Exploring the Adhesion Properties of Extracellular Vesicles for Functional Assays

ABSTRACT The ‛stickiness’ of extracellular vesicles (EVs) can pose challenges for EV processing and storage, but adhesive properties may also be exploited to immobilise EVs directly on surfaces for various measurement techniques, including super‐resolution microscopy (SRM). Direct adhesion to surfac...

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Main Authors: Bianca C. Pachane, Bess Carlson, Suzanne E. Queen, Heloisa S. Selistre‐de‐Araujo, Kenneth W. Witwer
Format: Article
Language:English
Published: Wiley 2025-04-01
Series:Journal of Extracellular Biology
Subjects:
Online Access:https://doi.org/10.1002/jex2.70042
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author Bianca C. Pachane
Bess Carlson
Suzanne E. Queen
Heloisa S. Selistre‐de‐Araujo
Kenneth W. Witwer
author_facet Bianca C. Pachane
Bess Carlson
Suzanne E. Queen
Heloisa S. Selistre‐de‐Araujo
Kenneth W. Witwer
author_sort Bianca C. Pachane
collection DOAJ
description ABSTRACT The ‛stickiness’ of extracellular vesicles (EVs) can pose challenges for EV processing and storage, but adhesive properties may also be exploited to immobilise EVs directly on surfaces for various measurement techniques, including super‐resolution microscopy (SRM). Direct adhesion to surfaces may allow the examination of broader populations of EVs than molecular affinity approaches, which can also involve specialised, 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 SRM imaging and used immobilised EVs for a B‐cell interaction test. Our findings suggest that borosilicate glass coverslips immobilise 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 immobilisation of EVs in specific imaging techniques.
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publishDate 2025-04-01
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spelling doaj-art-653299a75bfa44aea5c49f519f41e3e12025-08-20T02:29:46ZengWileyJournal of Extracellular Biology2768-28112025-04-0144n/an/a10.1002/jex2.70042Exploring the Adhesion Properties of Extracellular Vesicles for Functional AssaysBianca C. Pachane0Bess Carlson1Suzanne E. Queen2Heloisa S. Selistre‐de‐Araujo3Kenneth W. Witwer4Department of Molecular and Comparative Pathobiology Johns Hopkins University School of Medicine Baltimore Maryland USADepartment of Molecular and Comparative Pathobiology Johns Hopkins University School of Medicine Baltimore Maryland USADepartment of Molecular and Comparative Pathobiology Johns Hopkins University School of Medicine Baltimore Maryland USABiochemistry and Molecular Biology Laboratory, Department of Physiological Sciences Universidade Federal de São Carlos – UFSCar São Carlos São Paulo BrazilDepartment of Molecular and Comparative Pathobiology Johns Hopkins University School of Medicine Baltimore Maryland USAABSTRACT The ‛stickiness’ of extracellular vesicles (EVs) can pose challenges for EV processing and storage, but adhesive properties may also be exploited to immobilise EVs directly on surfaces for various measurement techniques, including super‐resolution microscopy (SRM). Direct adhesion to surfaces may allow the examination of broader populations of EVs than molecular affinity approaches, which can also involve specialised, 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 SRM imaging and used immobilised EVs for a B‐cell interaction test. Our findings suggest that borosilicate glass coverslips immobilise 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 immobilisation of EVs in specific imaging techniques.https://doi.org/10.1002/jex2.70042adhesionextracellular vesiclessuper‐resolution microscopy
spellingShingle Bianca C. Pachane
Bess Carlson
Suzanne E. Queen
Heloisa S. Selistre‐de‐Araujo
Kenneth W. Witwer
Exploring the Adhesion Properties of Extracellular Vesicles for Functional Assays
Journal of Extracellular Biology
adhesion
extracellular vesicles
super‐resolution microscopy
title Exploring the Adhesion Properties of Extracellular Vesicles for Functional Assays
title_full Exploring the Adhesion Properties of Extracellular Vesicles for Functional Assays
title_fullStr Exploring the Adhesion Properties of Extracellular Vesicles for Functional Assays
title_full_unstemmed Exploring the Adhesion Properties of Extracellular Vesicles for Functional Assays
title_short Exploring the Adhesion Properties of Extracellular Vesicles for Functional Assays
title_sort exploring the adhesion properties of extracellular vesicles for functional assays
topic adhesion
extracellular vesicles
super‐resolution microscopy
url https://doi.org/10.1002/jex2.70042
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AT suzanneequeen exploringtheadhesionpropertiesofextracellularvesiclesforfunctionalassays
AT heloisasselistredearaujo exploringtheadhesionpropertiesofextracellularvesiclesforfunctionalassays
AT kennethwwitwer exploringtheadhesionpropertiesofextracellularvesiclesforfunctionalassays