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Time-lapse imaging of dendritic spines in vitro

Nature Protocols volume 1, pages 2399–2405 (2006)Cite this article

Abstract

Dendritic spines are small protrusions present postsynaptically at ∼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 (∼1 μm). 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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Figure 1: The imaging and electrophysiology station we currently use in our laboratory.
Figure 2: Motility plots.
Figure 3: Spine motility, depicted as changes in the shape factor over time.

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Authors and Affiliations

  1. Friedrich Miescher Institute, Maulbeerstrasse 66, Basel, 4058, Switzerland

    J Martin Verkuyl & Andrew Matus

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  1. J Martin Verkuyl
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  2. Andrew Matus
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Correspondence to Andrew Matus.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Video 1

Dendritic spine mobility (MOV 98 kb)

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Verkuyl, J., Matus, A. Time-lapse imaging of dendritic spines in vitro. Nat Protoc 1, 2399–2405 (2006). https://doi.org/10.1038/nprot.2006.357

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