Gonchoroff N J, Ryan J J, Kimlinger T K, Witzig T E, Greipp P R, Meyer J S, Katzmann J A
Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota 55905.
Cytometry. 1990;11(5):642-6. doi: 10.1002/cyto.990110513.
Since the publication of paraffin block extraction procedures, flow cytometric analysis of DNA ploidy and S-phase of tumor specimens has been widely applied. DNA aneuploidy, DNA tetraploid (elevated G2/M), and elevated S-phase are clinically significant in some tumor systems. True DNA tetraploid cell lines will contain a large 4c population and perhaps an 8c population; samples with cell aggregates will also contain a 6c population. Microscopic examination of samples having a 6c peak revealed nuclei with adhering debris and doublets, triplets, and larger nuclear aggregates. After sonication, a uniform suspension of single nuclei without adherent debris was seen. In addition to reducing the percent of G2/M cells, sonication also reduced S-phase percent such that it was closer to the bromodeoxyuridine labeling index. The DNA ploidy classification of specimens was also compared pre- and post-sonication. Four of 96 breast cancer samples changed classification; all were specimens in which the histogram became cleaner and a small DNA aneuploid peak became apparent after sonication.
自从石蜡块提取程序发表以来，肿瘤标本的DNA倍性和S期的流式细胞术分析已被广泛应用。DNA非整倍体、DNA四倍体（G2/M期升高）和S期升高在某些肿瘤系统中具有临床意义。真正的DNA四倍体细胞系将包含大量的4c群体，可能还包含一个8c群体；有细胞聚集体的样本也将包含一个6c群体。对具有6c峰的样本进行显微镜检查，发现细胞核带有附着的碎片以及双核、三核和更大的核聚集体。超声处理后，观察到单核的均匀悬浮液，没有附着的碎片。除了降低G2/M期细胞的百分比外，超声处理还降低了S期百分比，使其更接近溴脱氧尿苷标记指数。还比较了超声处理前后标本的DNA倍性分类。96个乳腺癌样本中有4个改变了分类；所有这些标本在超声处理后直方图变得更清晰，并且出现了一个小的DNA非整倍体峰。
