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Perspectives of Microscopy Methods for Morphology Characterisation of Extracellular Vesicles from Human Biofluids

Malenica, Mladenka; Vukomanović, Marija; Kurtjak, Mario; Masciotti, Valentina; dal Zilio, Simone; Greco, Silvio; Lazzarino, Marco; Krušić, Vedrana; Perčić, Marko; Jelovica Badovinac, Ivana; Wechtersbach, Karmen; Vidović, Ivona; Baričević, Vanja; Valić, Srećko; Lučin, Pero; Kojc, Nika; Grabušić, Kristina (2021) Perspectives of Microscopy Methods for Morphology Characterisation of Extracellular Vesicles from Human Biofluids. Biomedicines, 9 (6). ISSN 2227-9059

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Abstract

Extracellular vesicles (EVs) are nanometric membranous structures secreted from almostevery 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 visuali-sation 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 challengesin the imaging of EVs, both in their natural environment as biofluid constituents and in a salinesolution after EV isolation. In addition, we discuss approaches for EV imaging and identify thepotential benefits and disadvantages when different AFM and EM methods are applied, includingnumerous factors that influence the morphological characterisation, standardisation, or formationof artefacts. We also demonstrate the effects of some of these factors by using cerebrospinal fluidas an example of human biofluid with a simpler composition. Here presented comparison of ap-proaches to EV imaging should help to estimate the current state in morphology research of EVsfrom human biofluids and to identify the most efficient pathways towards the standardisation ofsample preparation and microscopy modes.

Item Type: Article
Uncontrolled Keywords: extracellular vesicles ; human biofluids ; nanotechnology ; atomic force microscopy ; electron microscopy ; morphology
Subjects: INTERDISCIPLINARY AREAS OF KNOWLEDGE > Biotechnology in Biomedicine (natural science, biomedicine and healthcare, bioethics area
Divisions: Division of Physical Chemistry
Projects:
Project titleProject leaderProject codeProject type
Identifikacija cirkulirajućih biomarkera neurološkog oporavka u bolesnika s ozljedom mozga-CiMozak / CirBrainKristina GrabušićIP-2019-04-1511HRZZ
Razvoj istraživačke infrastrukture na kampusu Sveučilišta u Rijeci-RISKNevenka OžanićRC.2.2.06-0001EK
Uspostava protokola na Sveučilištu u Rijeci za primjenu nanotehnoloških metoda: mikroskopije atomskih sila i skenirajućeg elektronskog mikroskopa prilikom vizualizacije i karakterizacije egzosoma-Mladenka Malenica Staveruniri-biomed-18-279NadSve
Izvanstanične vezikule kao klinički markeri neuroregeneracije nakon teške ozljede mozga-Kristina Grabušićuniri-biomed-18-5NadSve
Analiza novih kompozitnih materijala i biljnih ekstrakata spektroskopskim tehnikama-Srećko Valićuniri-tehnic-18-294NadSve
Depositing User: Srećko Valić
Date Deposited: 28 Jun 2021 12:38
Last Modified: 06 Aug 2021 12:39
URI: http://fulir.irb.hr/id/eprint/6461
DOI: 10.3390/biomedicines9060603

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