EuroFlow standardization of flow cytometer instrument settings and immunophenotyping protocols

• Published in: Leukemia Vol. 26; no. 9; pp. 1986 - 2010
• Main Authors: Kalina, T, Flores-Montero, J, van der Velden, V H J, Martin-Ayuso, M, Böttcher, S, Ritgen, M, Almeida, J, Lhermitte, L, Asnafi, V, Mendonça, A, de Tute, R, Cullen, M, Sedek, L, Vidriales, M B, Pérez, J J, te Marvelde, J G, Mejstrikova, E, Hrusak, O, Szczepański, T, van Dongen, J J M, Orfao, A
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.09.2012; Nature Publishing Group; Springer Nature
• Subjects: 631/114/794; 631/1647/1407/1492; 692/699/67/1990; Antibodies; Antibodies, Monoclonal; Biomarkers, Tumor - immunology; Bone marrow; Cancer Research; Consortia; Critical Care Medicine; Data analysis; Diagnosis; Europe; Flow cytometry; Flow Cytometry - instrumentation; Flow Cytometry - methods; Flow Cytometry - standards; Hematologic Neoplasms - diagnosis; Hematologic Neoplasms - immunology; Hematology; Human health and pathology; Humans; Immunology; Immunophenotyping - standards; Intensive; Internal Medicine; Laboratories; Laboratories - standards; Leukemia; Life Sciences; Medicine; Medicine & Public Health; Oncology; Pediatrics; Physiological aspects; Prognosis; Reagents; Reproducibility; Software utilities; Special Report; Standardization; Viral antibodies; Europe; Czech Republic; Netherlands; Germany; Spain; standardization; flow cytometry; compensation; software; immunophenotyping; fluorochromes
• ISSN: 0887-6924; 1476-5551
• DOI: 10.1038/leu.2012.122

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.