Abstract：
Nanoparticle tracking analysis (NTA) has been one of several characterization methods used for extracellular vesicle (EV) research since 2006. Many consider that NTA instruments and their software packages can be easily utilized following minimal training and that size calibration is feasible in-house. As both NTA acquisition and software analysis constitute EV characterization, they are addressed in Minimal Information for Studies of Extracellular Vesicles 2018 (MISEV2018). In addition, they have been monitored by Transparent Reporting and Centralizing Knowledge in Extracellular Vesicle Research (EV-TRACK) to improve the robustness of EV experiments (e.g., minimize experimental variation due to uncontrolled factors). Despite efforts to encourage the reporting of methods and controls, many published research papers fail to report critical settings needed to reproduce the original NTA observations. Few papers report the NTA characterization of negative controls or diluents, evidently assuming that commercially available products, such as phosphate-buffered saline or ultrapure distilled water, are particulate-free. Similarly, positive controls or size standards are seldom reported by researchers to verify particle sizing. The Stokes-Einstein equation incorporates sample viscosity and temperature variables to determine particle displacement. Reporting the stable laser chamber temperature during the entire sample video collection is, therefore, an essential control measure for accurate replication. The filtration of samples or diluents is also not routinely reported, and if so, the specifics of the filter (manufacturer, membrane material, pore size) and storage conditions are seldom included. The International Society for Extracellular Vesicle (ISEV)\'s minimal standards of acceptable experimental detail should include a well-documented NTA protocol for the characterization of EVs. The following experiment provides evidence that an NTA analysis protocol needs to be established by the individual researcher and included in the methods of publications that use NTA characterization as one of the options to fulfill MISEV2018 requirements for single vesicle characterization.
摘要：
自2006年以来，纳米颗粒跟踪分析（NTA）一直是用于细胞外囊泡（EV）研究的几种表征方法之一。许多人认为，经过最少的培训后，NTA仪器及其软件包可以轻松使用，并且内部尺寸校准是可行的。由于NTA采集和软件分析都构成了EV表征，它们在2018年细胞外囊泡研究的最小信息（MISEV2018）中得到了解决。此外,它们已通过透明报告和细胞外囊泡研究集中知识(EV-TRACK)进行监测，以提高EV实验的稳健性(例如，最小化由于不受控制的因素导致的实验变化)。尽管努力鼓励报告方法和控制措施，许多已发表的研究论文未能报告重现原始NTA观察结果所需的关键设置。很少有论文报道阴性对照或稀释剂的NTA表征，显然假设市售产品，如磷酸盐缓冲盐水或超纯蒸馏水，没有微粒。同样，研究人员很少报告阳性对照或大小标准来验证颗粒大小。斯托克斯-爱因斯坦方程结合了样品粘度和温度变量来确定颗粒位移。在整个样本视频采集期间报告稳定的激光室温度，因此,精确复制的重要控制措施。样品或稀释剂的过滤也没有常规报道，如果是这样,过滤器的细节(制造商，膜材料，孔径）和储存条件很少包括在内。国际细胞外囊泡学会（ISEV）的可接受实验细节的最低标准应包括一个有据可查的NTA方案，用于表征电动汽车。以下实验提供了证据，表明需要由个体研究人员建立NTA分析方案，并将其包括在使用NTA表征作为满足MISEV2018单囊泡表征要求的选项之一的出版物方法中。
