Five-dimensional flow cytometry as a new approach for blood and bone marrow differentials.
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.
Terstappen
L W
LW
Becton Dickinson Monoclonal Center, Inc., Mountain View, California 94043.
Loken
M R
MR
D001773:Blood Cells / Q000166:cytology*; D001792:Blood Platelets / Q000166:cytology; D001854:Bone Marrow Cells; D002051:Burkitt Lymphoma / Q000097:blood; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D004912:Erythrocytes / Q000166:cytology; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007962:Leukocytes / Q000166:cytology; D013921:Thrombocytopenia / Q000097:blood