5 March 2015 Investigating cell membrane structure and dynamics with TCSPC-FLIM
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Abstract
We report the use of Time-Correlated Single Photon Counting (TCSPC) in a polarization-resolved Fluorescence Lifetime Imaging (FLIM) setup for the investigation of cell membrane structural and dynamic properties. This technique allows us to study the orientation and mobility of fluorescent membrane dyes, namely di-4-ANEPPDHQ and DiO, in model bilayers of different lipid compositions. Dipole alignment and extent of rotational motion can be linked to membrane order and fluidity. Comparison of the time-resolved anisotropy decays of the two fluorescent dyes suggests that rotational motion of membrane constituents is restricted in liquid-ordered phases, and appears to be limited to the region of aliphatic tails in liquid-disordered phases. In living cells, understanding the membrane structure provides crucial information on its functional properties, such as exo- and endocytosis, cell mobility and signal transduction.
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Alix Le Marois, Dylan M. Owen, Klaus Suhling, "Investigating cell membrane structure and dynamics with TCSPC-FLIM", Proc. SPIE 9329, Multiphoton Microscopy in the Biomedical Sciences XV, 932938 (5 March 2015); doi: 10.1117/12.2178563; https://doi.org/10.1117/12.2178563
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