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Flow cytometric enumeration of micronucleated reticulocytes: High transferability among 14 laboratories
Author(s) -
Torous Dorothea K.,
Hall Nikki E.,
Dertinger Stephen D.,
Diehl Marilyn S.,
IlliLove Anne H.,
Cederbrant Karin,
Sandelin Kerstin,
Bolcsfoldi George,
Ferguson Lynnette R.,
Pearson Amira,
Majeska Jenness B.,
Tarca James P.,
Hewish Dean R.,
Doughty Larissa,
Fenech Michael,
Weaver James L.,
Broud Dennis D.,
Gatehouse David G.,
Hynes Geoffrey M.,
Kwanyuen Puntipa,
McLean Jack,
McNamee James P.,
Parenteau Monique,
Van Hoof Veerle,
Vanparys Philippe,
Lenarczyk Marek,
Siennicka Joanna,
Litwinska Bogumila,
Slowikowska Maria G.,
Harbach Philip R.,
Johnson Carol W.,
Zhao Shuou,
Aaron Charles S.,
Lynch Anthony M.,
Marshall Ian C.,
Rodgers Brenda,
Tometsko Carol R.
Publication year - 2001
Publication title -
environmental and molecular mutagenesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1
H-Index - 87
eISSN - 1098-2280
pISSN - 0893-6692
DOI - 10.1002/em.1051
Subject(s) - plasmodium berghei , micronucleus test , biology , reticulocyte , malaria , transferability , enumeration , immunology , genetics , chemistry , gene , statistics , mathematics , organic chemistry , logit , toxicity , combinatorics , messenger rna
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. Environ. Mol. Mutagen. 38:59–68, 2001 © 2001 Wiley‐Liss, Inc.