Camacho Virginia, Toxavidis Vasilis, Tigges John C
Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA.
Flow Cytometry Core, Beth Israel Deaconess Medical Center, CLS-0932, 330 Brookline Ave., Boston, MA, 02215, USA.
Methods Mol Biol. 2017;1660:175-190. doi: 10.1007/978-1-4939-7253-1_14.
Here, we describe a comprehensive methodology for the setup and standardization of EV analysis using nanoscale flow cytometry. Controls of different size ranges, fluorescent intensities, and materials can be used to set up distribution curves that are then used for instrument optimization and as a reference guide. Using these controls, flow cytometry instruments can be primed for the detection, analysis, and sorting of specific EV populations. This allows for cross platform comparison and the ability to monitor both quality control (QC) and quality assurance (QA). The method here describes the use of nanoparticles to optimize a flow cytometer for small particle detection. It also outlines the procedures necessary to recover EVs for downstream applications.
在此，我们描述了一种使用纳米级流式细胞术进行细胞外囊泡（EV）分析的设置和标准化的综合方法。不同尺寸范围、荧光强度和材料的对照可用于建立分布曲线，然后将其用于仪器优化并作为参考指南。使用这些对照，流式细胞仪可做好检测、分析和分选特定EV群体的准备。这允许进行跨平台比较，并具备监测质量控制（QC）和质量保证（QA）的能力。此处所述方法描述了使用纳米颗粒来优化流式细胞仪以进行小颗粒检测。它还概述了为下游应用回收EV所需的程序。
