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Automatic Analysis of the Micronucleus Test in Primary Human Lymphocytes Using Image Analysis

Frieauff, Wilfried, Martus, Hans-Joerg, Suter, Willi and Elhajouji, Azeddine (2012) Automatic Analysis of the Micronucleus Test in Primary Human Lymphocytes Using Image Analysis. Mutagenesis.

Abstract

The in vitro micronucleus test (MNT) 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. Various automated systems to replace the tedious and time consuming manual slide analysis procedure have been described. Flow cytrometric approaches have been discussed elsewhere. The ROBIAS image analysis system for both automatic cytotoxicity assessment and highly sensitive micronucleus detection in primary human lymphocytes was developed at Novartis, where the assay is used as to confirm positive results obtained in the MNT in TK6 cells which serves as the primary screening system for genotoxicity profiling in early drug development. The comparison of manual with automatic analysis results showed a high degree of concordance for 27 independent experiments conducted for profiling of 12 compounds. For concentration series of Cyclophosphamide (CP) and Carbendazim (MBC), a very good correlation between automatic and manual analysis could be established, both for the relative division index used as cytotoxicity parameter, and for MN scoring in mono- and bi-nucleated cells. Generally, false positive micronucleus decisions could be controlled by fast and simple relocation of the automatically detected patterns. The possibility to analyze 24 slides within 65 hours by fully 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 automated slide analysis for the MNT in human lymphocytes complements the portfolio of image analysis applications on ROBIAS supporting various assays in genetic toxicology and other biomedical areas.

Item Type: Article
Keywords: in vitro, micronucleus test, human lymphocytes, image analysis
Date Deposited: 13 Oct 2015 13:16
Last Modified: 13 Oct 2015 13:16
URI: https://oak.novartis.com/id/eprint/3307

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