FCMPASS Software Aids Extracellular Vesicle Light Scatter Standardization
FCMPASS Software Aids Extracellular Vesicle Light Scatter Standardization
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DOI：
10.1002/cyto.a.23782
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发表时间：
2019-06-28
期刊：
影响因子：
3.7
通讯作者：
Englyst, Nicola A.
中科院分区：
文献类型：
--
作者：
Welsh, Joshua A.;Horak, Peter;Englyst, Nicola A.
细胞外囊泡（EV）的研究是一个快速增长的领域，由于其在临床医学的许多领域，包括诊断学，诊断学，治疗诊断学和治疗学的巨大潜力。流式细胞术是目前最流行的分析EV的方法之一，因为它是一种高通量、多参数的技术，在大多数研究实验室中都很容易获得。尽管其广泛使用，但很少有商业流式细胞仪专门设计用于检测EV。许多用于EV分析的流式细胞仪在其检测限下工作，无法检测大多数EV。目前，EV流式细胞术几乎没有标准化，这是一个问题，因为流式细胞仪在收集来自被询问颗粒的散射光或荧光的方式上变化很大。这使得已发表的研究很难解释、比较，在某些情况下，甚至不可能复制。在这里，我们展示了流式细胞仪光散射标准化的方法，利用流式细胞仪采集后分析软件（FCMPASS）。FCMPASS建立在Mie理论基础上，通过分析已知直径和折射率的微珠或通过输入收集角（如果已知），可以近似流式细胞仪的几何参数。然后，该软件能够创建散射直径曲线和散射折射率曲线，使研究人员能够将散射数据和仪器灵敏度转换为标准化单位。此外，通过正确的控制，光散射数据可以转换为直径分布或折射率分布。因此，FCMPASS提供了一种免费提供的符合人体工程学的标准化方法，并使用流式细胞术进一步扩展EV表征。(c)2019作者Cytometry Part A由Wiley Periodicals，Inc.出版。代表国际细胞计数促进协会。
The study of extracellular vesicles (EVs) is a rapidly growing field due to their great potential in many areas of clinical medicine including diagnostics, prognostics, theranostics, and therapeutics. Flow cytometry is currently one of the most popular methods of analyzing EVs due to it being a high-throughput, multiparametric technique, that is readily available in the majority of research labs. Despite its wide use, few commercial flow cytometers are designed specifically for the detection of EVs. Many flow cytometers used for EV analysis are working at their detection limits and are unable to detect the majority of EVs. Currently, very little standardization exists for EV flow cytometry, which is an issue because flow cytometers vary considerably in the way they collect scattered or fluorescent light from particles being interrogated. This makes published research hard to interpret, compare, and in some cases, impossible to reproduce. Here we demonstrate a method of flow cytometer light scatter standardization, utilizing flow cytometer postacquisition analysis software (FCMPASS). FCMPASS is built upon Mie theory and enables the approximation of flow cytometer geometric parameters either by analyzing beads of known diameter and refractive index or by inputting the collection angle if known. The software is then able to create a scatter-diameter curve and scatter-refractive index curve that enables researchers to convert scattering data and instrument sensitivity into standardized units. Furthermore, with the correct controls, light scatter data can be converted to diameter distributions or refractive index distributions. FCMPASS therefore offers a freely available and ergonomic method of standardizing and further extending EV characterization using flow cytometry. (c) 2019 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of International Society for Advancement of Cytometry.