Simultaneous identification of eight leucocyte subsets of human peripheral blood using three-colour immunofluorescence flow cytometric analysis.
The full analytical potential of flow cytometry has been exploited in order to identify a maximum of human peripheral blood leucocyte subsets in a single tube. For this purpose a mixture composed of six different monoclonal antibodies directly coupled to FITC or phycoerythrin or indirectly linked to the DuoChrome reagent via the biotin/streptavidin system was used. This combination of monoclonal antibodies and fluorochromes offered the possibility of simultaneously determining eight leucocyte subsets of human peripheral blood using a single laser beam. These subsets were T4 cells, bright and dim T8 cells, T null (CD3+ CD4- CD8-) cells, NK cells and B cells, polymorphonuclear cells and monocytes. This method is very easy to perform, can be applied to samples from other lymphoid tissues such as tonsils and is particularly useful when starting with a low number of cells.
Carayon
P
P
Immunology Department, Sanofi Recherche, Montpellier, France.
Bord
A
A
Raymond
M
M
D015703:Antigens, CD / Q000276:immunology; D016175:B-Lymphocyte Subsets / Q000276:immunology*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D016130:Immunophenotyping; D007694:Killer Cells, Natural / Q000276:immunology; D009000:Monocytes / Q000276:immunology; D009504:Neutrophils / Q000276:immunology; D016176:T-Lymphocyte Subsets / Q000276:immunology*
D015703:Antigens, CD