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

作　　者：Jian Gao Yali Luo Helian Li Yiran Zhao Jialin Zhao Xuling Han Jingxuan Han Huiqin Lin Feng Qian 

机构地区：[1]State Key Laboratory of Genetic Engineering,Shanghai Public Health Clinical Center,Human Phenome Institute,Zhangjiang Fudan International Innovation Center and School of Life Sciences,Fudan University,Shanghai 200438,China [2]Ministry of Education Key Laboratory of Contemporary Anthropology,School of Life Sciences,Fudan University,Shanghai 200438,China [3]Institute of Immunophenome,International Human Phenome Institutes(Shanghai),Shanghai 200433,China

出　　处：《Phenomics》2023年第3期309-328,共20页表型组学（英文）

基　　金：supported by the National Key Research and Development Program of China(2021YFA1301000);Shanghai Municipal Science and Technology Major Project(Grant No.2017SHZDZX01).

摘　　要：Immunophenotyping is proving crucial to understanding the role of the immune system in health and disease.High-through-put 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.

关 键 词：Deep immunophenotyping Multi-parametric flow cytometry Human whole blood Gating strategies Standardization 

分 类 号：R392[医药卫生—免疫学]