20.500.12556/RUL-135645 Perspectives of microscopy methods for morphology characterisation of extracellular vesicles from human biofluids 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. extracellular vesicles human biofluids nanotechnology atomic force microscopy electron microscopy morphology true false true Angleški jezik Ni določen Članek v reviji 2022-03-23 12:17:26 2022-03-23 12:17:27 2022-09-14 04:14:26 0000-00-00 00:00:00 2021 0 0 29 str. iss. 6, art. 603 Vol. 9 2021 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 54 2227-9059 10.3390/biomedicines9060603 64802819 RAZ_Malenica_Mladenika_2021.pdf RAZ_Malenica_Mladenika_2021.pdf 1 A78E92DB0C9FE3F110E122233D0EE6AA c3ef85da4365e9a0c6c27a5f94c50790f5cc951cf8f9b3c1587667edb48d1380 09409e34-aa9b-11ec-8aca-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=154852 https://www.mdpi.com/2227-9059/9/6/603 0 e2ceb11f-aa9a-11ec-8aca-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=154851 Medicinska fakulteta 0 0 0