EuroFlow standardization of flow cytometer instrument settings and immunophenotyping protocols

Autor: J. G. te Marvelde, Lukasz Sedek, Alberto Orfao, Ester Mejstrikova, M Cullen, V H J van der Velden, T Szczepa nacute, Ondrej Hrusak, S Böttcher, R de Tute, ski, Matthias Ritgen, Marta Martin-Ayuso, Vahid Asnafi, Alexandre de Mendonça, J.J. Pérez, Juan Flores-Montero, M B Vidriales, J. J. M. Van Dongen, Ludovic Lhermitte, Julia Almeida, Tomas Kalina
Přispěvatelé: Immunology, Junta de Castilla y León, Instituto de Salud Carlos III, Ministry of Education, Youth and Sports (Czech Republic), European Commission, Ministry of Health of the Czech Republic, Red Temática de Investigación Cooperativa en Cáncer (España), Ministerio de Ciencia e Innovación (España), Cytokines, hématopoïèse et réponse immune (CHRI), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université Paris Descartes - Paris 5 (UPD5) - Institut National de la Santé et de la Recherche Médicale (INSERM) - Centre National de la Recherche Scientifique (CNRS)
Rok vydání: 2012
Předmět:
Zdroj: Leukemia
Leukemia, 26(9), 1986-2010. Nature Publishing Group
Leukemia, Nature Publishing Group: Open Access Hybrid Model Option B, 2012, 26, pp.1886-2010
Digital.CSIC. Repositorio Institucional del CSIC
instname
ISSN: 1476-5551
0887-6924
Popis: This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivative Works 3.0 Unported License.-- EuroFlow Consortium (EU-FP6, LSHB-CT-2006-018708): et al.
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.
The research activities of the EuroFlow Consortium were supported by the European Commission (grant STREP EU-FP6, LSHB-CT-2006-018708, entitled ‘Flow cytometry for fast and sensitive diagnosis and follow-up of hematological malignancies’) and the following national grants: Spanish Network of Cancer Research Centers (ISCIII RTICC-RD06/0020/0035-FEDER), FIS 08/90881 from the ‘Fondo de Investigación Sanitaria’, Ministerio de Ciencia e Innovación (Madrid, Spain), SA016-A-09 from the Consejería de Educación, Junta de Castilla y León, Valladolid, Spain, and PIB2010BZ-00565 from the Dirección General de Cooperación Internacional y Relaciones Institucionales, Secretaría de Estado de Investigación, Ministerio de Ciencia e Innovación (Madrid, Spain). T Kalina, E Mejstrikova and O Hrusak were supported by the Czech Ministry of Education Grant No. MSM0021620813, Czech Ministry of Health grant NT/12425-4 and TK is supported as an ISAC Scholar by The International Society for Advancement of Cytometry.
Databáze: OpenAIRE
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