20.500.12556/DKUM-90244 How to control fluorescent labeling of metal oxide nanoparticles for artefact-free live cell microscopy Nanotechnologies hold great promise for various applications. To predict and guarantee the safety of novel nanomaterials, it is essential to understand their mechanism of action in an organism, causally connecting adverse outcomes with early molecular events. This is best investigated using noninvasive advanced optical methods, such as high-resolution live-cell fluorescence microscopy, which require stable labeling of nanoparticles with fluorescent dyes. However, as shown here, when the labeling is performed inadequately, unbound fluorescent dyes and inadvertently altered chemical and physical properties of the nanoparticles can result in experimental artefacts and erroneous conclusions. To prevent such unintentional errors, we introduce a tested minimal combination of experimental methods to enable artefact-free fluorescent labeling of metal-oxide nanoparticles—the largest subpopulation of nanoparticles by industrial production and applications—and demonstrate its application in the case of TiO2 nanotubes. We (1) characterize potential changes of the nanoparticles’ surface charge and morphology that might occur during labeling by using zeta potential measurements and transmission electron microscopy, respectively, and (2) assess stable binding of the fluorescent dye to the nanoparticles with either fluorescence intensity measurements or fluorescence correlation spectroscopy, which ensures correct nanoparticle localization. Together, these steps warrant the reliability and reproducibility of advanced optical tracking, which is necessary to explore nanomaterials’ mechanism of action and will foster widespread and safe use of new nanomaterials. dye leakage labeling characterization artefacts labeling-induced changes reliable nanomaterial labeling uhajanje barvila karakterizacija označevanja artefakti spremembe zaradi označevanja zanesljivo označevanje nanomaterialov true false true Informa Healthcare Angleški jezik Slovenski jezik Znanstveno delo 2024-08-27 12:13:27 2024-08-27 12:41:58 2024-08-28 03:19:35 0000-00-00 00:00:00 2021 0 0 Str. 1102-1123 Št. 8 Letn. 15 2021 0000-00-00 Zaloznikova Objavljeno NiDoloceno 2021-06-11 2021-08-23 2021-10-06 53 79746819 10.1080/17435390.2021.1973607 1743-5404 Kokot-2021-How_to_control_fluorescent_labeling.pdf Kokot-2021-How_to_control_fluorescent_labeling.pdf 1 2536085C334BEFF263F687ABD02D1570 3f67c9a1880eef1972931eac19fa891e7cc77139d55b8190ed493be14d172d80 ba14d63d-6460-11ef-af0b-00155d000105 https://dk.um.si/Dokument.php?lang=slv&id=183447 https://doi.org/10.1080/17435390.2021.1973607 0 c49f37af-6460-11ef-af0b-00155d000105 https://dk.um.si/Dokument.php?lang=slv&id=183448 Fakulteta za naravoslovje in matematiko 0 0 0