Amphiphilic coumarin-based probes for live-cell STED nanoscopy of plasma membrane

Kokot, Hana (author), Kokot, Boštjan (author), Pišlar, Anja (author), Esih, Hana (author), Gabrič, Alen (author), Urbančič, Dunja (author), El, Rojbin (author), Urbančič, Iztok (author), Pajk, Stane (author)

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.

• Language: English
• Keywords: membrane probe, fluorescence, STED, two-photon excitation, photostability
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: FFA - Faculty of Pharmacy
• Publication status: Published
• Publication version: Version of Record
• Year: 2024
• Number of pages: 10 str.
• Numbering: Vol. 150, art. 107554
• PID: 20.500.12556/RUL-158633
• UDC: 539
• ISSN on article: 0045-2068
• DOI: 10.1016/j.bioorg.2024.107554
• COBISS.SI-ID: 198949635
• Publication date in RUL: 18.06.2024

Record is a part of a journal

• Title: Bioorganic chemistry
• Shortened title: Bioorg. chem.
• Publisher: Elsevier
• ISSN: 0045-2068
• COBISS.SI-ID: 25099008

Licences

• License: CC BY-NC 4.0, Creative Commons Attribution-NonCommercial 4.0 International
• Link: http://creativecommons.org/licenses/by-nc/4.0/
• Description: A creative commons license that bans commercial use, but the users don’t have to license their derivative works on the same terms.

Secondary language

• Language: Slovenian
• Keywords: fizika, membrane, fluorescenca