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Membranes 2015, 5(4), 702-721; doi:10.3390/membranes5040702

Lipid Diffusion in Supported Lipid Bilayers: A Comparison between Line-Scanning Fluorescence Correlation Spectroscopy and Single-Particle Tracking

1
Department of Physics and Astronomy, McMaster University, Hamilton, ON L8S 4M1, Canada
2
Department of Chemistry and Chemical Biology, McMaster University, Hamilton, ON L8S 4L8, Canada
3
Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON L8N 3Z5, Canada
These authors contributed equally to this work.
*
Authors to whom correspondence should be addressed.
Academic Editor: Maikel Rheinstadter
Received: 17 October 2015 / Accepted: 6 November 2015 / Published: 13 November 2015
(This article belongs to the Special Issue Membrane Structure and Dynamics)
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Abstract

Diffusion in lipid membranes is an essential component of many cellular process and fluorescence a method of choice to study membrane dynamics. The goal of this work was to directly compare two common fluorescence methods, line-scanning fluorescence correlation spectroscopy and single-particle tracking, to observe the diffusion of a fluorescent lipophilic dye, DiD, in a complex five-component mitochondria-like solid-supported lipid bilayer. We measured diffusion coefficients of \(D_{\text{FCS}} \sim\) 3 \(μ\text{m}^2\cdot\text{s}^{-1}\) and \(D_{\text{SPT}} \sim\) 2 \( μ\text{m}^2\cdot\text{s}^{-1}\), respectively. These comparable, yet statistically different values are used to highlight the main message of the paper, namely that the two considered methods give access to distinctly different dynamic ranges: \(D \gtrsim\) 1 \(μ\text{m}^2\cdot\text{s}^{-1}\) for FCS and \(D \lesssim\) 5 \(μ\text{m}^2\cdot\text{s}^{-1}\) for SPT (with standard imaging conditions). In the context of membrane diffusion, this means that FCS allows studying lipid diffusion in fluid membranes, as well as the diffusion of loosely-bound proteins hovering above the membrane. SPT, on the other hand, is ideal to study the motions of membrane-inserted proteins, especially those presenting different conformations, but only allows studying lipid diffusion in relatively viscous membranes, such as supported lipid bilayers and cell membranes. View Full-Text
Keywords: lipid; bilayer; membrane; diffusion; fluorescence; confocal microscopy; total internal reflection fluorescence microscopy (TIRF); FCS; SPT lipid; bilayer; membrane; diffusion; fluorescence; confocal microscopy; total internal reflection fluorescence microscopy (TIRF); FCS; SPT
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Rose, M.; Hirmiz, N.; Moran-Mirabal, J.M.; Fradin, C. Lipid Diffusion in Supported Lipid Bilayers: A Comparison between Line-Scanning Fluorescence Correlation Spectroscopy and Single-Particle Tracking. Membranes 2015, 5, 702-721.

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