Department of Pediatric Hematology and Oncology, 2nd Faculty of Medicine, Charles University (DPH/O), Prague, Czech Republic.
Leukemia. 2012 Sep;26(9):1986-2010. 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.
欧盟支持的 EuroFlow 联盟旨在通过引入 8 色流式细胞术和完全标准化的实验室程序和抗体面板，实现不同实验室之间的最大可比性，从而实现血液恶性肿瘤的诊断和分类的免疫表型创新和标准化。这需要选择最佳的兼容荧光染料组合，并设计和评估适当的标准操作程序 (SOP)，以进行仪器设置、荧光补偿和样品制备。此外，我们还开发了用于评估单个抗体试剂和抗体面板的软件工具。每个部分都描述了根据现有数据和经验，通过实验评估和采用的内容。多中心评估表明，基于严格执行 EuroFlow SOP 和抗体面板，具有高度的可重复性。总的来说，6 年来广泛的协作实验以及在 EuroFlow 中心对数百个患者和健康对照者的细胞样本进行的分析，首次为骨髓和血液中正常和恶性白细胞的完全标准化 8 色流式细胞术免疫表型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，可以集成到一个单一的数据库中。
