Open AccessAutomation of mouse micronucleus genotoxicity assay by laser scanning cytometry
Author(s) -
Styles J.A.,
Clark H.,
Festing M.F.W.,
Rew D.A.
Publication year - 2001
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/1097-0320(20010601)44:2<153::aid-cyto1095>3.0.co;2-h
Subject(s) - genotoxicity , micronucleus test , acridine orange , cytometry , micronucleus , propidium iodide , flow cytometry , microbiology and biotechnology , biology , chemistry , toxicity , biochemistry , apoptosis , organic chemistry , programmed cell death
Background The evaluation of the safety of drugs and other chemicals is an important aspect of toxicology work. The mouse micronucleus assay is a standard in vivo genotoxicity assay. Chromosomal damage is an indicator of genotoxicity, which manifests in the formation of micronuclei in polychromatic erythrocytes from bone marrow and in peripheral blood erythrocytes. The assay is laborious to perform by manual counting. The laser scanning cytometer allows automated and rapid quantitation of cellular and subcellular fluorescence in monodisperse cell samples on a microscope slide. The object of this study was to evaluate the application of this new technology in the mouse micronucleus genotoxicity assay. Materials and Methods One hundred forty‐four mice of various strains were dosed with combinations of carcinogens and antioxidants. Duplicate blood films were prepared 3 days later. One set of slides was stained with acridine orange, and the proportion of micronucleated erythrocytes was counted in 5,000 cells per slide. The duplicates were stained with propidium iodide (40 μg/ml). Five thousand cells per sample were examined using a laser scanning cytometer. The proportion of micronucleated erythrocytes was measured. Results A coefficient of correlation of 0.96 was found between the data from the two assays. The automation of the assay on the LSC produced a considerable time saving and efficiency gain. Conclusions We conclude that with further development, laser scanning cytometry is likely to become the preferred modality for the performance of standard genotoxicity assays. Cytometry 44:153–155, 2001. © 2001 Wiley‐Liss, Inc.