Diffusion of myelin oligodendrocyte glycoprotein in living OLN-93 cells investigated by raster-scanning image correlation spectroscopy (RICS).
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.
Gielen
Ellen
E
Laboratory for Biomolecular Dynamics, Catholic University Leuven, Celestijnenlaan 200G, 3001, Heverlee, Belgium.
Smisdom
Nick
N
De Clercq
Ben
B
Vandeven
Martin
M
Gijsbers
Rik
R
Debyser
Zeger
Z
Rigo
Jean-Michel
JM
Hofkens
Johan
J
Engelborghs
Yves
Y
Ameloot
Marcel
M
D000818:Animals; D045744:Cell Line, Tumor; D003911:Dextrans / Q000378:metabolism; D003952:Diagnostic Imaging / Q000379:methods; D004058:Diffusion; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D021962:Membrane Microdomains / Q000378:metabolism; D009185:Myelin Proteins; D018992:Myelin-Associated Glycoprotein / Q000378:metabolism*; D063308:Myelin-Oligodendrocyte Glycoprotein; D009836:Oligodendroglia / Q000378:metabolism* / Q000502:physiology; D009837:Oligodendroglioma / Q000473:pathology*; D051381:Rats; D013057:Spectrum Analysis / Q000379:methods*
D003911:Dextrans; D005456:Fluorescent Dyes; C571328:Mog protein, rat; D009185:Myelin Proteins; D018992:Myelin-Associated Glycoprotein; D063308:Myelin-Oligodendrocyte Glycoprotein; D016650:Fluorescein-5-isothiocyanate