Label-free imaging and biomarker analysis of exosomes with plasmonic scattering microscopy.

Label-free imaging and biomarker analysis of exosomes with plasmonic scattering microscopy.
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DOI:
10.1039/d2sc05191e
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发表时间:
2022-11-09
期刊:
影响因子:
8.4
通讯作者:
Wang, Shaopeng
Wang, Shaopeng
中科院分区:
化学1区
文献类型:
--
作者:
Zhang, Pengfei;Jiang, Jiapei;Zhou, Xinyu;Kolay, Jayeeta;Wang, Rui;Wan, Zijian;Wang, Shaopeng

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外泌体分析是临床和生物学研究应用的有前途的工具。然而,外来体的检测和生物标志物定量在技术上具有挑战性,因为它们很小并且高度异质。在这里,我们报告了一种光学方法,用于成像外来体和定量其蛋白质标记物,而无需使用等离子体散射显微镜(PSM)的标签。与传统的表面等离子体共振(SPR)显微镜相比,PSM可以提供更高的空间分辨率和无失真的图像,具有与物镜耦合表面等离子体共振(SPR)显微镜相似的信噪比,以及作为棱镜耦合SPR系统的毫米级视场,从而允许高通量的外泌体尺寸分布分析。此外,PSM保留了SPR传感器的高特异性和表面灵敏度,因此允许从具有抗体修饰的传感器表面的细胞外囊泡中选择外泌体,并原位分析抗体与捕获的外泌体上的表面蛋白质生物标志物之间的结合动力学。最后,PSM可以很容易地构建在一个流行的棱镜耦合SPR系统与商业可用的组件。因此,它可以为临床外泌体分析和探索诸如外泌体生物标志物结合特性的基本问题提供经济且强大的工具。散射成像方案允许常规的基于棱镜的SPR系统分析单个细胞外囊泡并确定分子谱以表征从不同细胞分泌的外泌体。
Exosome analysis is a promising tool for clinical and biological research applications. However, detection and biomarker quantification of exosomes is technically challenging because they are small and highly heterogeneous. Here, we report an optical approach for imaging exosomes and quantifying their protein markers without labels using plasmonic scattering microscopy (PSM). PSM can provide improved spatial resolution and distortion-free image compared to conventional surface plasmon resonance (SPR) microscopy, with the signal-to-noise ratio similar to objective coupled surface plasmon resonance (SPR) microscopy, and millimeter-scale field of view as a prism-coupled SPR system, thus allowing exosome size distribution analysis with high throughput. In addition, PSM retains the high specificity and surface sensitivity of the SPR sensors and thus allows selection of exosomes from extracellular vesicles with antibody-modified sensor surfaces and in situ analyzing binding kinetics between antibody and the surface protein biomarkers on the captured exosomes. Finally, the PSM can be easily constructed on a popular prism-coupled SPR system with commercially available components. Thus, it may provide an economical and powerful tool for clinical exosome analysis and exploration of fundamental issues such as exosome biomarker binding properties. The scattering imaging scheme permits the conventional prism-based SPR system to analyse single extracellular vesicles and determine the molecular profiles to characterize the exosomes secreted from different cells.
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