Lin H C, Sotnikov A V, Fosdick L, Bostick R M, Willett W C
Department of Molecular & Cellular Toxicology, Harvard School of Public Health, Boston, Massachusetts 02115, USA.
Cancer Epidemiol Biomarkers Prev. 1996 Feb;5(2):109-14.
Measurements of proliferating cell nuclear antigen (PCNA) labeling of the large intestinal crypts scored by experienced observers were compared with those generated by computer-assisted image analysis (CAIA). (CAIA was performed at Pathology Expertise, Inc., Newton, MA, by A. V. Sotnikov.) Serial sections (3 microns) of the rectal biopsy specimens from 32 patients were immunostained for PCNA and then counterstained with hematoxylin. The same set of slides and rules for the location/acquisition of complete crypts was used to assess a minimum of ten complete crypts/patient. Each crypt was subdivided longitudinally into five equal compartments. With CAIA, the images were stored digitally, and once the color references were set, the areas occupied by labeled and counterstained nuclei were quantified automatically. The labeling index (LI) was calculated from the PCNA-labeled nuclei area/total nuclei area in CAIA and from the number of labeled cells/total number of cells in visual scoring. The LI of whole crypts averaged 1.04 +/- 0.18 by CAIA and 3.91 +/- 0.46 by the visual method, and the Spearman correlation (rs) between the two methods was 0.89. The different modes of evaluation and color reference selection are likely to have contributed to the differences in the LI ranges observed in the two methods. The high correlation between PCNA quantification by CAIA and visual scoring by experienced technicians indicates that CAIA can reliably rank individual subjects. Thus, measurement of PCNA labeling by CAIA is a practical alternative for evaluating colorectal epithelial cell proliferation.
由经验丰富的观察者对大肠隐窝增殖细胞核抗原（PCNA）标记进行的测量结果，与计算机辅助图像分析（CAIA）生成的结果进行了比较。（CAIA由A. V. Sotnikov在马萨诸塞州牛顿市的Pathology Expertise公司进行。）对32例患者直肠活检标本的连续切片（3微米）进行PCNA免疫染色，然后用苏木精复染。使用同一组载玻片以及获取完整隐窝的位置/规则，对每位患者至少十个完整隐窝进行评估。每个隐窝纵向细分为五个相等的部分。使用CAIA时，图像以数字方式存储，一旦设置了颜色参考，就会自动对标记和复染细胞核所占区域进行量化。标记指数（LI）在CAIA中通过PCNA标记细胞核面积/总细胞核面积计算得出，在视觉评分中通过标记细胞数/细胞总数计算得出。通过CAIA，整个隐窝的LI平均为1.04±0.18，通过视觉方法为3.91±0.46，两种方法之间的Spearman相关性（rs）为0.89。评估方式和颜色参考选择的不同可能导致了两种方法中观察到的LI范围差异。CAIA对PCNA的定量与经验丰富的技术人员的视觉评分之间的高度相关性表明，CAIA可以可靠地对个体进行排名。因此，通过CAIA测量PCNA标记是评估结直肠上皮细胞增殖切实可行的替代方法。
