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Updated: Jul 2, 2026
Enumeration of Major Peripheral Blood Leukocyte Populations for Multicenter Clinical Trials Using a Whole Blood Phenotyping Assay
Published on: September 16, 2012
Immunophenotypic identification, enumeration, and characterization of human peripheral blood dendritic cells and
Insights
Identifying dendritic cells (DCs) and their subsets in blood is challenging due to low numbers and lack of specific markers. This study provides optimized flow cytometry protocols for accurate DC identification and enumeration.
Area of Science:
- Immunology
- Cell Biology
Background:
- Dendritic cells (DCs) are crucial immune cells, but their low frequency and heterogeneity complicate analysis.
- Specific markers for sensitive and precise DC identification are lacking, hindering research.
Purpose of the Study:
- To present optimized flow cytometry protocols for identifying and enumerating dendritic cells and their subsets in peripheral blood.
- To address challenges in DC analysis, including low cell frequency and lack of specific markers.
Main Methods:
- Utilized a two-step acquisition procedure in flow cytometry to increase cell analysis numbers.
- Developed multiparametric gating strategies for specific identification of DC subsets.
- Employed erythrocyte-lysed whole peripheral blood for sample preparation.
Main Results:
- Established protocols for the identification and enumeration of all dendritic cells.
- Successfully identified and enumerated various dendritic cell subsets.
- Demonstrated effective multiparametric gating strategies for precise DC analysis.
Conclusions:
- The presented flow cytometry protocols enable sensitive and specific identification and enumeration of dendritic cells and their subsets.
- These methods overcome challenges associated with low DC frequency and marker specificity.
- The protocols facilitate deeper understanding of dendritic cell populations in immunological studies.
Abstract:
Study of dendritic cells is complicated by their low frequency and the lack of specific markers for sensitive and specific identification. Furthermore, these cells are not a homogeneous population, but contain a variety of subsets. Flow cytometry has well-established applications for the analysis of cells present at low frequencies; the use of a two-step acquisition procedure increases the number of cells for analysis. This unit presents protocols for the identification and enumeration of dendritic cells and cell subsets in erythrocyte-lysed whole peripheral blood. Special attention is paid to the combinations of reagents used for the identification of all dendritic cells as well as each subset, and on the multiparametric gating strategies for specific analysis.
