Ottesen G L, Christensen I J, Larsen J K, Hansen B, Andersen J A
Institute of Pathology, Odense University Hospital, Denmark.
Cytometry. 1996 Mar 15;26(1):65-8. doi: 10.1002/(SICI)1097-0320(19960315)26:1<65::AID-CYTO10>3.0.CO;2-Q.
Flow cytometric DNA analysis was performed on unfixed frozen samples from 56 breast cancer patients. From each patient, two samples were analyzed. The only difference in the handling of the paired samples was the mechanical disaggregation of one sample by fine-needle aspiration compared to an automated mechanical disaggregation method (Medimachine) of the other sample. With the two methods for tissue disaggregation, the same resolution of the DNA histograms was obtained, indicated by median coefficients of variation (CV) of 1.5% for the DNA diploid G1 peaks. Also, the frequencies of DNA diploid and aneuploid cases as well as the fractions of DNA aneuploid cells were comparable. This indicates that the two methods did not differ in ability to detect DNA aneuploid tumor clones. Automated mechanical disaggregation resulted in DNA histograms with significantly less debris and with lower S-phase fractions. In practice, the procedure of automated mechanical disaggregation was rapid, easy, and safer because of minimal handling of the unfixed tissue compared to the fine-needle aspiration.
对56例乳腺癌患者未固定的冷冻样本进行了流式细胞术DNA分析。每位患者分析两个样本。配对样本处理的唯一区别在于，一个样本通过细针穿刺进行机械解离，另一个样本采用自动机械解离方法（Medimachine）。采用这两种组织解离方法，DNA直方图的分辨率相同，DNA二倍体G1峰的中位变异系数（CV）为1.5%，表明了这一点。此外，DNA二倍体和非整倍体病例的频率以及DNA非整倍体细胞的比例相当。这表明这两种方法在检测DNA非整倍体肿瘤克隆的能力上没有差异。自动机械解离产生的DNA直方图碎片明显更少，S期比例更低。在实际操作中，与细针穿刺相比，自动机械解离过程快速、简便且更安全，因为对未固定组织的处理最少。
