Flow cytometric enumeration of micronucleated reticulocytes: High transferability among 14 laboratories

Dorothea K. Torous, Nikki E. Hall, Stephen D. Dertinger, Marilyn S. Diehl, Anne H. Illi-Love, Karin Cederbrant, Kerstin Sandelin, George Bolcsfoldi, Lynnette R. Ferguson, Amira Pearson, Jenness B. Majeska, James P. Tarca, Dean R. Hewish, Larissa Doughty, Michael Fenech, James L. Weaver, Dennis D. Broud, David G. Gatehouse, Geoffrey M. Hynes, Puntipa KwanyuenJack McLean, James P. McNamee, Monique Parenteau, Veerle Van Hoof, Philippe Vanparys, Marek Lenarczyk, Joanna Siennicka, Bogumila Litwinska, Maria G. Slowikowska, Philip R. Harbach, Carol W. Johnson, Shuou Zhao, Charles S. Aaron, Anthony M. Lynch, Ian C. Marshall, Brenda Rodgers, Carol R. Tometsko

Research output: Contribution to journalArticlepeer-review

47 Scopus citations

Abstract

This laboratory previously described a single-laser flow cytometric method, which effectively resolves micronucleated erythrocyte populations in rodent peripheral blood samples. Even so, the rarity and variable size of micronuclei make it difficult to configure instrument settings consistently and define analysis regions rationally to enumerate the cell populations of interest. Murine erythrocytes from animals infected with the malaria parasite Plasmodium berghei contain a high prevalence of erythrocytes with a uniform DNA content. This biological model for micronucleated erythrocytes offers a means by which the micronucleus analysis regions can be rationally defined, and a means for controlling interexperimental variation. The experiments described herein were performed to extend these studies by testing whether malaria-infected erythrocytes could also be used to enhance the transferability of the method, as well as control intra- and interlaboratory variation. For these studies, blood samples from mice infected with malaria, or treated with vehicle or the clastogen methyl methanesulfonate, were fixed and shipped to collaborating laboratories for analysis. After configuring instrumentation parameters and guiding the position of analysis regions with the malaria-infected blood samples, micronucleated reticulocyte frequencies were measured (20,000 reticulocytes per sample). To evaluate both intra- and interlaboratory variation, five replicates were analyzed per day, and these analyses were repeated on up to five separate days. The data of 14 laboratories presented herein indicate that transferability of this flow cytometric technique is high when instrumentation is guided by the biological standard Plasmodium berghei.

Original languageEnglish
Pages (from-to)59-68
Number of pages10
JournalEnvironmental and Molecular Mutagenesis
Volume38
Issue number1
DOIs
StatePublished - 2001

Keywords

  • Flow cytometry
  • Genotoxicity
  • Interlaboratory
  • Micronucleus test
  • Validation

Fingerprint Dive into the research topics of 'Flow cytometric enumeration of micronucleated reticulocytes: High transferability among 14 laboratories'. Together they form a unique fingerprint.

Cite this