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Open AccessArticle

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
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Academic Editor: Maikel Rheinstadter
Membranes 2015, 5(4), 702-721; https://doi.org/10.3390/membranes5040702
Received: 17 October 2015 / Accepted: 6 November 2015 / Published: 13 November 2015
(This article belongs to the Special Issue Membrane Structure and Dynamics)
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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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. https://doi.org/10.3390/membranes5040702

AMA Style

Rose M, Hirmiz N, Moran-Mirabal JM, Fradin C. Lipid Diffusion in Supported Lipid Bilayers: A Comparison between Line-Scanning Fluorescence Correlation Spectroscopy and Single-Particle Tracking. Membranes. 2015; 5(4):702-721. https://doi.org/10.3390/membranes5040702

Chicago/Turabian Style

Rose, Markus; Hirmiz, Nehad; Moran-Mirabal, Jose M.; Fradin, Cécile. 2015. "Lipid Diffusion in Supported Lipid Bilayers: A Comparison between Line-Scanning Fluorescence Correlation Spectroscopy and Single-Particle Tracking" Membranes 5, no. 4: 702-721. https://doi.org/10.3390/membranes5040702

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