Nanoscopic Profiling of Small Extracellular Vesicles via High-Speed Atomic Force Microscopy (HS-AFM) Videography

Abstract

100 nm) exhibited greater height fluctuations compared to smaller sEVs (diameter ≤ 100 nm). Next, we found that mouse-origin IgGCD63, and rabbit-origin IgGcontrol and IgGCD81, exhibited the iconic ‘Y’ conformation, and similar structural dynamics properties. Last, exosome marker antibodies predominantly co-localised with sEVd ≤ 100 nm but not with sEVd > 100 nm, demonstrating the CD63-CD81-enriched sEV and CD63-CD81-depleted sEV subpopulations. In summary, we demonstrate that nanoscopic profiling of surface exosome markers on sEVs using HS-AFM is feasible for characterising distinct sEV subpopulations in a heterogeneous sEV mixture.