Five‐dimensional flow cytometry as a new approach for blood and bone marrow differentials
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Open Access
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.Keywords
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