20.500.12556/RUL-158633 Amphiphilic coumarin-based probes for live-cell STED nanoscopy of plasma membrane Plasma membranes are vital biological structures, serving as protective barriers and participating in various cellular processes. In the field of super-resolution optical microscopy, stimulated emission depletion (STED) nanoscopy has emerged as a powerful method for investigating plasma membrane-related phenomena. However, many applications of STED microscopy are critically restricted by the limited availability of suitable fluorescent probes. This paper reports on the development of two amphiphilic membrane probes, SHE-2H and SHE-2N, specially designed for STED nanoscopy. SHE-2N, in particular, demonstrates quick and stable plasma membrane labelling with negligible intracellular redistribution. Both probes exhibit outstanding photostability and resolution improvement in STED nanoscopy, and are also suited for two-photon excitation microscopy. Furthermore, microscopy experiments and cytotoxicity tests revealed no noticeable cytotoxicity of probe SHE-2N at concentration used for fluorescence imaging. Spectral analysis and fluorescence lifetime measurements conducted on probe SHE-2N using giant unilamellar vesicles, revealed that emission spectra and fluorescence lifetimes exhibited minimal sensitivity to lipid composition variations. These novel probes significantly augment the arsenal of tools available for high-resolution plasma membrane research, enabling a more profound exploration of cellular processes and dynamics. membrane probe fluorescence STED two-photon excitation photostability fizika membrane fluorescenca true false true Angleški jezik Slovenski jezik Članek v reviji 2024-06-18 09:52:01 2024-06-18 09:52:05 2024-07-02 03:35:27 0000-00-00 00:00:00 2024 0 0 10 str. art. 107554 Vol. 150 2024 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 539 0045-2068 10.1016/j.bioorg.2024.107554 198949635 1-s2.0-S0045206824004590-main.pdf 1-s2.0-S0045206824004590-main.pdf 1 230BCC00375357421157120203012EBF fc469b0001e43a072f5506515ce1b4b1e9d9f52866813066c2fabe80fa6acd53 e25223ec-2d47-11ef-88cb-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=186325 https://www.sciencedirect.com/science/article/pii/S0045206824004590 0 bd61b787-2d47-11ef-88cb-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=186324 Fakulteta za farmacijo 0 0 0