title: Laser scanning cytometry for automation of the micronucleus assay.
abstract: Laser scanning cytometry (LSC) provides a novel approach for automated scoring of micronuclei (MN) in different types of mammalian cells, serving as a biomarker of genotoxicity and mutagenicity. In this review, we discuss the advances to date in measuring MN in cell lines, buccal cells and erythrocytes, describe the advantages and outline potential challenges of this distinctive approach of analysis of nuclear anomalies. The use of multiple laser wavelengths in LSC and the high dynamic range of fluorescence and absorption detection allow simultaneous measurement of multiple cellular and nuclear features such as cytoplasmic area, nuclear area, DNA content and density of nuclei and MN, protein content and density of cytoplasm as well as other features using molecular probes. This high-content analysis approach allows the cells of interest to be identified (e.g. binucleated cells in cytokinesis-blocked cultures) and MN scored specifically in them. MN assays in cell lines (e.g. the CHO cell MN assay) using LSC are increasingly used in routine toxicology screening. More high-content MN assays and the expansion of MN analysis by LSC to other models (i.e. exfoliated cells, dermal cell models, etc.) hold great promise for robust and exciting developments in MN assay automation as a high-content high-throughput analysis procedure.
authors.affiliation: Department of Pathology, New York Medical College, Valhalla, NY 10595, USA. darzynk@nymc.edu
authors.forename: Zbigniew
authors.initials: Z
authors.lastname: Darzynkiewicz
authors.forename: Piotr
authors.initials: P
authors.lastname: Smolewski
authors.forename: Elena
authors.initials: E
authors.lastname: Holden
authors.forename: Ed
authors.initials: E
authors.lastname: Luther
authors.forename: Mel
authors.initials: M
authors.lastname: Henriksen
authors.forename: Maxime
authors.initials: M
authors.lastname: François
authors.forename: Wayne
authors.initials: W
authors.lastname: Leifert
authors.forename: Michael
authors.initials: M
authors.lastname: Fenech
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000032:analysis; D004912:Erythrocytes / Q000648:ultrastructure; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D047569:Laser Scanning Cytometry / Q000379:methods*; D015162:Micronucleus Tests; D009061:Mouth Mucosa / Q000648:ultrastructure
chemical_list: D004247:DNA