Cope C, Rowe D, Delbridge L, Philips J, Friedlander M
Department of Clinical Oncology, Royal North Shore Hospital, St Leonards, Sydney, South Wales, Australia.
J Clin Pathol. 1991 Feb;44(2):147-51. doi: 10.1136/jcp.44.2.147.
A good correlation (r = 0.94) was obtained between the DNA indices (DI) using flow cytometry and image analysis of nuclei cytospins extracted from paraffin wax embedded tumour sections. Some of the limitations and problems associated with image analysis which came to light included an unacceptably high coefficient of variation (CV) and a "left-shift" in the DI in most DNA histograms obtained when using image analysis of 5 microns sections. In contrast, the DNA histograms generated using image analysis of cytospun nuclei from paraffin wax blocks were of good quality and similar to those obtained using flow cytometry. Variability in Feulgen staining was common and an important source of error despite rigorous control of the staining technique. This could be overcome by using internal controls such as fibroblasts rather than external controls (rat hepatocytes) to determine the diploid DI with image analysis. A thorough understanding and appreciation of the methodological problems associated with image analysis and flow cytometric determination of DNA content is required before these methods find widespread clinical application.
采用流式细胞术测定的DNA指数（DI）与从石蜡包埋肿瘤切片中提取的细胞核涂片进行图像分析所得到的结果之间具有良好的相关性（r = 0.94）。图像分析中出现的一些局限性和问题包括变异系数（CV）高得难以接受，以及在对5微米切片进行图像分析时，大多数DNA直方图中的DI出现“左移”。相比之下，对石蜡块中细胞核涂片进行图像分析所生成的DNA直方图质量良好，与采用流式细胞术得到的结果相似。尽管对染色技术进行了严格控制，但福尔根染色的变异性仍然常见且是一个重要的误差来源。通过使用成纤维细胞等内部对照而非外部对照（大鼠肝细胞）来通过图像分析确定二倍体DI，可以克服这一问题。在这些方法得到广泛临床应用之前，需要对与图像分析和DNA含量流式细胞术测定相关的方法学问题有透彻的理解和认识。
