Terstappen L W, Loken M R
Becton Dickinson Monoclonal Center, Inc., Mountain View, California 94043.
Cytometry. 1988 Nov;9(6):548-56. doi: 10.1002/cyto.990090607.
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.
我们在流式细胞仪上使用了五个独立变量，以区分和量化血液及骨髓抽吸物中的细胞成分。信号以列表模式存储，由此创建了一个五维空间。这些细胞分为：1）红细胞，2）网织红细胞，3）有核红细胞，4）血小板，5）淋巴细胞，6）单核细胞，7）中性粒细胞，8）嗜酸性粒细胞，以及9）未成熟白细胞，它们必须在创建的五维空间中满足关于其特征的独特标准，才能被归类到特定的细胞类别中。前向和正交光散射信号与三个荧光变量相匹配，以实现无需红细胞裂解的鉴别。噻唑橙（主要与RNA结合）和LDS - 751（主要检测DNA）用于区分红细胞、血小板、网织红细胞和有核细胞。一种与藻红蛋白偶联的单克隆抗体CD45用于辅助鉴别有核细胞的谱系。
