PMID: 9186575May 23, 1997Paper

Flow cytometric analysis of micronucleated reticulocytes in mouse bone marrow

Mutation Research
S D DertingerK R Tometsko

Abstract

This laboratory has previously reported a flow cytometric procedure for quantitatively analyzing mouse peripheral blood reticulocytes for micronucleus content. The current study extends this line of investigation by evaluating whether these same flow cytometric scoring procedures can be applied to the analysis of mouse bone marrow samples. To validate the method, three groups of male BALB/c mice were treated with 100 mg/kg b.wt. methyl methanesulfonate. Bone marrow samples were collected 20, 40 or 60 h after administration. A set of 5 untreated animals was included to provide an indication of spontaneous micronucleus frequencies. The cells were fixed with ultracold methanol, treated with ribonuclease, and labeled with anti-CD71 antibody (FITC conjugate) and propidium iodide. This fixing and labeling procedure resulted in the resolution of the micronucleated reticulocyte population and facilitated high-speed acquisition and enumeration via flow cytometry. The number of micronucleated reticulocytes was determined flow cytometrically by the analysis of 10,000 total reticulocytes per bone marrow sample. In addition to these automated measurements, slides stained with acridine orange were prepared and the number of micronuclei per 1...Continue Reading

Citations

Jul 24, 2007·Nature Reviews. Drug Discovery·Jeffrey A KramerDale L Morris
May 16, 2007·Proceedings of the National Academy of Sciences of the United States of America·J ShugaL G Griffith
Jul 27, 2001·Environmental and Molecular Mutagenesis·D K TorousC R Tometsko

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