Live-cell imaging of dendritic spines by STED microscopy.
Proceedings of the National Academy of Sciences. 2008-11-21; 105(48): 18982-18987
DOI: 10.1073/pnas.0810028105
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1. Proc Natl Acad Sci U S A. 2008 Dec 2;105(48):18982-7. doi:
10.1073/pnas.0810028105. Epub 2008 Nov 21.
Live-cell imaging of dendritic spines by STED microscopy.
Nägerl UV(1), Willig KI, Hein B, Hell SW, Bonhoeffer T.
Author information:
(1)Department of Cellular and Systems Neurobiology, Max Planck Institute of
Neurobiology, 82152 Martinsried, Germany.
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.
DOI: 10.1073/pnas.0810028105
PMCID: PMC2585941
PMID: 19028874 [Indexed for MEDLINE]