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Diffusion of Myelin Oligodendrocyte Glycoprotein in Living OLN-93 Cells Investigated by Raster-Scanning Image Correlation Spectroscopy (RICS)

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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.

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Acknowledgements

We sincerely thank Prof. C. Richter-Landsberg for the generous gift of the OLN-93 oligodendroglial cells, Dr. W. Baron and Prof. D. Hoekstra for adapting the OLN-93 MOG-eGFP cell line, Prof. E. Gratton and Dr. M. Digman, Laboratory for Fluorescence Dynamics, University of Irvine, USA, for their help with RICS training and data analysis, Drs. K. Weisshart and M. Marx, Zeiss, Jena, Germany, for tracking scan parameter information, and Mrs. H. Penxten for performing viscosity measurements. This work was funded by the Research Council of the UHasselt, tUL, the K.U.Leuven (GOA/2006/02) and by a Ph.D grant of the Institute for the Promotion of Innovation through Science and Technology in Flanders (IWT-Vlaanderen). Support by the project IAP Functional Supramolecular Systems is gratefully acknowledged.

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Authors and Affiliations

  1. Laboratory for Biomolecular Dynamics, Catholic University Leuven, Celestijnenlaan 200G, 3001, Heverlee, Belgium

    Ellen Gielen & Yves Engelborghs

  2. Laboratory for Cell Physiology, Biomedical Research Institute, Hasselt University and transnationale Universiteit Limburg, Agoralaan, Bldg D, 3590, Diepenbeek, Belgium

    Ellen Gielen, Nick Smisdom, Ben De Clercq, Martin vandeVen, Jean-Michel Rigo & Marcel Ameloot

  3. Department of Applied Physics, Eindhoven University of Technology, Den Dolech 2, P.O. Box 513, 5600, MB, Eindhoven, The Netherlands

    Ben De Clercq

  4. Laboratory for Molecular Virology and Gene Therapy, Catholic University Leuven, Kapucijnenvoer 33 block i, 3000, Leuven, Belgium

    Rik Gijsbers & Zeger Debyser

  5. Laboratory of Photochemistry and Spectroscopy, Catholic University Leuven, Celestijnenlaan 200F, 3001, Heverlee, Belgium

    Johan Hofkens

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  1. Ellen Gielen
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  2. Nick Smisdom
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  3. Ben De Clercq
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  4. Martin vandeVen
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  10. Marcel Ameloot
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Corresponding author

Correspondence to Marcel Ameloot.

Additional information

Ellen Gielen and Nick Smisdom contributed equally to this work.

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Gielen, E., Smisdom, N., De Clercq, B. et al. Diffusion of Myelin Oligodendrocyte Glycoprotein in Living OLN-93 Cells Investigated by Raster-Scanning Image Correlation Spectroscopy (RICS). J Fluoresc 18, 813–819 (2008). https://doi.org/10.1007/s10895-007-0308-8

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  • DOI: https://doi.org/10.1007/s10895-007-0308-8

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