Sizing and phenotyping of cellular vesicles using Nanoparticle Tracking Analysis.
Sizing and phenotyping of cellular vesicles using Nanoparticle Tracking Analysis.
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DOI：
10.1016/j.nano.2011.04.003
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发表时间：
2011-12
影响因子：
5.4
通讯作者：
Sargent, Ian L.
中科院分区：
文献类型：
--
作者：
Dragovic, Rebecca A.;Gardiner, Christopher;Brooks, Alexandra S.;Tannetta, Dionne S.;Ferguson, David J. P.;Hole, Patrick;Carr, Bob;Redman, Christopher W. G.;Harris, Adrian L.;Dobson, Peter J.;Harrison, Paul;Sargent, Ian L.
细胞微泡和纳米泡（外泌体）参与许多疾病过程，并且具有作为生物标志物的巨大潜力。然而，该领域的发展受到可用于其测量的技术的限制。在这里，我们报告了使用荧光纳米颗粒跟踪分析（NTA）来快速确定细胞囊泡的大小和表型。在该系统中，使用光学显微镜通过光散射来观察囊泡。拍摄视频后，NTA 软件会跟踪单个囊泡的布朗运动并计算它们的大小和总浓度。使用人胎盘囊泡和血浆，我们证明 NTA 可以测量小至~50 nm 的细胞囊泡，并且比传统流式细胞术（下限~300 nm）灵敏得多。通过将 NTA 与荧光测量相结合，我们证明了可以用特定的抗体偶联量子点标记囊泡，从而确定其表型。这项研究的作者利用荧光纳米颗粒跟踪分析 (NTA) 来快速确定细胞囊泡的大小和表型，证明 NTA 比传统流式细胞术灵敏得多。胎盘囊泡光散射视频的屏幕截图，上面覆盖了通过纳米颗粒跟踪分析确定的囊泡大小和浓度图。
Cellular microvesicles and nanovesicles (exosomes) are involved in many disease processes and have major potential as biomarkers. However, developments in this area are constrained by limitations in the technology available for their measurement. Here we report on the use of fluorescence nanoparticle tracking analysis (NTA) to rapidly size and phenotype cellular vesicles. In this system vesicles are visualized by light scattering using a light microscope. A video is taken, and the NTA software tracks the brownian motion of individual vesicles and calculates their size and total concentration. Using human placental vesicles and plasma, we have demonstrated that NTA can measure cellular vesicles as small as ∼50 nm and is far more sensitive than conventional flow cytometry (lower limit ∼300 nm). By combining NTA with fluorescence measurement we have demonstrated that vesicles can be labeled with specific antibody-conjugated quantum dots, allowing their phenotype to be determined. The authors of this study utilized fluorescence nanoparticle tracking analysis (NTA) to rapidly size and phenotype cellular vesicles, demonstrating that NTA is far more sensitive than conventional flow cytometry. Screen shot from video of light scatter of placental vesicles overlaid with a graph of vesicle size and concentration as determined by nanoparticle tracking analysis.
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影响因子：
3.8
作者：
Redman, C. W. G.;Sargent, I. L.
通讯作者：
Sargent, I. L.
影响因子：
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Umezawa, N;Gelman, MA;Gellman, SH
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Gellman, SH
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Harrison, P.
影响因子：
3.7
作者：
Orozco, Aaron F.;Lewis, Dorothy E.
通讯作者：
Lewis, Dorothy E.