Verhaegen F, Vral A, Seuntjens J, Schipper N W, de Ridder L, Thierens H
Standard Dosimetry Laboratory, Department of Biomedical Physics, Gent, Belgium.
Cytometry. 1994 Oct 1;17(2):119-27. doi: 10.1002/cyto.990170203.
The micronucleus assay in human lymphocytes is, at present, frequently used to assess chromosomal damage caused by ionizing radiation or mutagens. Manual scoring of micronuclei (MN) by trained personnel is very time-consuming, tiring work, and the results depend on subjective interpretation of scoring criteria. More objective scoring can be accomplished only if the test can be automated. Furthermore, an automated system allows scoring of large numbers of cells, thereby increasing the statistical significance of the results. This is of special importance for screening programs for low doses of chromosome-damaging agents. In this paper, the first results of our effort to automate the micronucleus assay with an image-analysis system are represented. The method we used is described in detail, and the results are compared to those of other groups. Our system is able to detect 88% of the binucleated lymphocytes on the slides. The procedure consists of a fully automated localization of binucleated cells and counting of the MN within these cells, followed by a simple and fast manual operation in which the false positives are removed. Preliminary measurements for blood samples irradiated with a dose of 1 Gy X-rays indicate that the automated system can find 89% +/- 12% of the micronuclei within the binucleated cells compared to a manual screening.
目前,人类淋巴细胞微核试验常用于评估电离辐射或诱变剂引起的染色体损伤。由训练有素的人员手动对微核(MN)进行评分是一项非常耗时、累人的工作,而且结果取决于对评分标准的主观解读。只有实现检测自动化,才能进行更客观的评分。此外,自动化系统能够对大量细胞进行评分,从而提高结果的统计学显著性。这对于低剂量染色体损伤剂的筛查项目尤为重要。本文展示了我们使用图像分析系统实现微核试验自动化的初步成果。详细描述了我们所采用的方法,并将结果与其他研究团队的结果进行了比较。我们的系统能够检测出载玻片上88%的双核淋巴细胞。该程序包括对双核细胞进行全自动定位以及对这些细胞内的微核进行计数,随后进行一个简单快速的手动操作以去除假阳性结果。对接受1 Gy X射线照射的血样进行的初步测量表明,与手动筛查相比,自动化系统能够在双核细胞中找到89%±12%的微核。