Truong K, Vielh P, Malfoy B, Klijanienko J, Dutrillaux B, Bourgeois C A
Cytogénétique moléculaire et Oncologie, Institut Curie, Paris, France.
Cancer. 1998 Oct 25;84(5):309-16.
Automated image cytometry can allow concurrent quantification of several parameters in each individual cell within a population, opening new possibilities for diagnosis and prognosis. In this study, the authors investigated the capacity of this method for performing a bivariate analysis of DNA ploidy and synthesis in fine-needle samplings obtained without aspiration from breast tumors.
Samplings from 25 unselected cases of ductal infiltrative breast adenocarcinoma and 2 cases of fibroadenoma were analyzed. For each case, 3-5 slides (containing approximately 1000 cells each) were quantified to assess experimental precision. Ploidy was determined by fluorescent staining of DNA using 4,6-diamidino-2-phenylindole (DAPI). Contaminating lymphocytes were taken as internal controls to calculate DNA indices. DNA synthesis was analyzed by immunofluorescent detection of 5-bromodeoxyuridine (BrdU) incorporation. Measurements were compared with flow cytometric data obtained from the same patients.
Relative error in determination of DNA indices was generally below 5%. Determination of proliferation indices were more variable, with a mean relative error of 25%. Two different populations of BrdU positive cells were detected systematically, one in the diploid and another in the aneuploid fraction. For both cytometric methods, DNA indices were similar in all 27 cases, whereas BrdU labeling indices showed no significant correlation in 13 cases. The remaining cases were not comparable due to lack of flow cytometric data. Labeling indices obtained by image cytometry did not reveal any significant correlation with Scarff-Bloom-Richardson grading or clinical staging.
Automated image cytometry allows concurrent measurement of ploidy and cell proliferation within individual breast carcinoma cells. Statistical reliability can be reached with a relative small number of cells (1000), which is crucial for samples in which the cell number is too low for flow cytometry analysis. Visual control for artifact elimination and better characterization of cell populations makes this a powerful tool for tumor cell investigation. Automated image cytometry allows the obtainment of valuable prognostic parameters of traditional flow cytometry with the relatively small number of cells obtained in aspiration procedures.
自动图像细胞术能够同时对群体中每个细胞的多个参数进行定量分析，为诊断和预后评估开辟了新的可能性。在本研究中，作者探讨了该方法对未经抽吸获取的乳腺肿瘤细针采样进行DNA倍体和合成的双变量分析的能力。
对25例未经选择的导管浸润性乳腺腺癌病例和2例纤维腺瘤病例的采样进行分析。对于每个病例，对3 - 5张载玻片（每张载玻片约含1000个细胞）进行定量分析以评估实验精度。使用4,6 - 二脒基 - 2 - 苯基吲哚（DAPI）通过DNA荧光染色确定倍体。将污染的淋巴细胞作为内部对照来计算DNA指数。通过免疫荧光检测5 - 溴脱氧尿苷（BrdU）掺入来分析DNA合成。将测量结果与从同一患者获得的流式细胞术数据进行比较。
DNA指数测定的相对误差一般低于5%。增殖指数的测定变异性更大，平均相对误差为25%。系统地检测到两个不同的BrdU阳性细胞群体，一个在二倍体部分，另一个在非整倍体部分。对于两种细胞术方法，所有27例病例的DNA指数相似，而13例病例中BrdU标记指数无显著相关性。其余病例由于缺乏流式细胞术数据无法进行比较。通过图像细胞术获得的标记指数与斯卡夫 - 布鲁姆 - 理查森分级或临床分期均无显著相关性。
自动图像细胞术能够同时测量单个乳腺癌细胞内的倍体和细胞增殖情况。相对少量的细胞（1000个）就能达到统计可靠性，这对于细胞数量过少无法进行流式细胞术分析的样本至关重要。通过视觉控制消除假象并更好地表征细胞群体，使其成为肿瘤细胞研究的有力工具。自动图像细胞术能够利用在抽吸程序中获得的相对少量细胞获取传统流式细胞术的有价值预后参数。
