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  • Title: Simultaneous identification of eight leucocyte subsets of human peripheral blood using three-colour immunofluorescence flow cytometric analysis.
    Author: Carayon P, Bord A, Raymond M.
    Journal: J Immunol Methods; 1991 Apr 25; 138(2):257-64. PubMed ID: 2033277.
    Abstract:
    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.
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