Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 506954
Title Single Molecule 3D Orientation in Time and Space : A 6D Dynamic Study on Fluorescently Labeled Lipid Membranes
Author(s) Börner, Richard; Ehrlich, Nicky; Hohlbein, Johannes; Hübner, Christian G.
Source Journal of Fluorescence 26 (2016)3. - ISSN 1053-0509 - p. 963 - 975.
DOI https://doi.org/10.1007/s10895-016-1784-5
Department(s) Biophysics
EPS
Publication type Refereed Article in a scientific journal
Publication year 2016
Keyword(s) 3D dipole orientation - Anisotropy - Confocal microscopy - FCS - Fluorescence microscopy - GUV - Single molecule
Abstract

Interactions between single molecules profoundly depend on their mutual three-dimensional orientation. Recently, we demonstrated a technique that allows for orientation determination of single dipole emitters using a polarization-resolved distribution of fluorescence into several detection channels. As the method is based on the detection of single photons, it additionally allows for performing fluorescence correlation spectroscopy (FCS) as well as dynamical anisotropy measurements thereby providing access to fast orientational dynamics down to the nanosecond time scale. The 3D orientation is particularly interesting in non-isotropic environments such as lipid membranes, which are of great importance in biology. We used giant unilamellar vesicles (GUVs) labeled with fluorescent dyes down to a single molecule concentration as a model system for both, assessing the robustness of the orientation determination at different timescales and quantifying the associated errors. The vesicles provide a well-defined spherical surface, such that the use of fluorescent lipid dyes (DiO) allows to establish a a wide range of dipole orientations experimentally. To complement our experimental data, we performed Monte Carlo simulations of the rotational dynamics of dipoles incorporated into lipid membranes. Our study offers a comprehensive view on the dye orientation behavior in a lipid membrane with high spatiotemporal resolution representing a six-dimensional fluorescence detection approach.

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