EuroFlow standardization of flow cytometer instrument settings and immunophenotyping protocols.

EuroFlow standardization of flow cytometer instrument settings and immunophenotyping protocols.
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DOI:
10.1038/leu.2012.122
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发表时间:
2012-09
期刊:
影响因子:
11.4
通讯作者:
Orfao, A.
Orfao, A.
中科院分区:
医学1区
文献类型:
--
作者:
Kalina, T.;Flores-Montero, J.;van der Velden, V. H. J.;Martin-Ayuso, M.;Boettcher, S.;Ritgen, M.;Almeida, J.;Lhermitte, L.;Asnafi, V.;Mendonca, A.;de Tute, R.;Cullen, M.;Sedek, L.;Vidriales, M. B.;Perez, J. J.;te Marvelde, J. G.;Mejstrikova, E.;Hrusak, O.;Szczepanski, T.;van Dongen, J. J. M.;Orfao, A.

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欧盟支持的EuroFlow联盟旨在通过引入具有完全标准化的实验室程序和抗体面板的8色流式细胞术来创新和标准化用于血液恶性肿瘤诊断和分类的免疫表型,以便在不同实验室之间实现最大限度的可比结果。这需要选择兼容荧光染料的最佳组合,并为仪器设置、荧光补偿和样品制备设计和评估适当的标准操作程序(SOP)。此外,我们还开发了用于评估单个抗体试剂和抗体小组的软件工具。每一节都描述了根据现有数据和经验进行的实验评估和采用的评估。多中心评估表明,在严格执行EuroFlow SOP和抗体小组的基础上,具有很高的重复性。总体而言,EuroFlow中心6年来的广泛合作实验和对患者和健康对照数百个细胞样本的分析首次为完全标准化的8色流式细胞仪免疫表型分析骨髓和血液中的正常和恶性白细胞提供了实验室方案和软件工具;这产生了高度可比的数据集,可以整合到一个数据库中。
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.
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