Gratzner H G, Young I T, Sher S E
J Histochem Cytochem. 1979 Jan;27(1):496-9. doi: 10.1177/27.1.374614.
Antibodies specific for 5-bromodeoxyuridine (BrdUrd) can be used to measure labeling indices in an automated system by image analysis. The antibody, used with an indirect immunoperoxidase technique, will detect de novo DNA synthesis subsequent to growing the cells for various time intervals in 5-bromodeoxyuridine-containing medium. Asynchronously growing CHO cells were pulsed with 3H-5-bromodeoxyuridine, fixed, denatured and then stained with anti-bromouridine antiserum. Peroxidase-coupled goat anti-rabbit IgG was used as the secondary antibody, and slides were stained with diaminobenzidine. Cells which are positive display a reticular pattern indicative of replicating chromatin. "Labeling indices" were generated by scanning the nuclei by TV image analysis. The percentage of labeled cells by the immunocytochemical technique correlates well with that found by autoradiography. Some of the applications of this automated method include cell kinetics and analysis of S-phase by pattern recognition technique.
针对5-溴脱氧尿苷（BrdUrd）的特异性抗体可用于通过图像分析在自动化系统中测量标记指数。该抗体与间接免疫过氧化物酶技术一起使用，将在含5-溴脱氧尿苷的培养基中培养细胞不同时间间隔后检测新生DNA合成。将异步生长的中国仓鼠卵巢（CHO）细胞用3H-5-溴脱氧尿苷脉冲处理，固定、变性，然后用抗溴尿苷抗血清染色。过氧化物酶偶联的山羊抗兔IgG用作二抗，玻片用二氨基联苯胺染色。呈阳性的细胞显示出网状模式，表明染色质正在复制。通过电视图像分析扫描细胞核生成“标记指数”。免疫细胞化学技术标记细胞的百分比与放射自显影法得到的结果相关性良好。这种自动化方法的一些应用包括细胞动力学和通过模式识别技术分析S期。
