Live-cell imaging of dendritic spines by STED microscopy
Open accessAbstract
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.
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Neck diameters of spines located at 0-10 μm depth were originally measured to be on average ~40% reduced compared to confocal imaging.
“Neck diameters of spines located at 0–10 μm depth were originally measured to be on average ~40% reduced compared to confocal imaging (Nägerl et al., 2008),”
From Barriers in the brain: resolving dendritic spine morphology and compartmentalization (2014) · cites this paper as Nägerl et al., 2008
The first STED images of spines in organotypical slices expressing yellow fluorescent protein (YFP) in a sparse subset of neurons were published in 2008.
“The first STED images of spines in organotypical slices expressing yellow fluorescent protein (YFP) in a sparse subset of neurons were published in 2008 (Nägerl et al., 2008).”
From Barriers in the brain: resolving dendritic spine morphology and compartmentalization (2014) · cites this paper as Nägerl et al., 2008
Organotypic hippocampal slice cultures undergo key developmental steps similar to in vivo, including synapse maturation.
“Organotypic hippocampal slice cultures are known to undergo key developmental steps similar to those observed in vivo, including synapse maturation28–31, and have been previously used to study ramified microglia function32.”
From Microglia remodel synapses by presynaptic trogocytosis and spine head filopodia induction (2018) · cites this paper as [30]
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“We first discuss studies that examined the cytoplasmic coupling between spines and the dendritic shaft as a general measure of spine head isolation.”
Influential
- Authors
- Nägerl, U. V., Willig, K. I., Hein, B., Hell, S. W., & Bonhoeffer, T.
- Affiliations
- Max Planck Institute of Neurobiology (DE)
- Max Planck Institute for Biophysical Chemistry (DE)
- Venue
- Proceedings of the National Academy of Sciences
- Publisher
- National Academy of Sciences
- Published
- 2008-12-02
- Volume
- 105
- Issue
- 48
- Pages
- 18982-18987
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- PMC
- PMC2585941
- OpenAlex
- W1978431686
- Cited by
- 412
- References
- 24
- Citations by year
- 2026: 10 · 2025: 10 · 2024: 8 · 2023: 16 · 2022: 15 · 2021: 26 · 2020: 17 · 2019: 22 · 2018: 22 · 2017: 24 · 2016: 24 · 2015: 31 · 2014: 43 · 2013: 32 · 2012: 32
- FWCI
- 14.29
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Proceedings of the National Academy of Sciences (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Max Planck Institute for Plasma Physics (submittedVersion)
- Max Planck Institute for Plasma Physics (submittedVersion, other-oa)
- Max Planck Digital Library (submittedVersion, other-oa)
- GRO (submittedVersion)
- MPG.PuRe (Max Planck Society) (submittedVersion, other-oa)
- OPUS (Augsburg University) (submittedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Cell Image Analysis Techniques
- Keywords
- dendritic spines, STED microscopy, live-cell imaging, time-lapse fluorescence imaging, synaptic plasticity, hippocampal neurons, sub-diffraction resolution, dendritic spine morphology, spinal curvature, confocal microscopy comparison
- MeSH terms
- Animals, Image Processing, Computer-Assisted, Mice, Inbred C57BL, Mice, Transgenic, Microscopy, Fluorescence, Neuronal Plasticity, Dendritic Spines, Mice, Optics and Photonics
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