Institute for Personalized Medicine, School of Biomedical Engineering, Shanghai Jiao Tong University, 200030 Shanghai, China.
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, 210093 Nanjing, China.
Proc Natl Acad Sci U S A. 2018 Oct 9;115(41):10275-10280. doi: 10.1073/pnas.1804548115. Epub 2018 Sep 24.
Exosomes play an important role in numerous cellular processes. Fundamental study and practical use of exosomes are significantly constrained by the lack of analytical tools capable of physical and biochemical characterization. In this paper, we present an optical approach capable of imaging single exosomes in a label-free manner, using interferometric plasmonic microscopy. We demonstrate monitoring of the real-time adsorption of exosomes onto a chemically modified Au surface, calculating the image intensity, and determining the size distribution. The sizing capability enables us to quantitatively measure the membrane fusion activity between exosomes and liposomes. We also report the recording of the dynamic interaction between exosomes and antibodies at the single-exosome level, and the tracking of hit-stay-run behavior of exosomes on an antibody-coated surface. We anticipate that the proposed method will contribute to clinical exosome analysis and to the exploration of fundamental issues such as the exosome-antibody binding kinetics.
外泌体在许多细胞过程中发挥着重要作用。由于缺乏能够对其进行物理和生化特性分析的分析工具,对外泌体的基础研究和实际应用受到了显著限制。在本文中,我们提出了一种光学方法,该方法使用干涉等离子体显微镜能够以非标记的方式对单个外泌体进行成像。我们展示了对 exosomes 实时吸附到化学修饰的 Au 表面的监测,计算了图像强度,并确定了尺寸分布。该尺寸测量能力使我们能够定量测量 exosomes 和 liposomes 之间的膜融合活性。我们还报告了在单个外泌体水平上记录 exosomes 和抗体之间的动态相互作用,以及在外泌体在抗体包被表面上的 hit-stay-run 行为的跟踪。我们预计,所提出的方法将有助于临床外泌体分析,并有助于探索外泌体-抗体结合动力学等基本问题。