An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Valentina Bollati
Università degli Studi di Milano
Corresponding Author
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Extracellular vesicles (EVs) play a key role in many physiological and pathological processes [1]. EVs are a heterogeneous group of membrane-confined particles including endosome-derived exosomes and plasma membrane-originated microvesicles. The expanding field of extracellular vesicle research needs reproducible and accurate methods to characterize EVs [2].
EV profiling can be challenging due to the small size and heterogeneity. This protocol aims to provide a method to isolate EVs and facilitate high-precision particle quantitation by Nanoparticle Tracking Analysis (NTA)[3, 4]. NTA is commonly used to determine EV concentration and diameter [5, 6].
The protocol here described refers to the isolation of EVs from blood-plasma samples by using ultracentrifugation and then quantification and sizing of EVs with NTA by NanoSight NS300 system (Malvern Panalytical Ltd., Malvern, UK) provided with a syringe pump module enabling analysis in constant flow for improved sample statistics.
Keywords
Extracellular Vesicles (EVs), Plasma, Ultracentrifuge, Nanoparticle Tracking Analysis (NTA).
Figure 1
Figure 2
This is a list of supplementary files associated with this preprint. Click to download.
- PlasmaEVsNanosight.avi
Plasma EVs video from Nanosight recording.
Loading...
Version History
CURRENT STATUS: Posted
Version 1
Posted 02 Aug, 2021
No community comments so far
Metrics
Comments: 0
HTML Views: ...
An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Valentina Bollati
Università degli Studi di Milano
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 02 Aug, 2021
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Extracellular vesicles (EVs) play a key role in many physiological and pathological processes [1]. EVs are a heterogeneous group of membrane-confined particles including endosome-derived exosomes and plasma membrane-originated microvesicles. The expanding field of extracellular vesicle research needs reproducible and accurate methods to characterize EVs [2].
EV profiling can be challenging due to the small size and heterogeneity. This protocol aims to provide a method to isolate EVs and facilitate high-precision particle quantitation by Nanoparticle Tracking Analysis (NTA)[3, 4]. NTA is commonly used to determine EV concentration and diameter [5, 6].
The protocol here described refers to the isolation of EVs from blood-plasma samples by using ultracentrifugation and then quantification and sizing of EVs with NTA by NanoSight NS300 system (Malvern Panalytical Ltd., Malvern, UK) provided with a syringe pump module enabling analysis in constant flow for improved sample statistics.
Figure 1
Figure 2
This is a list of supplementary files associated with this preprint. Click to download.
- PlasmaEVsNanosight.avi
Plasma EVs video from Nanosight recording.
Loading...