Automatic analysis of the in vitro micronucleus test on V79 cells

Abstract

The in vitro micronucleus test is a well established test for early screening of new chemical entities in industrial toxicology. For assessing the clastogenic or aneugenic potential of a test compound, micronucleus induction in cells has been shown repeatedly to be a sensitive and specific parameter. As a measure for numerical and structural chromosome aberrations, the in vitro micronucleus test consists of determining the frequency of micronucleated cells in a representative fraction of cells in a culture. So far, manual counting has been the only method for evaluating microscopic V79 Chinese hamster cell preparations. To replace this tedious and time consuming procedure, a fully automatic system for micronucleus scoring in V79 cells by image analysis has been developed and introduced into the routine genotoxicity screening of drug candidates. The comparison of manual and automatic micronucleus analysis showed a high degree of concordance between the results obtained by the two techniques. For concentration series of cyclophosphamide (CP) and ethyl-methanesulphonate (EMS) as test compounds, the frequency of erroneously missed micronuclei through automatic scoring proved to be below 15% in comparison with manual scoring. Generally, false positive micronucleus decisions could be controlled easily by fast and simple relocation of the automatically detected patterns. The possibility to analyze 24 slides within 1 day by fully automatic overnight analysis and the high reproducibility of the results make automatic image processing a powerful tool for the in vitro micronucleus analysis.