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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Official URL: https://www.mdpi.com/2227-9059/9/6/603

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 title	Project leader	Project code	Project type
Identifikacija cirkulirajućih biomarkera neurološkog oporavka u bolesnika s ozljedom mozga-CiMozak / CirBrain	Kristina Grabušić	IP-2019-04-1511	HRZZ
Razvoj istraživačke infrastrukture na kampusu Sveučilišta u Rijeci-RISK	Nevenka Ožanić	RC.2.2.06-0001	EK
Uspostava protokola na Sveučilištu u Rijeci za primjenu nanotehnoloških metoda: mikroskopije atomskih sila i skenirajućeg elektronskog mikroskopa prilikom vizualizacije i karakterizacije egzosoma-	Mladenka Malenica Staver	uniri-biomed-18-279	NadSve
Izvanstanične vezikule kao klinički markeri neuroregeneracije nakon teške ozljede mozga-	Kristina Grabušić	uniri-biomed-18-5	NadSve
Analiza novih kompozitnih materijala i biljnih ekstrakata spektroskopskim tehnikama-	Srećko Valić	uniri-tehnic-18-294	NadSve