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Imaging of Extracellular Vesicles by Atomic Force Microscopy
Published on: September 11, 2019
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AFM-Based High-Throughput Nanomechanical Screening of Single Extracellular Vesicles
Andrea Ridolfi1,2,3, Marco Brucale1,2, Costanza Montis1,3
1Consorzio Interuniversitario per lo Sviluppo dei Sistemi a Grande Interfase, Via della Lastruccia 3, 50019 Firenze, Italy.
Analytical Chemistry
|July 8, 2020
Summary
This study introduces a rapid atomic force microscopy method to analyze extracellular vesicle (EV) nanomechanics and morphology. The technique allows for high-throughput characterization, distinguishing EVs from contaminants and identifying subtypes based on mechanical properties.
Area of Science:
- Biophysics
- Nanotechnology
- Cell Biology
Background:
- Extracellular vesicles (EVs) possess mechanical properties crucial for their biological functions, including cellular interactions and mechanosensing.
- Traditional methods like force spectroscopy (FS) for measuring individual EV nanomechanics are time-consuming and require specialized expertise.
- A need exists for high-throughput, accessible methods to characterize EV mechanical properties for research and diagnostics.
Purpose of the Study:
- To develop a simplified, high-throughput atomic force microscopy (AFM) procedure for simultaneous nanomechanical and morphological analysis of extracellular vesicles (EVs).
- To enable rapid discrimination of EV subpopulations, contaminants, and subtypes based on mechanical characteristics.
- To validate the method's accuracy and applicability across diverse EV sources.
Main Methods:
- A novel, simplified AFM-based experimental procedure for analyzing hundreds of individual EVs within an hour.
- Utilized basic AFM equipment and freely available software for data acquisition and analysis.
- Simultaneous measurement of nanomechanical properties (stiffness) and morphology of EVs from cell culture, milk, and parasite excretions.
Main Results:
- The method provides a 'nanomechanical snapshot' for discriminating vesicular from non-vesicular objects and EV subtypes.
- Accurate size and stiffness distributions were obtained for EVs from various sources, consistent with traditional FS techniques.
- Successfully detected contamination in EV samples and confirmed the presence of EVs in challenging biological matrices.
Conclusions:
- The developed high-throughput AFM method offers a rapid, accessible, and quantitative approach for EV nanomechanical and morphological analysis.
- This technique is valuable for assessing EV sample purity, identifying subpopulations, and supporting EV isolation protocol development.
- Consistent EV stiffness measurements suggest a potential intrinsic functional role for their mechanical characteristics.
