Decoding how cells communicate by high-speed imaging of molecular couriers
Nano Life Science Institute (WPI-NanoLSI), Kanazawa University, demonstrates a novel approach for nanoscopic profiling of small extracellular vesicles (sEVs) using high-speed atomic force microscopy (HS-AFM) videography. This pioneering method provides an unprecedented level of detail in characterizing sEV subpopulations, offering new insights into their biological roles and potential applications in disease diagnostics.
Extracellular vesicles (EVs), including exosomes and microvesicles, play a crucial role in intercellular communication by transferring biomolecules such as proteins, lipids, and RNA. These tiny vesicles act as molecular couriers, shuttling messages between cells to regulate immune responses, tissue repair, and even disease progression. Despite their significance, the accurate characterization of these nanoscale vesicles remains a major challenge due to their small size and heterogeneity. Existing techniques, such as nanoparticle tracking analysis (NTA) and flow cytometry, lack the resolution to provide precise structural and compositional information at the single-vesicle level.
In this study, led by Keesiang Lim and Richard W. Wong, the team utilized HS-AFM to visualize the nanotopology of sEVs derived from HEK293T cells under physiological conditions. Their findings revealed distinct sEV subpopulations enriched with specific exosome markers, such as CD63 and CD81. Notably, they observed that sEVs smaller than 100 nm exhibited greater membrane rigidity and higher co-localization with exosomal markers compared to larger vesicles, which showed significant height fluctuations.
“Our study represents a major advancement in extracellular vesicle research,” said Wong. “By leveraging HS-AFM videography, we can now directly observe the dynamic interactions of surface markers on individual sEVs, paving the way for the development of high-precision EV-based biomarkers.”
This nanoscopic immunophenotyping approach has the potential to revolutionize early disease detection, particularly in cancer diagnostics, where exosome-based biomarkers are gaining attention. Additionally, the method could contribute to advancements in targeted drug delivery and regenerative medicine by enabling more precise characterization of therapeutic EVs.
Glossary
Molecular Couriers: Small extracellular vesicles (sEVs) that transport biomolecules between cells, facilitating intercellular communication.
High-Speed Atomic Force Microscopy (HS-AFM): A high-resolution imaging technique that enables real-time visualization of nanoscale structures and dynamic biological processes.
Nanotopology: The study of surface structures and nanoscale features of biological entities, such as extracellular vesicles, under physiological conditions.
Immunophenotyping: The process of identifying specific biomolecular markers on cell-derived vesicles using antibodies to differentiate subpopulations.
Article
- Title
- Nanoscopic Profiling of Small Extracellular Vesicles via High-Speed Atomic Force Microscopy (HS-AFM) Videography
- Author
- Muhammad Isman Sandira, Keesiang Lim, Takeshi Yoshida, Elma Sakinatus Sajidah, Shinnosuke Narimatsu, Reon Imakawa, Kota Yoshimura, Goro Nishide, Yujia Qiu, Azuma Taoka, Masaharu Hazawa, Toshio Ando, Rikinari Hanayama, Richard W Wong
- Journal
- Journal of Extracellular Vesicles
- Publication date
- Mar 26, 2025
- DOI
- 10.1002/jev2.70050
- URL
- https://isevjournals.onlinelibrary.wiley.com/doi/full/10.1002/jev2.70050