title: Flow cytometry assay for counting micronucleated erythrocytes: development process.
abstract: Development of any new assay proceeds in several phases. When an assay is intended for regular use to support regulatory decision-making, there are significant additional stages in the development process beyond the initial description of the method. In this paper we discuss some of the studies related to the development of a flow cytometric method for counting micronuclei in rodent erythrocytes. Studies related to fixation methods and conditions, standardization of DNA staining, and antibody staining are discussed. These studies, while not part of the formal description of the method, are needed as part of the preparation for the formal validation of the method. In addition, the lessons learned in transferring the method to other laboratories are briefly discussed in relation to defining the final protocol.
authors.affiliation: Division of Applied Pharmacology Research, Food and Drug Administration, Laurel, Maryland, USA.
authors.forename: J L
authors.initials: JL
authors.lastname: Weaver
authors.forename: D
authors.initials: D
authors.lastname: Torous
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000032:analysis; D000944:Antigens, Differentiation, B-Lymphocyte / Q000032:analysis; D015415:Biomarkers; D004247:DNA / Q000032:analysis; D016054:DNA, Protozoan / Q000032:analysis; D004906:Erythrocyte Count / Q000379:methods; D004912:Erythrocytes / Q000469:parasitology / Q000473:pathology*; D005434:Flow Cytometry / Q000379:methods* / Q000592:standards; D005456:Fluorescent Dyes; D008288:Malaria / Q000097:blood / Q000469:parasitology / Q000473:pathology; D051379:Mice; D048631:Micronucleus, Germline / Q000469:parasitology / Q000473:pathology*; D010380:Peer Review; D010962:Plasmodium berghei / Q000235:genetics / Q000502:physiology; D051381:Rats; D011990:Receptors, Transferrin; D015203:Reproducibility of Results; D012156:Reticulocytes / Q000469:parasitology / Q000473:pathology; D013194:Staining and Labeling / Q000379:methods; D013696:Temperature; D016707:Tissue Fixation / Q000379:methods
chemical_list: D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; D015415:Biomarkers; C068126:CD71 antigen; D016054:DNA, Protozoan; D005456:Fluorescent Dyes; D011990:Receptors, Transferrin; D004247:DNA