title: EuroFlow standardization of flow cytometer instrument settings and immunophenotyping protocols.
abstract: The EU-supported EuroFlow Consortium aimed at innovation and standardization of immunophenotyping for diagnosis and classification of hematological malignancies by introducing 8-color flow cytometry with fully standardized laboratory procedures and antibody panels in order to achieve maximally comparable results among different laboratories. This required the selection of optimal combinations of compatible fluorochromes and the design and evaluation of adequate standard operating procedures (SOPs) for instrument setup, fluorescence compensation and sample preparation. Additionally, we developed software tools for the evaluation of individual antibody reagents and antibody panels. Each section describes what has been evaluated experimentally versus adopted based on existing data and experience. Multicentric evaluation demonstrated high levels of reproducibility based on strict implementation of the EuroFlow SOPs and antibody panels. Overall, the 6 years of extensive collaborative experiments and the analysis of hundreds of cell samples of patients and healthy controls in the EuroFlow centers have provided for the first time laboratory protocols and software tools for fully standardized 8-color flow cytometric immunophenotyping of normal and malignant leukocytes in bone marrow and blood; this has yielded highly comparable data sets, which can be integrated in a single database.
authors.affiliation: Department of Pediatric Hematology and Oncology, 2nd Faculty of Medicine, Charles University (DPH/O), Prague, Czech Republic.
authors.forename: T
authors.initials: T
authors.lastname: Kalina
authors.forename: J
authors.initials: J
authors.lastname: Flores-Montero
authors.forename: V H J
authors.initials: VH
authors.lastname: van der Velden
authors.forename: M
authors.initials: M
authors.lastname: Martin-Ayuso
authors.forename: S
authors.initials: S
authors.lastname: Böttcher
authors.forename: M
authors.initials: M
authors.lastname: Ritgen
authors.forename: J
authors.initials: J
authors.lastname: Almeida
authors.forename: L
authors.initials: L
authors.lastname: Lhermitte
authors.forename: V
authors.initials: V
authors.lastname: Asnafi
authors.forename: A
authors.initials: A
authors.lastname: Mendonça
authors.forename: R
authors.initials: R
authors.lastname: de Tute
authors.forename: M
authors.initials: M
authors.lastname: Cullen
authors.forename: L
authors.initials: L
authors.lastname: Sedek
authors.forename: M B
authors.initials: MB
authors.lastname: Vidriales
authors.forename: J J
authors.initials: JJ
authors.lastname: Pérez
authors.forename: J G
authors.initials: JG
authors.lastname: te Marvelde
authors.forename: E
authors.initials: E
authors.lastname: Mejstrikova
authors.forename: O
authors.initials: O
authors.lastname: Hrusak
authors.forename: T
authors.initials: T
authors.lastname: Szczepański
authors.forename: J J M
authors.initials: JJ
authors.lastname: van Dongen
authors.forename: A
authors.initials: A
authors.lastname: Orfao
mesh_terms: D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor / Q000276:immunology; D005060:Europe; D005434:Flow Cytometry / Q000295:instrumentation* / Q000379:methods / Q000592:standards*; D019337:Hematologic Neoplasms / Q000175:diagnosis* / Q000276:immunology*; D006801:Humans; D016130:Immunophenotyping / Q000592:standards*; D007753:Laboratories / Q000592:standards*; D011379:Prognosis
chemical_list: D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor