Cited by 13Neuroscience and Neuropharmacology ResearchAdvanced Fluorescence Microscopy TechniquesCell Image Analysis Techniques

Time-lapse imaging of dendritic spines in vitro

Verkuyl, J Martin, Matus, Andrew·2006·Nature Protocols

Abstract

Dendritic spines are small protrusions present postsynaptically at approximately 90% of excitatory synapses in the brain. Spines undergo rapid spontaneous changes in shape that are thought to be important for alterations in synaptic connectivity underlying learning and memory. Visualization of these dynamic changes in spine morphology are especially challenging because of the small size of spines (approximately 1 microm). Here we describe a microscope system, based on a spinning-disk confocal microscope, suitable for imaging mature dendritic spines in brain slice preparations, with a time resolution of seconds. We discuss two commonly used in vitro brain slice preparations and methods for transfecting them. Preparation and transfection require approximately 1 d, after which slices must be cultured for at least 21 d to obtain spines of mature morphology. We also describe imaging and computer analysis routines for studying spine motility. These procedures require in the order of 2 to 4 h.

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Crossref · 10.1038/nprot.2006.357

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  • 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.”

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Authors
Verkuyl, J. M., & Matus, A.
Affiliations
Friedrich Miescher Institute (CH)
Corresponding author
Matus, Andrew
Venue
Nature Protocols
Published
2006-12
Volume
1
Issue
5
Pages
2399-2405
ISSN
  • 1750-2799 (ISSN-L)
  • 1754-2189
OpenAlex
W1990142376
Cited by
13
References
31
Citations by year
2024: 1 · 2020: 1 · 2019: 3 · 2018: 2 · 2015: 1 · 2014: 3 · 2013: 1 · 2012: 1
FWCI
0.00
Type
journal article
Language
en
Open access
closed
Locations
  • Nature Protocols (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Cell Image Analysis Techniques
Keywords
dendritic spines, spine motility, dendritic spine morphology, time-lapse imaging, spinning disk confocal microscopy, brain slice preparation, in vitro transfection, excitatory synapses, synaptic plasticity, learning and memory
MeSH terms
Electrophysiology, Hippocampus, Time Factors, Microscopy, Confocal, Tissue Culture Techniques, Dendritic Spines

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