Automatic analysis of the micronucleus test in primary human lymphocytes using image analysis

This study assessed the clastogenic or aneugenic potential of a test compound, micronucleus induction in cells as a sensitive and a specific parameter. The ROBIAS (Robotic Image Analytical System) for both automatic cytotoxicity assessment and micronucleus detection in human lymphocytes was developed at Novartis where the assay has been used to validate possible results obtained in the micronucleus test(MNT) in TK6 cells, which serves as the primary screening system for genotoxicity profiling in early drug development. In addition, the in vitro MNT has become an accepted alternative to support clinical studies and will be used for regulatory purposes as well. The comparison of visual with automatic analysis results showed a high degree of concordance for 25 independent experiments conducted for the profiling of 12 compounds. For concentration series of cyclophosphamide and carbendazim, a very good correlation between automatic and visual analysis by two examiners could be established, both for the relative division index used as cytotoxicity parameter, as well as for micronuclei scoring in mono- and binucleated cells. Generally, false-possible micronucleus decisions could be controlled by fast and simple relocation of the automatically detected patterns. The possibility to analyse 24 slides within 65h by automatic analysis over the weekend and the high reproducibility of the results make automatic image processing a powerful tool for the micronucleus analysis in primary human lymphocytes. The author concluded the automated slide analysis for the MNT in human lymphocytes complements the portfolio of image analytical applications on ROBIAS, which is supporting various assays at Novartis.

Authors: Frieauff, W.; Martus, H. J.; Suter, W.; Elhajouji, A. ;Full Source: Mutagenesis 2013, 28(1), 15-23 (English) ;