Fluorescence correlation spectroscopy in small cytosolic compartments depends critically on the diffusion model used.
Fluorescence correlation spectroscopy (FCS) is a powerful technique for measuring low concentrations of fluorescent molecules and their diffusion constants. In the standard case, fluorescence fluctuations are measured in an open detection volume defined by the confocal optics. However, if FCS measurements are carried out in cellular processes that confine the detection volume, the standard FCS model leads to erroneous results. In this paper, we derive a modified FCS model that takes into account the confinement of the detection volume. Using this model, we have carried out the first FCS measurements in dendrites of cultured neurons. We further derive, for the case of confined diffusion, the limits within which the standard two- and three-dimensional diffusion models give reliable results.
Gennerich
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Physiologisches Institut, Universität Göttingen, D 37073 Göttingen, Germany.
Schild
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D000818:Animals; D002462:Cell Membrane / Q000502:physiology / Q000648:ultrastructure; D002478:Cells, Cultured; D003600:Cytosol / Q000502:physiology / Q000648:ultrastructure; D003712:Dendrites / Q000648:ultrastructure*; D004058:Diffusion; D005456:Fluorescent Dyes; D008954:Models, Biological; D008962:Models, Theoretical; D009474:Neurons / Q000166:cytology* / Q000502:physiology* / Q000648:ultrastructure; D009830:Olfactory Bulb / Q000166:cytology / Q000502:physiology; D018408:Patch-Clamp Techniques; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods; D014982:Xenopus laevis
D005456:Fluorescent Dyes