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  • Title: Five-dimensional flow cytometry as a new approach for blood and bone marrow differentials.
    Author: Terstappen LW, Loken MR.
    Journal: Cytometry; 1988 Nov; 9(6):548-56. PubMed ID: 3208621.
    Abstract:
    We have used five independent variables on a flow cytometer to discriminate and to quantify the cellular components within both blood and bone marrow aspirates. The signals were stored in list mode by which a five-dimensional space was created. The cells--differentiated into: 1) erythrocytes, 2) reticulocytes, 3) nucleated erythroid cells, 4) platelets, 5) lymphocytes, 6) monocytes, 7) neutrophils, 8) eosinophils, and 9) immature leukocytes--had to meet unique criteria with regard to their characteristics in the created five-dimensional space in order to be classified in a specific cell category. Forward and orthogonal light-scattering signals were matched with three fluorescence variables to obtain discrimination without necessitating erythrocyte lysis. Thiazole orange (binding predominantly to RNA) and LDS-751 (principally detecting DNA) were used to differentiate erythrocytes, platelets, reticulocytes, and nucleated cells. A monoclonal antibody, CD45, conjugated with phycoerythrin, was used to aid in discriminating between lineages of nucleated cells.
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