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Updated: Jul 26, 2025
A Semi-automated Approach to Preparing Antibody Cocktails for Immunophenotypic Analysis of Human Peripheral Blood
Published on: February 8, 2016
Deep Immunophenotyping of Human Whole Blood by Standardized Multi-parametric Flow Cytometry Analyses
Jian Gao1,2, Yali Luo1,2, Helian Li1,2
1State 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.
Abstract:
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.
Supplementary Information:
The online version contains supplementary material available at 10.1007/s43657-022-00092-9.
