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Simultaneous imaging of morphological plasticity and calcium dynamics in dendrites

Nature Protocols volume 1, pages 1859–1864 (2006)Cite this article

Abstract

The structure and function of the nervous system are intricately connected. To investigate their relationship it is essential to image neuronal structure and function simultaneously with high spatio-temporal resolution. For this purpose, we describe here a two-step strategy. First, to visualize neurons and their entire dendritic arborization in neuronal tissue, we use ballistic delivery or single-cell electroporation of a fluorescent calcium indicator and a red fluorescent dye. Second, dual wavelength wide-field fluorescence microscopy or confocal microscopy enables imaging structural plasticity of dendrites (including filopodia and spines) and calcium dynamics together. We routinely apply this strategy to developing neurons in live tissue, but mature neurons can also be loaded and imaged as described. For labeling cells and setting up imaging equipment, approximately 2 h are required.

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Figure 1: Single cell electroporation.
Figure 2: Setup for the consecutive imaging of two wavelengths using wide-field microscopy and a CCD camera.
Figure 3: Analysis of morphological plasticity and calcium dynamics.
Figure 4: Imaging structural plasticity and calcium dynamics in dendrites of a CA3 pyramidal neuron from the neonatal rat hippocampus.

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Acknowledgements

The Calistic technique was developed in collaboration with J. Demas, P. Kettunen, W.B. Gan and R.O.L. Wong.

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Authors and Affiliations

  1. Max-Planck Institute of Neurobiology, Am Klopferspitz 18, Planegg-Martinsried, 82152, Germany

    Susanne B Lang, Tobias Bonhoeffer & Christian Lohmann

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  1. Susanne B Lang
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  2. Tobias Bonhoeffer
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  3. Christian Lohmann
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Correspondence to Christian Lohmann.

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The authors declare no competing financial interests.

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Lang, S., Bonhoeffer, T. & Lohmann, C. Simultaneous imaging of morphological plasticity and calcium dynamics in dendrites. Nat Protoc 1, 1859–1864 (2006). https://doi.org/10.1038/nprot.2006.267

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