Ornberg R L
Pharmacia Corporation, St Louis, Missouri 63198, USA.
J Histochem Cytochem. 2001 Aug;49(8):1059-60. doi: 10.1177/002215540104900815.
Cell proliferation and apoptosis indices are important indicators for the prognosis and treatment of a variety of cancers. A method is described using differential absorption color image analysis to measure proliferation and apoptosis in tumor sections using BrdU (5' bromodeoxyuridine) incorporation and immunohistochemistry and terminal deoxytransferase nick end-labeling (TUNEL). Nuclei were labeled with streptavidin-peroxidase-diaminobenzidine (DAB) secondary detection. The differential absorption method uses a computer-controlled microscope equipped with a tunable filter and digital camera to take advantage of the spectral differences of stained objects of interest. Images collected at defined wavelengths are divided and scaled to form ratio images in which the hematoxylin- or DAB-stained nuclei have intensity ranges far above those of surrounding structures. Using brightness thresholding followed by selection based on nuclear size and shape parameters, binary images were formed of the BrdU/apoptotic-positive tumor and all the tumor nuclei for subsequent counting and calculations of proliferation and apoptotic indices.
细胞增殖和凋亡指数是多种癌症预后和治疗的重要指标。本文描述了一种使用差分吸收彩色图像分析的方法，通过5-溴脱氧尿苷（BrdU）掺入、免疫组织化学和末端脱氧核苷酸转移酶介导的缺口末端标记法（TUNEL）来测量肿瘤切片中的增殖和凋亡情况。细胞核用链霉亲和素-过氧化物酶-二氨基联苯胺（DAB）进行二次检测标记。差分吸收法使用配备可调滤光片和数码相机的计算机控制显微镜，利用感兴趣的染色物体的光谱差异。在特定波长下采集的图像进行分割和缩放，以形成比率图像，其中苏木精或DAB染色的细胞核的强度范围远高于周围结构。通过亮度阈值处理，然后根据核大小和形状参数进行选择，形成BrdU/凋亡阳性肿瘤和所有肿瘤细胞核的二值图像，用于后续增殖和凋亡指数的计数和计算。
