EuroFlow standardization of flow cytometer instrument settings and immunophenotyping protocols

被引：599

作者：

Kalina, T. ^{[2
]}

Flores-Montero, J. ^{[3
,4
,5
]}

van der Velden, V. H. J. ^{[1
]}

Martin-Ayuso, M. ^{[6
]}

Boettcher, S. ^{[7
]}

Ritgen, M. ^{[7
]}

Almeida, J. ^{[3
,4
,5
]}

Lhermitte, L. ^{[8
,9
]}

Asnafi, V. ^{[8
,9
]}

Mendonca, A. ^{[10
]}

de Tute, R. ^{[11
]}

Cullen, M. ^{[11
]}

Sedek, L. ^{[12
]}

Vidriales, M. B. ^{[13
,14
]}

Perez, J. J. ^{[13
,14
]}

te Marvelde, J. G. ^{[1
]}

Mejstrikova, E. ^{[2
]}

Hrusak, O. ^{[2
]}

Szczepanski, T. ^{[12
]}

van Dongen, J. J. M. ^{[1
]}

Orfao, A. ^{[3
,4
,5
]}

机构：

[1] Univ Med Ctr Rotterdam, Erasmus MC, Dept Immunol, NL-3015 GE Rotterdam, Netherlands

[2] Charles Univ DPH O, Fac Med 2, Dept Pediat Hematol & Oncol, Prague, Czech Republic

[3] Univ Salamanca USAL, Dept Med, Canc Res Ctr IBMCC CSIC, Salamanca, Spain

[4] Univ Salamanca USAL, Cytometry Serv, Salamanca, Spain

[5] Inst Biomed Res Salamanca IBSAL, Salamanca, Spain

[6] Cytognos SL, Salamanca, Spain

[7] Univ Hosp Schleswig Holstein, Dept Med 2, Kiel, Germany

[8] Univ Paris 05, Hop Necker, AP HP, Dept Hematol, Paris, France

[9] Univ Paris 05, AP HP, UMR CNRS 8147, Paris, France

[10] Portuguese Inst Oncol IPOLFG, Dept Hematol, Lisbon, Portugal

[11] Univ Leeds UNIVLEEDS, HMDS, Leeds, W Yorkshire, England

[12] Med Univ Silesia SUM, Dept Pediat Hematol & Oncol, Zabrze, Poland

[13] Univ Hosp Salamanca HUS, Dept Hematol, Salamanca, Spain

[14] IBSAL, Salamanca, Spain

来源：

LEUKEMIA | 2012年 / 26卷 / 09期

关键词：

flow cytometry; standardization; compensation; software; fluorochromes; immunophenotyping; MINIMAL RESIDUAL DISEASE; INTERNATIONAL CONSENSUS RECOMMENDATIONS; CHRONIC LYMPHOPROLIFERATIVE DISORDERS; ACUTE LYMPHOBLASTIC-LEUKEMIA; HEMATOLYMPHOID NEOPLASIA; HEMATOLOGIC NEOPLASIA; QUALITY-CONTROL; REAGENTS; VISUALIZATION; COMPENSATION;

D O I：

10.1038/leu.2012.122

中图分类号：

R73 [肿瘤学];

学科分类号：

100214 [肿瘤学];

摘要：

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.

引用

页码：1986 / 2010

页数：25

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