Roman D, Locher F, Suter W, Cordier A, Bobadilla M
Preclinical Safety, Toxicology/Pathology, Novartis Pharma AG, Basel, Switzerland.
Environ Mol Mutagen. 1998;32(4):387-96.
Measurement of the frequency of micronuclei induced in cells by ionizing radiation or by chemical treatment is widely used to analyze cytogenetic damage. The microscopic scoring of micronuclei is a tedious and time-consuming procedure. Therefore, attempts have been made to automate micronuclei scoring by means of image analysis or flow cytometry. A new procedure for the flow cytometric analysis of chemically induced micronuclei in V79 Chinese hamster cells has been established in our laboratory. Debris was separated from micronuclei by means of a new gating procedure using area and width fluorescence of the stained suspension of micronuclei and nuclei. In order to test the sensitivity and specificity of this improved method of flow cytometric analysis, five well-known mutagenic compounds were tested. With the new technique, the frequency of micronuclei measured and analyzed corresponded well with results obtained by conventional microscopy. In addition, a large series of negative compounds, and weak, middle, and strong micronuclei inducers, were tested in order to establish criteria for discrimination between genotoxic and nongenotoxic compounds by flow cytometry. This new procedure for flow cytometric detection of micronuclei represents a quick, reliable, and relatively simple method for in vitro micronucleus testing.
通过电离辐射或化学处理诱导细胞产生微核的频率测量被广泛用于分析细胞遗传损伤。微核的显微镜评分是一个繁琐且耗时的过程。因此，人们尝试通过图像分析或流式细胞术实现微核评分的自动化。我们实验室已建立了一种用于V79中国仓鼠细胞中化学诱导微核的流式细胞术分析新方法。通过使用微核和细胞核染色悬浮液的面积和宽度荧光的新门控程序，将碎片与微核分离。为了测试这种改进的流式细胞术分析方法的灵敏度和特异性，对五种著名的诱变化合物进行了测试。采用新技术测量和分析的微核频率与传统显微镜获得的结果非常吻合。此外，还测试了大量阴性化合物以及弱、中、强微核诱导剂，以建立通过流式细胞术区分遗传毒性和非遗传毒性化合物的标准。这种用于流式细胞术检测微核的新方法代表了一种用于体外微核测试的快速、可靠且相对简单的方法。
