Asano N, Katsuma Y, Tamura H, Higashikuni N, Hayashi M
Toxicological Research Division, Nitto Denko Corporation, 1-1-2, Shimohozumi, Ibaraki, Osaka 567-8680, Japan.
Mutat Res. 1998 Aug 3;404(1-2):149-54. doi: 10.1016/s0027-5107(98)00108-0.
We developed an automated image analysis system to obtain objective data for the rodent peripheral blood micronucleus assay with acridine orange (AO) supravital staining. The system was able to identify micronucleated reticulocytes (MNRETs) and to evaluate inhibition of bone marrow cell proliferation by measuring the reticular area of reticulocytes (RETs). We also developed automated equipment to produce homogeneous acridine orange-coated glass slides. This study was designed to compare automated scoring with manual scoring using 4 model clastogens and 2 mouse strains. The MNRET incidence induced by each clastogen was similar for automated and manual scoring, and there was good correlation (r=0.92) between the methods. In addition, an index of bone marrow toxicity based on the reticular area of RETs was compared to the conventional index (% of polychromatic erythrocytes (PCE) to total erythrocytes; PCE ratio) and was similar. The results indicated that our technique for computer-assisted image analysis for the micronucleus assay with AO supravitally stained peripheral blood RETs was comparable to conventional microscopic scoring, and it was superior in objectivity and statistical power.
我们开发了一种自动化图像分析系统，用于通过吖啶橙（AO）活体染色获取啮齿动物外周血微核试验的客观数据。该系统能够识别微核网织红细胞（MNRET），并通过测量网织红细胞（RET）的网状面积来评估骨髓细胞增殖的抑制情况。我们还开发了用于生产均匀涂有吖啶橙的载玻片的自动化设备。本研究旨在使用4种模型致裂剂和2种小鼠品系比较自动评分和手动评分。每种致裂剂诱导的MNRET发生率在自动评分和手动评分中相似，并且两种方法之间具有良好的相关性（r = 0.92）。此外，将基于RET网状面积的骨髓毒性指数与传统指数（多染性红细胞（PCE）占总红细胞的百分比；PCE比率）进行比较，结果相似。结果表明，我们用于AO活体染色外周血RET微核试验的计算机辅助图像分析技术与传统显微镜评分相当，并且在客观性和统计效力方面更具优势。
