Conventional, High-Resolution and Imaging Flow Cytometry: Benchmarking Performance in Characterisation of Extracellular Vesicles.
Conventional, High-Resolution and Imaging Flow Cytometry: Benchmarking Performance in Characterisation of Extracellular Vesicles.
复制标题
DOI：
10.3390/biomedicines9020124
复制
发表时间：
2021-01-27
期刊：
影响因子：
4.7
通讯作者：
Handberg A
中科院分区：
文献类型：
--
作者：
Botha J;Pugsley HR;Handberg A
流式细胞术仍然是一种常用的方法，因为它能够以高通量的方式表征单个颗粒上的多个参数。为了克服传统流式细胞术缺乏敏感性来表征细胞外小泡(EV)的局限性，人们开发了新的、高灵敏度的平台，如高分辨率和成像流式细胞仪。我们提供了常规FACS Aria III、高分辨率Apogee A60 Micro-plus和ImageStream X Mk II成像流式细胞仪平台的比较基准。纳米球被用来系统地表征每个平台检测和量化具有不同大小、折射率和荧光性质的种群的能力，并报告了每个种群在浓度测定中的重复性。我们评估了三个平台检测血浆中不同EV表型的能力，以及在确定EV浓度方面的日内、日间和全球变异性。通过应用这种或类似的方法来表征方法，研究人员将能够在知情的情况下做出选择平台的决定，从而能够根据每个项目的需求将合适的流式细胞仪平台与项目相匹配。这将大大有助于提高电动汽车研究的稳健性和重复性。
Flow cytometry remains a commonly used methodology due to its ability to characterise multiple parameters on single particles in a high-throughput manner. In order to address limitations with lacking sensitivity of conventional flow cytometry to characterise extracellular vesicles (EVs), novel, highly sensitive platforms, such as high-resolution and imaging flow cytometers, have been developed. We provided comparative benchmarks of a conventional FACS Aria III, a high-resolution Apogee A60 Micro-PLUS and the ImageStream X Mk II imaging flow cytometry platform. Nanospheres were used to systematically characterise the abilities of each platform to detect and quantify populations with different sizes, refractive indices and fluorescence properties, and the repeatability in concentration determinations was reported for each population. We evaluated the ability of the three platforms to detect different EV phenotypes in blood plasma and the intra-day, inter-day and global variabilities in determining EV concentrations. By applying this or similar methodology to characterise methods, researchers would be able to make informed decisions on choice of platforms and thereby be able to match suitable flow cytometry platforms with projects based on the needs of each individual project. This would greatly contribute to improving the robustness and reproducibility of EV studies.
登录
查看更多内容
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：
9.3
作者：
de Rond, Leonie;van der Pol, Edwin;Coumans, Frank A. W.
通讯作者：
Coumans, Frank A. W.
影响因子：
3.7
作者：
Erdbruegger, Uta;Rudy, Christine K.;Lannigan, Joanne
通讯作者：
Lannigan, Joanne
影响因子：
8.6
作者：
Knoner, Gregor;Parkin, Simon;Rubinsztein-Dunlop, Halina
通讯作者：
Rubinsztein-Dunlop, Halina
DOI：
10.1111/jth.12767
发表时间：
2015-02
期刊：
Journal of thrombosis and haemostasis : JTH
影响因子：
--
作者：
Arraud N;Gounou C;Linares R;Brisson AR
通讯作者：
Brisson AR