Isogai Tatsuki, Hirosawa Koichiro M, Suzuki Kenichi G N
The United Graduate School of Agricultural Science, Gifu University, Gifu 501-1193, Japan.
Institute for Glyco-Core Research (iGCORE), Gifu University, Gifu 501-1193, Japan.
Molecules. 2024 Dec 10;29(24):5828. doi: 10.3390/molecules29245828.
Extracellular vesicles (EVs), secreted from most cells, are small lipid membranes of vesicles of 30 to 1000 nm in diameter and contain nucleic acids, proteins, and intracellular organelles originating from donor cells. EVs play pivotal roles in intercellular communication, particularly in forming niches for cancer cell metastasis. However, EVs derived from donor cells exhibit significant heterogeneity, complicating the investigation of EV subtypes using ensemble averaging methods. In this context, we highlight recent studies that characterize individual EVs using advanced techniques, including single-fluorescent-particle tracking, single-metal-nanoparticle tracking, single-non-label-particle tracking, super-resolution microscopy, and atomic force microscopy. These techniques have facilitated high-throughput analyses of the properties of individual EV particles such as their sizes, compositions, and physical properties. Finally, we address the challenges that need to be resolved via single-particle (-molecule) imaging and super-resolution microscopy in future research.
细胞外囊泡（EVs）由大多数细胞分泌，是直径为30至1000纳米的小脂质膜囊泡，包含源自供体细胞的核酸、蛋白质和细胞内细胞器。EVs在细胞间通讯中起关键作用，尤其是在为癌细胞转移形成微环境方面。然而，源自供体细胞的EVs表现出显著的异质性，这使得使用总体平均方法研究EV亚型变得复杂。在此背景下，我们重点介绍了最近使用先进技术对单个EVs进行表征的研究，这些技术包括单荧光粒子追踪、单金属纳米粒子追踪、单无标记粒子追踪、超分辨率显微镜和原子力显微镜。这些技术促进了对单个EV粒子特性（如大小、组成和物理性质）的高通量分析。最后，我们阐述了在未来研究中通过单粒子（-分子）成像和超分辨率显微镜需要解决的挑战。
