期刊
METHODS
卷 46, 期 2, 页码 116-122出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2008.06.011
关键词
Fluorescence correlation spectroscopy; Single molecule; Model membrane; Giant unilamellar vesicle; Supported lipid bilayer; Lipid dynamics; Protein/lipid interactions; Membrane organization; Molecular interactions
资金
- Marie Curie Intra-European Fellowship
- 6th European Framework Program
Fluorescence correlation spectroscopy (FCS) is a powerful technique to study dynamic biomolecular processes. It allows the estimation of concentrations, diffusion coefficients, molecular interactions, and other processes causing fluctuations in the fluorescence intensity, thus yielding information about aggregation processes, enzymatic reactions, or partition coefficients. During the last years, FCS has been successfully applied to model and cellular membranes, proving to be a promising tool for the study of membrane dynamics and protein/lipid interactions. Here we describe the theoretical basis of FCS and some practical implications for its application in membrane studies. We discuss sources of potential artifacts, such as membrane undulations, positioning of the detection volume, and photobleaching. Special attention is paid to aspects related to instrumentation and sample preparation as well as data acquisition and analysis. Finally, we comment on some strategies recently developed for the specific improvement of FCS measurements on membranes. (C) 2008 Elsevier Inc. All rights reserved.
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