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Volume 9
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10.3390/biomedicines9060603
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Perspectives of Microscopy Methods for Morphology Characterisation of Extracellular Vesicles from Human Biofluids

by
Mladenka Malenica
1,*,†,
Marija Vukomanović
2,†,
Mario Kurtjak
2,
Valentina Masciotti
3,
Simone dal Zilio
3,
Silvio Greco
4,
Marco Lazzarino
3,
Vedrana Krušić
1,
Marko Perčić
5,6,
Ivana Jelovica Badovinac
6,7,
Karmen Wechtersbach
8,
Ivona Vidović
9,
Vanja Baričević
9,
Srećko Valić
10,11,
Pero Lučin
1,
Nika Kojc
8 and
Kristina Grabušić
1
1
Department of Physiology and Immunology, Faculty of Medicine, University of Rijeka, HR-51000 Rijeka, Croatia
2
Advanced Materials Department, Jožef Stefan Institute, SI-1000 Ljubljana, Slovenia
3
CNR-IOM Istituto Officina dei Materiali-Consiglio Nazionale delle Ricerche c/Area Scinece Park, Basovizza, I-34149 Trieste, Italy
4
A. P. E. Research srl, I-34151 Trieste, Italy
5
Faculty of Engineering, University of Rijeka, HR-51000 Rijeka, Croatia
6
Centre for Micro- and Nanosciences and Technologies, University of Rijeka, HR-51000 Rijeka, Croatia
7
Department of Physics, University of Rijeka, HR-51000 Rijeka, Croatia
8
Faculty of Medicine, Institute of Pathology, University of Ljubljana, SI-1000 Ljubljana, Slovenia
9
Department of Biotechnology, University of Rijeka, HR-51000 Rijeka, Croatia
10
Department of Medical Chemistry, Biochemistry and Clinical Chemistry, Faculty of Medicine, University of Rijeka, HR-51000 Rijeka, Croatia
11
Division of Physical Chemistry, Ruđer Bošković Institute, HR-10000 Zagreb, Croatia
 
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*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Academic Editor: Stefania Raimondo
Biomedicines 2021, 9(6), 603; https://doi.org/10.3390/biomedicines9060603
Received: 30 April 2021 / Revised: 18 May 2021 / Accepted: 21 May 2021 / Published: 26 May 2021
(This article belongs to the Special Issue Feature Papers in "Biomedical Materials and Nanomedicine")
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Abstract

Extracellular vesicles (EVs) are nanometric membranous structures secreted from almost every cell and present in biofluids. Because EV composition reflects the state of its parental tissue, EVs possess an enormous diagnostic/prognostic potential to reveal pathophysiological conditions. However, a prerequisite for such usage of EVs is their detailed characterisation, including visualisation which is mainly achieved by atomic force microscopy (AFM) and electron microscopy (EM). Here we summarise the EV preparation protocols for AFM and EM bringing out the main challenges in the imaging of EVs, both in their natural environment as biofluid constituents and in a saline solution after EV isolation. In addition, we discuss approaches for EV imaging and identify the potential benefits and disadvantages when different AFM and EM methods are applied, including numerous factors that influence the morphological characterisation, standardisation, or formation of artefacts. We also demonstrate the effects of some of these factors by using cerebrospinal fluid as an example of human biofluid with a simpler composition. Here presented comparison of approaches to EV imaging should help to estimate the current state in morphology research of EVs from human biofluids and to identify the most efficient pathways towards the standardisation of sample preparation and microscopy modes. View Full-Text
Keywords: extracellular vesicles; human biofluids; nanotechnology; atomic force microscopy; electron microscopy; morphology extracellular vesicles; human biofluids; nanotechnology; atomic force microscopy; electron microscopy; morphology
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MDPI and ACS Style

Malenica, M.; Vukomanović, M.; Kurtjak, M.; Masciotti, V.; dal Zilio, S.; Greco, S.; Lazzarino, M.; Krušić, V.; Perčić, M.; Jelovica Badovinac, I.; Wechtersbach, K.; Vidović, I.; Baričević, V.; Valić, S.; Lučin, P.; Kojc, N.; Grabušić, K. Perspectives of Microscopy Methods for Morphology Characterisation of Extracellular Vesicles from Human Biofluids. Biomedicines 2021, 9, 603. https://doi.org/10.3390/biomedicines9060603

AMA Style

Malenica M, Vukomanović M, Kurtjak M, Masciotti V, dal Zilio S, Greco S, Lazzarino M, Krušić V, Perčić M, Jelovica Badovinac I, Wechtersbach K, Vidović I, Baričević V, Valić S, Lučin P, Kojc N, Grabušić K. Perspectives of Microscopy Methods for Morphology Characterisation of Extracellular Vesicles from Human Biofluids. Biomedicines. 2021; 9(6):603. https://doi.org/10.3390/biomedicines9060603

Chicago/Turabian Style

Malenica, Mladenka, Marija Vukomanović, Mario Kurtjak, Valentina Masciotti, Simone dal Zilio, Silvio Greco, Marco Lazzarino, Vedrana Krušić, Marko Perčić, Ivana Jelovica Badovinac, Karmen Wechtersbach, Ivona Vidović, Vanja Baričević, Srećko Valić, Pero Lučin, Nika Kojc, and Kristina Grabušić. 2021. "Perspectives of Microscopy Methods for Morphology Characterisation of Extracellular Vesicles from Human Biofluids" Biomedicines 9, no. 6: 603. https://doi.org/10.3390/biomedicines9060603

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