Greimers R, Rongy A M, Schaaf-Lafontaine N, Boniver J
Laboratory of Pathologic Anatomy, University Hospital of Liege, Belgium.
Cytometry. 1991;12(6):570-8. doi: 10.1002/cyto.990120614.
The CUBIC program displays three-dimensional colored dot plots of flow cytometric trivariate data collected by unmodified commercial instruments (FACScan flow cytometer, FACS 440 cell sorter). Assuming a bimodal distribution of the fluorescence intensity of the cells, the eight theoretical subpopulations involved in a three-color fluorescence histogram are clearly localized in the 3-D space by colored dots that are clustered near each corner of a cubic frame. Rotation, tilting, and zoom functions are available. Table look-up is not needed. CUBIC was illustrated by two experiments: 1) three-color immunofluorescence of antigens on human lymphocytes using monoclonal antibodies conjugated either to fluorescein (FITC), to R-phycoerythrin (PE), or to biotin revealed by a streptavidin coupled to a PE-Texas red tandem conjugate (TC); 2) two-color immunofluorescence of CD4 and CD8 antigens on thymocytes of healthy or preleukemic mice correlated to the DNA content quantified by 7-amino-actinomycin D (7-AAD). The three fluorescences were excited by a single argon-ion laser emitting at 488 nm.
CUBIC程序可显示由未改装的商用仪器（FACScan流式细胞仪、FACS 440细胞分选仪）收集的流式细胞术三变量数据的三维彩色散点图。假设细胞荧光强度呈双峰分布，三色荧光直方图中涉及的八个理论亚群通过彩色点清晰地定位在三维空间中，这些点聚集在立方框架的每个角附近。提供旋转、倾斜和缩放功能。无需查表。通过两个实验对CUBIC进行了说明：1）使用与荧光素（FITC）、藻红蛋白（PE）或生物素偶联的单克隆抗体对人淋巴细胞上的抗原进行三色免疫荧光，生物素由与PE-德克萨斯红串联共轭物（TC）偶联的链霉亲和素显示；2）健康或白血病前期小鼠胸腺细胞上CD4和CD8抗原的双色免疫荧光与通过7-氨基放线菌素D（7-AAD）定量的DNA含量相关。三种荧光由发射波长为488 nm的单氩离子激光激发。
