Diffusion of Myelin Oligodendrocyte Glycoprotein in Living OLN-93 Cells Investigated by Raster-Scanning Image Correlation Spectroscopy (RICS) |
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| Authors: | Ellen Gielen Nick Smisdom Ben De Clercq Martin vandeVen Rik Gijsbers Zeger Debyser Jean-Michel Rigo Johan Hofkens Yves Engelborghs Marcel Ameloot |
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| Affiliation: | (1) Laboratory for Biomolecular Dynamics, Catholic University Leuven, Celestijnenlaan 200G, 3001 Heverlee, Belgium;(2) Laboratory for Cell Physiology, Biomedical Research Institute, Hasselt University and transnationale Universiteit Limburg, Agoralaan, Bldg D, 3590 Diepenbeek, Belgium;(3) Department of Applied Physics, Eindhoven University of Technology, Den Dolech 2, P.O. Box 513, 5600, MB, Eindhoven, The Netherlands;(4) Laboratory for Molecular Virology and Gene Therapy, Catholic University Leuven, Kapucijnenvoer 33 block i, 3000 Leuven, Belgium;(5) Laboratory of Photochemistry and Spectroscopy, Catholic University Leuven, Celestijnenlaan 200F, 3001 Heverlee, Belgium |
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| Abstract: | Many membrane proteins and lipids are partially confined in substructures ranging from tens of nanometers to micrometers in size. Evidence for heterogeneities in the membrane of oligodendrocytes, i.e. the myelin-producing cells of the central nervous system, is almost exclusively based on detergent methods. However, as application of detergents can alter the membrane phase behaviour, it is important to investigate membrane heterogeneities in living cells. Here, we report on the first investigations of the diffusion behavior of the myelin-specific protein MOG (myelin oligodendrocyte glycoprotein) in OLN-93 as studied by the recently developed RICS (raster-scanning image correlation spectroscopy) technique. We implemented RICS on a standard confocal laser-scanning microscope with one-photon excitation and analog detection. Measurements on FITC-dextran were used to evaluate the performance of the system and the data analysis procedure. Ellen Gielen and Nick Smisdom contributed equally to this work. |
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| Keywords: | Raster-scanning image correlation spectroscopy Myelin oligodendrocyte glycoprotein OLN-93 Diffusion Lipid rafts |
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