Automated image analysis of micronuclei by IMSTAR for biomonitoring

Abstract

For many years, the analysis of micronuclei (MN) has been successfully applied to human biomonitoring of in vivo genotoxin exposure and provides a sensitive and relatively easy methodology to assess genomic instability. However, there is a need for automation of MN analysis for rapid, more reliable and non-subjective MN detection. In this review, we evaluate the application of automated image analysis of the in vitro cytokinesis-block MN assay on human lymphocytes for human biomonitoring, starting with the requirements that should be fulfilled by a valid and efficient image analysis system. Considering these prerequisites, we compare the automated facility developed in the framework of the European Union-project NewGeneris with other already published systems for automated scoring of MN. Although the automated scoring of MN is now put into place, extension to other cytome assay end points such as apoptosis, necrosis, nuclear buds and nucleoplasmic bridges would greatly enhance the specificity and sensitivity of future biomonitoring studies. Inclusion of these end points would also allow an automated approach to in vitro genotoxicity testing. In addition, automated scoring of the MN assay in exfoliated buccal cells would be very beneficial for large-scale biomonitoring studies, as cells can be collected in a minimally invasive manner.

Laboratory Intercomparison of the Cytokinesis-Block Micronucleus Assay

“…Several automated systems are available on the market and have been tested by human monitoring (28,(31)(32)(33)(34). In our study the scoring system of MetaSystems (Altlussheim, Germany) was utilized.…”

Cytokinesis-block Micronucleus Assay by Manual and Automated Scoring

“…A promising strategy to increase throughput of cytogenetic biodosimetry achieving a rapid and reliable dose assessment for triage purposes in large scale nuclear or radiological accidents is the automation of scoring by sophisticated image analysis systems, and many works are in progress to validate automated CBMN analysis (Bolognesi et al 2011;Decordier et al 2011;Rossnerova et al 2011;Willems et al 2010).…”

“…The automation of scoring enables a faster delivery of results and reduces the variability between scorers. Several automated systems for the CBMN assay have been developed over the past two decades, but only in the past few years, more advanced systems have been available and undergone validation in studies of genotoxin exposure (Decordier et al 2011;Rossnerova et al 2011;Varga et al 2004) and of ionizing radiation exposure (Bolognesi et al 2011;Willems et al 2010). A quite recent automated system is Metafer4 (MetaSystems, Newton, MA, USA), consisting of a computer combined with a microscope fully motorized with a high resolution CCD camera and a motorized scanning stage (Schunck et al 2004).…”

HUMN project initiative and review of validation, quality control and prospects for further development of automated micronucleus assays using image cytometry systems

“…However, the experience of "omics" biomarkers, where reducing technical heterogeneity was a "standardise or die" condition, should be taken into consideration, and the identification of a set of minimal criteria, e.g., MIAME criteria for transcriptomics Varga et al (2004Varga et al ( , 2005Varga et al ( , 2006 Willems et al (2010) The correlation coefficient (R) between visual and automated scoring of MN in BN lymphocytes for base-line values was 0.84 and for ␥-irradiated samples at different doses it was 0.91; CV for repeat counts ranged from 5.4 to 9.3% depending on classifiers used . Subsequent studies by Willems et al (2010) Decordier et al (2009Decordier et al ( , 2011 Sensitivity: 75% of cells detected; >90% of MNs in BN and MN detected, i.e. the false negative rate is <10% specificity: the false positive MN rate is <1%.…”

“…The significant achievements, current state of application, restrictions and recommendations for future research for all the above-mentioned AIC CBMN assay systems were recently addressed in a special issue of Mutagenesis (Rossnerova et al, 2011a;Decordier et al, 2011;Darzynkiewicz et al, 2011). Advances in molecular cytogenetics (e.g.…”