Immunophenotypic identification, enumeration, and characterization of human peripheral blood dendritic cells and...

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.