AFM-Based High-Throughput Nanomechanical Screening of Single Extracellular Vesicles

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AFM-Based High-Throughput Nanomechanical Screening of Single Extracellular Vesicles. / Ridolfi, Andrea; Brucale, Marco; Montis, Costanza; Caselli, Lucrezia; Paolini, Lucia; Borup, Anne; Boysen, Anders T; Loria, Francesca; van Herwijnen, Martijn J C; Kleinjan, Marije; Nejsum, Peter; Zarovni, Natasa; Wauben, Marca H M; Berti, Debora; Bergese, Paolo; Valle, Francesco.

In: Analytical Chemistry, Vol. 92, No. 15, 08.2020, p. 10274-10282.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Harvard

Ridolfi, A, Brucale, M, Montis, C, Caselli, L, Paolini, L, Borup, A, Boysen, AT, Loria, F, van Herwijnen, MJC, Kleinjan, M, Nejsum, P, Zarovni, N, Wauben, MHM, Berti, D, Bergese, P & Valle, F 2020, 'AFM-Based High-Throughput Nanomechanical Screening of Single Extracellular Vesicles', Analytical Chemistry, vol. 92, no. 15, pp. 10274-10282. https://doi.org/10.1021/acs.analchem.9b05716

APA

Ridolfi, A., Brucale, M., Montis, C., Caselli, L., Paolini, L., Borup, A., Boysen, A. T., Loria, F., van Herwijnen, M. J. C., Kleinjan, M., Nejsum, P., Zarovni, N., Wauben, M. H. M., Berti, D., Bergese, P., & Valle, F. (2020). AFM-Based High-Throughput Nanomechanical Screening of Single Extracellular Vesicles. Analytical Chemistry, 92(15), 10274-10282. https://doi.org/10.1021/acs.analchem.9b05716

CBE

Ridolfi A, Brucale M, Montis C, Caselli L, Paolini L, Borup A, Boysen AT, Loria F, van Herwijnen MJC, Kleinjan M, Nejsum P, Zarovni N, Wauben MHM, Berti D, Bergese P, Valle F. 2020. AFM-Based High-Throughput Nanomechanical Screening of Single Extracellular Vesicles. Analytical Chemistry. 92(15):10274-10282. https://doi.org/10.1021/acs.analchem.9b05716

MLA

Vancouver

Ridolfi A, Brucale M, Montis C, Caselli L, Paolini L, Borup A et al. AFM-Based High-Throughput Nanomechanical Screening of Single Extracellular Vesicles. Analytical Chemistry. 2020 Aug;92(15):10274-10282. https://doi.org/10.1021/acs.analchem.9b05716

Author

Ridolfi, Andrea ; Brucale, Marco ; Montis, Costanza ; Caselli, Lucrezia ; Paolini, Lucia ; Borup, Anne ; Boysen, Anders T ; Loria, Francesca ; van Herwijnen, Martijn J C ; Kleinjan, Marije ; Nejsum, Peter ; Zarovni, Natasa ; Wauben, Marca H M ; Berti, Debora ; Bergese, Paolo ; Valle, Francesco. / AFM-Based High-Throughput Nanomechanical Screening of Single Extracellular Vesicles. In: Analytical Chemistry. 2020 ; Vol. 92, No. 15. pp. 10274-10282.

Bibtex

@article{bd6e0b13294845758363dc6b5bca5385,
title = "AFM-Based High-Throughput Nanomechanical Screening of Single Extracellular Vesicles",
abstract = "The mechanical properties of extracellular vesicles (EVs) are known to influence their biological function, in terms of, e.g., cellular adhesion, endo/exocytosis, cellular uptake, and mechanosensing. EVs have a characteristic nanomechanical response which can be probed via force spectroscopy (FS) and exploited to single them out from nonvesicular contaminants or to discriminate between subtypes. However, measuring the nanomechanical characteristics of individual EVs via FS is a labor-intensive and time-consuming task, usually limiting this approach to specialists. Herein, we describe a simple atomic force microscopy based experimental procedure for the simultaneous nanomechanical and morphological analysis of several hundred individual nanosized EVs within the hour time scale, using basic AFM equipment and skills and only needing freely available software for data analysis. This procedure yields a {"}nanomechanical snapshot{"} of an EV sample which can be used to discriminate between subpopulations of vesicular and nonvesicular objects in the same sample and between populations of vesicles with similar sizes but different mechanical characteristics. We demonstrate the applicability of the proposed approach to EVs obtained from three very different sources (human colorectal carcinoma cell culture, raw bovine milk, and Ascaris suum nematode excretions), recovering size and stiffness distributions of individual vesicles in a sample. EV stiffness values measured with our high-throughput method are in very good quantitative accord with values obtained by FS techniques which measure EVs one at a time. We show how our procedure can detect EV samples contamination by nonvesicular aggregates and how it can quickly attest the presence of EVs even in samples for which no established assays and/or commercial kits are available (e.g., Ascaris EVs), thus making it a valuable tool for the rapid assessment of EV samples during the development of isolation/enrichment protocols by EV researchers. As a side observation, we show that all measured EVs have a strikingly similar stiffness, further reinforcing the hypothesis that their mechanical characteristics could have a functional role.",
author = "Andrea Ridolfi and Marco Brucale and Costanza Montis and Lucrezia Caselli and Lucia Paolini and Anne Borup and Boysen, {Anders T} and Francesca Loria and {van Herwijnen}, {Martijn J C} and Marije Kleinjan and Peter Nejsum and Natasa Zarovni and Wauben, {Marca H M} and Debora Berti and Paolo Bergese and Francesco Valle",
year = "2020",
month = aug,
doi = "10.1021/acs.analchem.9b05716",
language = "English",
volume = "92",
pages = "10274--10282",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "AMER CHEMICAL SOC",
number = "15",

}

RIS

TY - JOUR

T1 - AFM-Based High-Throughput Nanomechanical Screening of Single Extracellular Vesicles

AU - Ridolfi, Andrea

AU - Brucale, Marco

AU - Montis, Costanza

AU - Caselli, Lucrezia

AU - Paolini, Lucia

AU - Borup, Anne

AU - Boysen, Anders T

AU - Loria, Francesca

AU - van Herwijnen, Martijn J C

AU - Kleinjan, Marije

AU - Nejsum, Peter

AU - Zarovni, Natasa

AU - Wauben, Marca H M

AU - Berti, Debora

AU - Bergese, Paolo

AU - Valle, Francesco

PY - 2020/8

Y1 - 2020/8

N2 - The mechanical properties of extracellular vesicles (EVs) are known to influence their biological function, in terms of, e.g., cellular adhesion, endo/exocytosis, cellular uptake, and mechanosensing. EVs have a characteristic nanomechanical response which can be probed via force spectroscopy (FS) and exploited to single them out from nonvesicular contaminants or to discriminate between subtypes. However, measuring the nanomechanical characteristics of individual EVs via FS is a labor-intensive and time-consuming task, usually limiting this approach to specialists. Herein, we describe a simple atomic force microscopy based experimental procedure for the simultaneous nanomechanical and morphological analysis of several hundred individual nanosized EVs within the hour time scale, using basic AFM equipment and skills and only needing freely available software for data analysis. This procedure yields a "nanomechanical snapshot" of an EV sample which can be used to discriminate between subpopulations of vesicular and nonvesicular objects in the same sample and between populations of vesicles with similar sizes but different mechanical characteristics. We demonstrate the applicability of the proposed approach to EVs obtained from three very different sources (human colorectal carcinoma cell culture, raw bovine milk, and Ascaris suum nematode excretions), recovering size and stiffness distributions of individual vesicles in a sample. EV stiffness values measured with our high-throughput method are in very good quantitative accord with values obtained by FS techniques which measure EVs one at a time. We show how our procedure can detect EV samples contamination by nonvesicular aggregates and how it can quickly attest the presence of EVs even in samples for which no established assays and/or commercial kits are available (e.g., Ascaris EVs), thus making it a valuable tool for the rapid assessment of EV samples during the development of isolation/enrichment protocols by EV researchers. As a side observation, we show that all measured EVs have a strikingly similar stiffness, further reinforcing the hypothesis that their mechanical characteristics could have a functional role.

AB - The mechanical properties of extracellular vesicles (EVs) are known to influence their biological function, in terms of, e.g., cellular adhesion, endo/exocytosis, cellular uptake, and mechanosensing. EVs have a characteristic nanomechanical response which can be probed via force spectroscopy (FS) and exploited to single them out from nonvesicular contaminants or to discriminate between subtypes. However, measuring the nanomechanical characteristics of individual EVs via FS is a labor-intensive and time-consuming task, usually limiting this approach to specialists. Herein, we describe a simple atomic force microscopy based experimental procedure for the simultaneous nanomechanical and morphological analysis of several hundred individual nanosized EVs within the hour time scale, using basic AFM equipment and skills and only needing freely available software for data analysis. This procedure yields a "nanomechanical snapshot" of an EV sample which can be used to discriminate between subpopulations of vesicular and nonvesicular objects in the same sample and between populations of vesicles with similar sizes but different mechanical characteristics. We demonstrate the applicability of the proposed approach to EVs obtained from three very different sources (human colorectal carcinoma cell culture, raw bovine milk, and Ascaris suum nematode excretions), recovering size and stiffness distributions of individual vesicles in a sample. EV stiffness values measured with our high-throughput method are in very good quantitative accord with values obtained by FS techniques which measure EVs one at a time. We show how our procedure can detect EV samples contamination by nonvesicular aggregates and how it can quickly attest the presence of EVs even in samples for which no established assays and/or commercial kits are available (e.g., Ascaris EVs), thus making it a valuable tool for the rapid assessment of EV samples during the development of isolation/enrichment protocols by EV researchers. As a side observation, we show that all measured EVs have a strikingly similar stiffness, further reinforcing the hypothesis that their mechanical characteristics could have a functional role.

U2 - 10.1021/acs.analchem.9b05716

DO - 10.1021/acs.analchem.9b05716

M3 - Journal article

C2 - 32631050

VL - 92

SP - 10274

EP - 10282

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 15

ER -