Evaluation of high-throughput screening for in vitro micronucleus test using fluorescence-based cell imaging

Abstract

The in vitro micronucleus (MN) test is an important component of genotoxicity screening and is used as an alternative to the in vitro chromosome aberration (CA) test. As the MN assay is more practical and simpler to use than the CA test, it is being applied as a high-throughput screening (HTS) assay. Therefore, we conducted a validation study of the MN test employing a confocal imaging plate reader, the IN Cell Analyzer 1000. We evaluated 30 chemicals, including clastogens and aneugens, using Chinese hamster lung cells (CHL/IU) seeded in 96-well microplates. The microplates were stained with Hoechst 33342 and CellMask Red for automated analysis, and MN were identified and counted automatically in fluorescence images. The MN test results for 30 chemicals obtained with this image analysis system, using the IN Cell Analyzer, were highly consistent with reference data for the in vitro MN test and CA test data obtained by microscopic analysis. In conclusion, this HTS assay for detecting MN offers high efficiency and accuracy in the early stages of chemical development.