Improving Reproducibility to Meet Minimal Information for Studies of Extracellular Vesicles 2018 Guidelines in Nanoparticle Tracking Analysis.
Improving Reproducibility to Meet Minimal Information for Studies of Extracellular Vesicles 2018 Guidelines in Nanoparticle Tracking Analysis.
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DOI：
10.3791/63059
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发表时间：
2021-11-17
期刊：
影响因子：
--
通讯作者：
Weiss ML
中科院分区：
文献类型：
--
作者：
Snyder OL;Campbell AW;Christenson LK;Weiss ML
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，NTA仪器及其软件包可以很容易地使用后，最低限度的培训和大小校准是可行的内部。由于NTA采集和软件分析均构成EV表征，因此在细胞外囊泡研究的最小信息2018（MISEV 2018）中进行了讨论。此外，它们还受到细胞外囊泡研究中的透明报告和集中知识（EV-TRACK）的监测，以提高EV实验的稳健性（例如，最小化由于不可控因素引起的实验变化）。尽管努力鼓励报告的方法和控制，许多已发表的研究论文未能报告的关键设置需要复制原始NTA的意见。很少有论文报告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）不含微粒。同样，研究人员很少报告阳性对照或尺寸标准来验证颗粒尺寸。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。因此，在整个样品视频采集期间报告稳定的激光腔室温度是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告，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）可接受的实验细节的最低标准应包括一个文件齐全的NTA协议，用于EV的表征。以下实验提供的证据表明，NTA分析方案需要由单个研究人员建立，并包含在使用NTA表征的出版物方法中，作为满足MISEV 2018对单个囊泡表征要求的选项之一。
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.
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影响因子：
16
作者：
Lötvall J;Hill AF;Hochberg F;Buzás EI;Di Vizio D;Gardiner C;Gho YS;Kurochkin IV;Mathivanan S;Quesenberry P;Sahoo S;Tahara H;Wauben MH;Witwer KW;Théry C
通讯作者：
Théry C
影响因子：
3.1
作者：
Gollwitzer, Christian;Bartczak, Dorota;Varga, Zoltan
通讯作者：
Varga, Zoltan
影响因子：
10.4
作者：
van der Pol, E.;Coumans, F. A. W.;Nieuwland, R.
通讯作者：
Nieuwland, R.
DOI：
10.1016/j.jconrel.2014.12.041
发表时间：
2015-02-28
期刊：
Journal of controlled release : official journal of the Controlled Release Society
影响因子：
--
作者：
Maas SL;de Vrij J;van der Vlist EJ;Geragousian B;van Bloois L;Mastrobattista E;Schiffelers RM;Wauben MH;Broekman ML;Nolte-'t Hoen EN
通讯作者：
Nolte-'t Hoen EN
影响因子：
5.4
作者：
Danaei M;Dehghankhold M;Ataei S;Hasanzadeh Davarani F;Javanmard R;Dokhani A;Khorasani S;Mozafari MR
通讯作者：
Mozafari MR