Multivariate analysis and list mode processing of murine hemopoietic subpopulations for cytokinetic studies.
Multivariate analysis and list mode processing of murine hemopoietic subpopulations for cytokinetic studies.
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用于细胞因子研究的小鼠造血亚群的多变量分析和列表模式处理。
DOI：
10.1002/cyto.990060608
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
Gray,JW
中科院分区：
文献类型：
--
作者：
Pallavicini,MG;Summers,LJ;Giroud,FJ;Dean,PN;Gray,JW
多变量分析和列表模式的数据处理被用来获得流式细胞术不同的造血亚群的细胞动力学信息。在一个应用中，基于花青染料荧光和正交光散射来区分存活的、未固定的造血亚群;测量Hoechst染料荧光以确定亚群的增殖状态。在另一个应用中，乙醇固定的小鼠骨髓细胞用Hoechst染料、罗丹明缀合的麦胚凝集素（WGA）和荧光素标记的溴脱氧尿苷单克隆抗体进行三重染色。在这两项研究中，所有三个变量的流式细胞术数据均以列表模式采集，存储在磁带上，并在基于计算机的多变量脉冲高度分析仪上通过列表模式软件进行处理。在第一个应用程序中，亚群区分花青染料强度和光散射似乎更相关的细胞谱系和细胞大小比增殖状态。在第二个应用中，WGA亲和力区分了小鼠骨髓S期细胞中的两个亚群，每个亚群中主动掺入溴脱氧尿苷（BrdUrd）。列表模式的数据处理消除了对细胞的常规电子分选的需要，从而有利于区分亚群的表征。在这方面，它是特别有用的流式细胞术不同的，低频率亚群的研究。
Multivariate analyses and list mode data processing were used to obtain cytokinetic information on flow cytometrically distinct hemopoietic subpopulations. In one application, viable, unfixed hemopoietic subpopulations were discriminated on the basis of cyanine dye fluorescence and orthogonal light scatter; Hoechst dye fluorescence was measured to determine the proliferative status of the subpopulations. In another application, ethanol‐fixed mouse bone marrow cells were triply stained with Hoechst dye, rhodamine‐conjugated wheat germ agglutinin (WGA), and a fluorescein‐labeled monoclonal antibody against bromodeoxyuridine. In both of these studies, flow cytometric data for all three variables were acquired in list mode fashion, stored on magnetic tape, and processed by list mode software on a computer based multivariable pulse‐height analyzer. In the first application, subpopulations distinguished by cyanine dye intensity and light scatter appeared to be more related to cell lineage and cell size than proliferative status. In the second application, WGA affinity discriminated two subpopulations in mouse bone marrow S‐phase cells in each subpopulation that actively incorporated bromodeoxyuridine (BrdUrd). List mode data processing obviates the need for routine electronic sorting of cells and thus facilitates characterization of discriminated subpopulations. In this regard, it is particularly useful for the study of flow cytometrically distinct, low frequency subpopulations.