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ToxSci Advance Access published online on May 28, 2003
Toxicological Sciences, doi:10.1093/toxsci/kfg143
Toxicological Sciences © Society of Toxicology 2003; all rights reserved
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Received March 31, 2003; accepted April 30, 2003
© 2003 Society of Toxicology

Genetic Toxicology

Comparative Scoring of Micronucleated Reticulocytes in Rat Peripheral Blood by Flow Cytometry and Microscopy

Dorothea K. Torous 1, Nikki E. Hall 1, Francis G. Murante 1, Sarah E. Gleason 1, Carol R. Tometsko 1, Stephen D. Dertinger 1*

1 Litron Laboratories, 1351 Mount Hope Ave., Rochester, NY

* To whom correspondence should be addressed. E-mail: sdertinger{at}litronlabs.com.


  Abstract

A flow cytometric technique for scoring the incidence of micronucleated reticulocytes in rat peripheral blood was compared to a standard microscopy-based procedure. For these studies, groups of five male Sprague-Dawley rats were treated with vehicle or a broad range of chemical genotoxicants: 6-thioguanine, N-methyl-N'-nitro-N-nitrosoguanidine, vincristine, methylaziridine, acetaldehyde, methyl methanesulfonate, benzene, monocrotaline, and azathioprine. Animals were treated once a day for up to two days, and peripheral blood was collected between twenty-four and forty-eight hours after the final administration. These samples were processed for flow cytometric scoring and microscopy-based analysis using supravital acridine orange staining, and the percentage of reticulocytes and micronucleated reticulocytes was determined for each sample. The resulting data demonstrate good agreement between these scoring methodologies, although careful execution of the flow cytometric method was found to enhance the micronucleus assay by reducing both scoring time and scoring error. These data add further support to the premise that the peripheral blood compartment of rats can be used effectively to detect genotoxicant-induced micronuclei.

Key Words: Micronuclei, Genotoxicity, Flow cytometry, Rats, CD71 antigen .


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