Ottesen G L, Christensen I J, Larsen J K, Larsen J, Christiansen J, Baldetorp B, Linden T, Hansen B, Andersen J A
Center for Breast Cancer Research, Institute of Pathology, Odense University Hospital, Denmark.
Anal Quant Cytol Histol. 1997 Oct;19(5):413-22.
To form a methodologic basis for DNA analysis of ductal carcinoma in situ (DCIS) and invasive carcinoma (IC) of the breast, including very small lesions, by comparison of flow cytometric (FCM) and image cytometric (ICM) methods for DNA quantitation.
The material consisted of 41 DCIS lesions and 26 ICs. FCM DNA analysis of unfixed, frozen samples were compared to (1) FCM of formalin-fixed, paraffin-embedded tissue; (2) ICM of imprints; and (3) ICM of paraffin-embedded tissue sections.
FCM of unfixed tissue showed higher DNA measurement precision and a higher number of DNA nondiploid clones as compared to the other three methods. For the classification of DNA diploid/nondiploid cases, high concordance rates were found between the methods. Discordant cases were predominantly DNA neardiploid by FCM of unfixed tissue but DNA diploid by the other methods. The reproducibility of the DNA index (DI) was best in the interval 1.2 < DI < or = 2.2; it was 74% for FCM of fixed tissue and 79% for ICM of imprints. Clones with DI > 3 were found almost exclusively by ICM of imprints. For ICM of tissue sections, DI could not be reliably estimated. By ICM, contrary to FCM, a combined DNA diploid and nondiploid pattern was found frequently.
Each of the methods has its own advantages and limitations. If possible, FCM should be combined with ICM. FCM of unfixed tissue is superior to the other methods with respect to precise DI estimation. Alternatively, FCM of fixed tissue and ICM of imprints may both give a reliable estimate of DI. ICM of tissue sections can discriminate DNA diploid from nondiploid clones, except for neardiploid subpopulations, and permits the analysis of very small lesions.
通过比较流式细胞术（FCM）和图像细胞术（ICM）进行DNA定量分析的方法，为乳腺导管原位癌（DCIS）和浸润性癌（IC）（包括非常小的病变）的DNA分析建立方法学基础。
材料包括41例DCIS病变和26例IC。将未固定的冰冻样品的FCM DNA分析与以下方法进行比较：（1）福尔马林固定、石蜡包埋组织的FCM；（2）印片的ICM；（3）石蜡包埋组织切片的ICM。
与其他三种方法相比，未固定组织的FCM显示出更高的DNA测量精度和更多的DNA非二倍体克隆。对于DNA二倍体/非二倍体病例的分类，各方法之间的一致性率较高。不一致的病例主要是未固定组织的FCM显示为DNA近二倍体，但其他方法显示为DNA二倍体。DNA指数（DI）的可重复性在1.2＜DI≤2.2区间最佳；固定组织的FCM为74%，印片的ICM为79%。DI＞3的克隆几乎仅在印片的ICM中发现。对于组织切片的ICM，DI无法可靠估计。与FCM相反，通过ICM经常发现DNA二倍体和非二倍体的组合模式。
每种方法都有其自身的优点和局限性。如果可能，FCM应与ICM相结合。就精确估计DI而言，未固定组织的FCM优于其他方法。或者，固定组织的FCM和印片的ICM都可以对DI进行可靠估计。组织切片的ICM可以区分DNA二倍体和非二倍体克隆，但近二倍体亚群除外，并允许对非常小的病变进行分析。
