Year: 2025
Type: article
Abstract: The "stickiness" of extracellular vesicles (EVs) can pose challenges for EV processing and storage, but adhesive properties may also be exploited to immobilize EVs directly on surfaces for various measurement techniques, including super-resolution microscopy. Direct adhesion to surfaces may allow examination of broader populations of EVs than molecular affinity approaches, which can also involve specialized, expensive affinity reagents. Here, we report on the interaction of EVs with borosilicate glass and quartz coverslips and on the effects of pre-coating coverslips with poly-L-lysine (PLL), a reagent commonly used to facilitate interactions between negatively charged surfaces of cells and amorphous surfaces. Additionally, we compared two mounting media conditions for super-resolution microscopy (SRM) imaging and used immobilized EVs for a B-cell interaction test. Our findings suggest that borosilicate glass coverslips immobilize EVs better than quartz glass coverslips. We also found that PLL is not strictly required for EV retention but contributes to the uniform distribution of EVs on borosilicate glass coverslips. Overall, these findings suggest that standard lab materials like borosilicate glass coverslips, with or without PLL, can be effectively used for the immobilization of EVs in specific imaging techniques.
Source: Journal of Extracellular Biology
DOI: 10.1002/jex2.70042
Institutions: Johns Hopkins University, Universidade Federal de São Carlos
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $3,260
Awards: G1936050780, G2305452004, G295657101, G3466191908, G3719139961, G4204137714, Novel Approaches to Capture, Sorting, and Characterization of CNS-Origin Extracellular Vesicles, G5586281196, G6438247056, Novel Separation Methods for exRNA Carriers: Extracellular Vesicles, Lipoprotein Particles, and Protein Aggregates, Novel Separation Methods for exRNA Carriers: Extracellular Vesicles, Lipoprotein Particles, and Protein Aggregates, Next-generation extracellular vesicle biologics to target central nervous system and peripheral reservoirs of HIV, G9000156707, G9000203527, Continued support for an SPF Macaca nemestrina breeding colony to provide animals to NIH funded HIV research projects, G905798045, Novel Approaches to Capture, Sorting, and Characterization of CNS-Origin Extracellular Vesicles
OpenAlex ID: W4409795429