Comments on EuroFlow standard operating procedures for instrument setup and compensation for BD FACS Canto II, Navios and BD FACS Lyric instruments
Comments on EuroFlow standard operating procedures for instrument setup and compensation for BD FACS Canto II, Navios and BD FACS Lyric instruments
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DOI：
10.1016/j.jim.2019.112680
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发表时间：
2019-12-01
影响因子：
2.2
通讯作者：
Kalina, Tomas
中科院分区：
文献类型：
--
作者：
Glier, Hana;Novakova, Michaela;Kalina, Tomas
本评论讨论了EuroFlow标准化流式细胞仪分析的三种不同的流式细胞仪的应用的特殊性。EuroFlow联盟开发了一种完全标准化的方法，用于血液恶性肿瘤和原发性免疫缺陷的流式细胞术免疫表型分析。标准化仪器设置是EuroFlow标准化的重要组成部分。最初，EuroFlow Consortium开发并优化了逐步标准操作规程（SOP），以设置8色BD FACSCanto II流式细胞仪（Canto），后来纳入了Navios（Beckman Coulter）和BD FACSLyric（Lyric）。开发这些SOP是为了实现三种流式细胞仪中标准化和完全可比的荧光测量。在Canto和Navios中，使用Rainbow beads校准颗粒的参比峰的平均荧光强度（MFI）为各个仪器中的每个检测器通道设置光电倍增管（PMT）电压，以在不同仪器中达到相同的MFI。反过来，Lyric仪器的一个新功能允许共享用于以测定形式将阳性群体放置在仪器之间的相同位置的属性集合，作为其集成在细胞仪设置和跟踪（CS&T）模块中的组件之一。因此，EuroFlow Lyric分析允许在Lyric上标准化采集8色EuroFlow板，而无需手动设置单个仪器上的PMT电压。总之，EuroFlow开发的标准化仪器设置可实现流式细胞仪测量的跨平台实验室间和实验室内标准化。本评论提供了对Canto、Navios和Lyric仪器的标准化EuroFlow仪器设置进行修改的观点，这些修改在EuroFlow网站上提供的各个仪器专用SOP中进行了详细描述。
This commentary discusses particularities of application of the EuroFlow standardization of flow cytometric analyses on three different flow cytometers. The EuroFlow consortium developed a fully standardized approach for flow cytometric immunophenotyping of hematological malignancies and primary immunodeficiencies. Standardized instrument setup is an essential part of EuroFlow standardization. Initially, the EuroFlow Consortium developed and optimized a step-by-step standard operating procedure (SOP) to setup 8-color BD FACSCanto II flow cytometer (Canto), with the later inclusion of Navios (Beckman Coulter) and BD FACSLyric (Lyric). Those SOPs were developed to enable standardized and fully comparable fluorescence measurements in the three flow cytometers. In Canto and Navios, mean fluorescence intensity (MFI) of a reference peak of Rainbow beads calibration particles is used to set up photomultiplier (PMT) voltages for each detector channel in individual instruments to reach the same MFI across distinct instruments. In turn, a new feature of Lyric instruments allows to share collection of attributes that are used to place the positive population at the same position among instruments in the form of assays, as one of its components integrated in the Cytometer Setup and Tracking (CS&T) module. The EuroFlow Lyric assays thus allow for standardized acquisition of 8-color EuroFlow panels on Lyric without the need to setup the PMT voltages on the individual instruments manually.In summary, the standardized instrument setup developed by EuroFlow enables cross-platform inter- and infra-laboratory standardization of flow cytometric measurements. This commentary provides a perspective on the modifications of the standardized EuroFlow instrument setup of Canto, Navios and Lyric instruments that are described in detail in individual instrument-specfic SOPs available at the EuroFlow website.