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Molecular rheometry: direct determination of viscosity in Lo and Ld lipid phases via fluorescence lifetime imaging
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SYSNO ASEP 0394499 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Molecular rheometry: direct determination of viscosity in Lo and Ld lipid phases via fluorescence lifetime imaging Author(s) Wu, Y. (GB)
Štefl, Martin (UFCH-W)
Olžyńska, Agnieszka (UFCH-W) RID
Hof, Martin (UFCH-W) RID, ORCID
Yahioglu, G. (GB)
Yip, P. (GB)
Casey, D. R. (GB)
Ces, O. (GB)
Humpolíčková, Jana (UFCH-W) RID
Kuimova, M. K. (GB)Source Title Physical Chemistry Chemical Physics. - : Royal Society of Chemistry - ISSN 1463-9076
Roč. 15, č. 36 (2013), s. 14986-14993Number of pages 8 s. Language eng - English Country GB - United Kingdom Keywords CORRELATION SPECTROSCOPY ; MODEL MEMBRANES ; LIVE CELLS Subject RIV CF - Physical ; Theoretical Chemistry R&D Projects GBP208/12/G016 GA ČR - Czech Science Foundation (CSF) LH13259 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Institutional support UFCH-W - RVO:61388955 UT WOS 000323520600019 EID SCOPUS 84882963902 DOI 10.1039/c3cp51953h Annotation Understanding of cellular regulatory pathways that involve lipid membranes requires the detailed knowledge of their physical state and structure. However, mapping the viscosity and diffusion in the membranes of complex composition is currently a non-trivial technical challenge. We report fluorescence lifetime spectroscopy and imaging (FLIM) of a meso-substituted BODIPY molecular rotor localised in the leaflet of model membranes of various lipid compositions. We prepare large and giant unilamellar vesicles (LUVs and GUVs) containing phosphatidylcholine (PC) lipids and demonstrate that recording the fluorescence lifetime of the rotor allows us to directly detect the viscosity of the membrane leaflet and to monitor the influence of cholesterol on membrane viscosity in binary and ternary lipid mixtures. In phase-separated 1,2-dioleoyl-sn-glycero-3-phosphocholine-cholesterol–sphingomyelin GUVs we visualise individual liquid ordered (Lo) and liquid disordered (Ld) domains using FLIM and assign specific microscopic viscosities to each domain. Our study showcases the power of FLIM with molecular rotors to image microviscosity of heterogeneous microenvironments in complex biological systems, including membrane-localised lipid rafts. Workplace J. Heyrovsky Institute of Physical Chemistry Contact Michaela Knapová, michaela.knapova@jh-inst.cas.cz, Tel.: 266 053 196 Year of Publishing 2014
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