Molecular rheometry: direct determination of viscosity in Lo and Ld lipid phases via fluorescence lifetime imaging

Yilei Wu, Martin Stefl, Agnieszka Olzynska, Martin Hof, Gokhan Yahioglu, Philip Yip, Duncan R. Casey, Oscar Ces, Jana Humpolickova, Marina K. Kuimova

Research output: Contribution to journalArticle

70 Citations (Scopus)

Abstract

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.
LanguageEnglish
Pages14986-14993
Number of pages8
JournalPhysical Chemistry Chemical Physics
Volume15
Issue number36
DOIs
Publication statusPublished - 9 Jun 2013

Fingerprint

lipids
Fluorescence
Viscosity
viscosity
membranes
Lipids
Imaging techniques
life (durability)
fluorescence
Rotors
Liquids
Spectroscopy
Membrane Lipids
liquids
Membranes
rotors
Cholesterol
cholesterol
Unilamellar Liposomes
Sphingomyelins

Keywords

  • lipid membranes
  • spectroscopy
  • flim
  • phosphatidylcholine

Cite this

Wu, Yilei ; Stefl, Martin ; Olzynska, Agnieszka ; Hof, Martin ; Yahioglu, Gokhan ; Yip, Philip ; Casey, Duncan R. ; Ces, Oscar ; Humpolickova, Jana ; Kuimova, Marina K. / Molecular rheometry : direct determination of viscosity in Lo and Ld lipid phases via fluorescence lifetime imaging. In: Physical Chemistry Chemical Physics. 2013 ; Vol. 15, No. 36. pp. 14986-14993.
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Wu, Y, Stefl, M, Olzynska, A, Hof, M, Yahioglu, G, Yip, P, Casey, DR, Ces, O, Humpolickova, J & Kuimova, MK 2013, 'Molecular rheometry: direct determination of viscosity in Lo and Ld lipid phases via fluorescence lifetime imaging' Physical Chemistry Chemical Physics, vol. 15, no. 36, pp. 14986-14993. https://doi.org/10.1039/c3cp51953h

Molecular rheometry : direct determination of viscosity in Lo and Ld lipid phases via fluorescence lifetime imaging. / Wu, Yilei; Stefl, Martin; Olzynska, Agnieszka; Hof, Martin; Yahioglu, Gokhan ; Yip, Philip; Casey, Duncan R.; Ces, Oscar; Humpolickova, Jana; Kuimova, Marina K.

In: Physical Chemistry Chemical Physics, Vol. 15, No. 36, 09.06.2013, p. 14986-14993.

Research output: Contribution to journalArticle

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AU - Wu, Yilei

AU - Stefl, Martin

AU - Olzynska, Agnieszka

AU - Hof, Martin

AU - Yahioglu, Gokhan

AU - Yip, Philip

AU - Casey, Duncan R.

AU - Ces, Oscar

AU - Humpolickova, Jana

AU - Kuimova, Marina K.

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