title: Live-cell imaging of dendritic spines by STED microscopy.
abstract: Time lapse fluorescence imaging has become one of the most important approaches in neurobiological research. In particular, both confocal and two-photon microscopy have been used to study activity-dependent changes in synaptic morphology. However, the diffraction-limited resolution of light microscopy is often inadequate, forcing researchers to complement the live cell imaging strategy by EM. Here, we report on the first use of a far-field optical technique with subdiffraction resolution to noninvasively image activity-dependent morphological plasticity of dendritic spines. Specifically we show that time lapse stimulated emission depletion imaging of dendritic spines of YFP-positive hippocampal neurons in organotypic slices outperforms confocal microscopy in revealing important structural details. The technique substantially improves the quantification of morphological parameters, such as the neck width and the curvature of the heads of spines, which are thought to play critical roles for the function and plasticity of synaptic connections.
authors.affiliation: Department of Cellular and Systems Neurobiology, Max Planck Institute of Neurobiology, 82152 Martinsried, Germany. naegerl@neuro.mpg.de
authors.forename: U Valentin
authors.initials: UV
authors.lastname: Nägerl
authors.forename: Katrin I
authors.initials: KI
authors.lastname: Willig
authors.forename: Birka
authors.initials: B
authors.lastname: Hein
authors.forename: Stefan W
authors.initials: SW
authors.lastname: Hell
authors.forename: Tobias
authors.initials: T
authors.lastname: Bonhoeffer
mesh_terms: D000818:Animals; D049229:Dendritic Spines / Q000648:ultrastructure*; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence / Q000379:methods*; D009473:Neuronal Plasticity / Q000502:physiology; D055095:Optics and Photonics