric Flow Cytometry Analyses

被引:9
|
作者
Gao, Jian [1 ,2 ,3 ]
Luo, Yali [1 ,2 ,3 ]
Li, Helian [1 ,2 ,3 ]
Zhao, Yiran [1 ,2 ,3 ]
Zhao, Jialin [1 ,2 ,3 ]
Han, Xuling [1 ,2 ,3 ]
Han, Jingxuan [1 ,2 ,3 ]
Lin, Huiqin [1 ]
Qian, Feng [1 ,2 ,3 ,4 ]
机构
[1] Fudan Univ, Human Phenome Inst, Shanghai Publ Hlth Clin Ctr, State Key Lab Genet Engn,Zhangjiang Fudan Int Inno, Shanghai 200438, Peoples R China
[2] Fudan Univ, Sch Life Sci, Shanghai 200438, Peoples R China
[3] Fudan Univ, Sch Life Sci, Minist Educ, Key Lab Contemporary Anthropol, Shanghai 200438, Peoples R China
[4] Int Human Phenome Inst Shanghai, Inst Immunophenome, Shanghai 200433, Peoples R China
来源
PHENOMICS | 2023年 / 3卷 / 03期
关键词
Deep immunophenotyping; Multi-parametric flow cytometry; Human whole blood; Gating strategies; Standardization; T-CELL SUBSETS; PHENOTYPIC CHARACTERIZATION; QUALITY-ASSURANCE; B-CELLS; PANEL; LYMPHOCYTE; QUANTIFICATION; VISUALIZATION; EXPRESSION; DRIVEN;
D O I
10.1007/s43657-022-00092-9
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
Immunophenotyping is proving crucial to understanding the role of the immune system in health and disease. High-throughput flow cytometry has been used extensively to reveal changes in immune cell composition and function at the single-cell level. Here, we describe six optimized 11-color flow cytometry panels for deep immunophenotyping of human whole blood. A total of 51 surface antibodies, which are readily available and validated, were selected to identify the key immune cell populations and evaluate their functional state in a single assay. The gating strategies for effective flow cytometry data analysis are included in the protocol. To ensure data reproducibility, we provide detailed procedures in three parts, including (1) instrument characterization and detector gain optimization, (2) antibody titration and sample staining, and (3) data acquisition and quality checks. This standardized approach has been applied to a variety of donors for a better understanding of the complexity of the human immune system.
引用
收藏
页码:309 / 328
页数:20
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