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Please use this identifier to cite or link to this item: http://hdl.handle.net/1942/10328

Title: Measuring Diffusion of Lipid-like Probes in Artificial and Natural Membranes by Raster Image Correlation Spectroscopy (RICS): Use of a Commercial Laser-Scanning Microscope with Analog Detection
Authors: GIELEN, Ellen
VAN DE VEN, Martin
Gratton, Enrico
Digman, Michelle
RIGO, Jean-Michel
Hofkens, Johan
Engelborghs, Yves
Issue Date: 2009
Publisher: ACS Publications
Citation: LANGMUIR, 25(9). p. 5209-5218
Abstract: The heterogeneity in composition and interaction within the cellular membrane translates into a wide range of diffusion coefficients of its constituents. Therefore, several complementary microfluorimetric techniques such as fluorescence correlation spectroscopy (FCS), fluorescence recovery after photobleaching (FRAP) and single-particle tracking (SPT) have to be applied to explore the dynamics of membrane components. The recently introduced raster image correlation spectroscopy (RICS) offers a much wider dynamic range than each of these methods separately and allows for spatial mapping of the dynamic properties. RICS is implemented on a confocal laser-scanning microscope (CLSM), and the wide dynamic range is achieved by exploiting the inherent time information carried by the scanning laser beam in the generation of the confocal images. The original introduction of RICS used two-photon excitation and photon counting detection. However, most CLSM systems are based on one-photon excitation with analog detection. Here we report on the performance of such a commercial CLSM (Zeiss LSM 510 META) in the study of the diffusion of the fluorescent lipid analog 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indodicarbocyanine perchlorate (Dil-C-18(5)) both in giant unilamellar vesicles and in the plasma membrane of living oligodendrocytes, i.e., the myelin-producing cells of the central nervous system. It is shown that RICS on a commercial CLSM with analog detection allows for reliable results in the study of membrane diffusion by removal of unwanted correlations introduced by the analog detection system. The results obtained compare well with those collected by FRAP and FCS.
Notes: Reprint Address: Ameloot, M (reprint author), Hasselt Univ, Biomed Res Inst, Lab Cell Physiol, Agoralaan,Bldg C, B-3590 Diepenbeek, Belgium
URI: http://hdl.handle.net/1942/10328
Link to publication: 10.1021/la8040538
DOI: 10.1021/la8040538
ISI #: 000265528600053
ISSN: 0743-7463
Category: A1
Type: Journal Contribution
Validation: ecoom, 2010
Appears in Collections: Research publications

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