Rew D A, Reeve L J, Wilson G D
University Department of Surgery, University of Leicester, United Kingdom.
Cytometry. 1998 Nov 1;33(3):355-61. doi: 10.1002/(sici)1097-0320(19981101)33:3<355::aid-cyto10>3.0.co;2-s.
The introduction of the laser scanning cytometer offers new capabilities in cell proliferation research, through its capacity for validation of each and every cell event through direct visualization on the microscope slide. In this study, we report a direct comparison of proliferation data derived from flow and laser scanning cytometry of human tumor nuclei labeled in vivo with bromodeoxyuridine (BrdUrd). Nuclear suspensions from 19 invasive ductal breast carcinomas and 12 gastric adenocarcinomas were prepared and analysed for BrdUrd uptake and DNA content. Specimens were analysed using a FACScan and then prepared on cytocentrifuge preparations for laser scanning cytometry. DNA index, labeling index (LI), duration of S-phase (Ts) and potential doubling time (Tpot) were calculated using standard procedures. There was an excellent correlation between the two techniques in the calculation of DNA index (R = 0.983, P > 0.0001) and LI (R = 0.924, P > 0.0001). The Ts proved more problematical (R = 0.448, P = 0.0115) but the Tpot showed closer agreement (R = 0.851, P > 0.0001) as the LI was the dominant determinant of Tpot. No single parameter could be identified as the major source of variation between the two techniques. We conclude that the laser scanning cytometer produces data equivalent to that obtained by flow cytometry.
激光扫描细胞仪的引入为细胞增殖研究提供了新的功能，因为它能够通过在显微镜载玻片上直接观察来验证每一个细胞事件。在本研究中，我们报告了对体内用溴脱氧尿苷（BrdUrd）标记的人肿瘤细胞核进行流式细胞术和激光扫描细胞术所获得的增殖数据的直接比较。制备了来自19例浸润性导管乳腺癌和12例胃腺癌的核悬液，并分析了BrdUrd摄取和DNA含量。使用FACScan对标本进行分析，然后制备细胞离心涂片用于激光扫描细胞术。使用标准程序计算DNA指数、标记指数（LI）、S期持续时间（Ts）和潜在倍增时间（Tpot）。在计算DNA指数（R = 0.983，P > 0.0001）和LI（R = 0.924，P > 0.0001）时，两种技术之间存在极好的相关性。Ts的情况更成问题（R = 0.448，P = 0.0115），但由于LI是Tpot的主要决定因素，Tpot显示出更接近的一致性（R = 0.851，P > 0.0001）。没有一个单一参数可以被确定为两种技术之间差异的主要来源。我们得出结论，激光扫描细胞仪产生的数据与流式细胞术获得的数据相当。
