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Reconstruction of firing rate changes across neuronal populations by temporally deconvolved Ca2+ imaging

Nature Methods volume 3, pages 377–383 (2006)Cite this article

Abstract

Methods to record action potential (AP) firing in many individual neurons are essential to unravel the function of complex neuronal circuits in the brain. A promising approach is bolus loading of Ca2+ indicators combined with multiphoton microscopy. Currently, however, this technique lacks cell-type specificity, has low temporal resolution and cannot resolve complex temporal firing patterns. Here we present simple solutions to these problems. We identified neuron types by colocalizing Ca2+ signals of a red-fluorescing indicator with genetically encoded markers. We reconstructed firing rate changes from Ca2+ signals by temporal deconvolution. This technique is efficient, dramatically enhances temporal resolution, facilitates data interpretation and permits analysis of odor-response patterns across thousands of neurons in the zebrafish olfactory bulb. Hence, temporally deconvolved Ca2+ imaging (TDCa imaging) resolves limitations of current optical recording techniques and is likely to be widely applicable because of its simplicity, robustness and generic principle.

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Figure 1: Two-photon imaging of neuronal calcium signals in the zebrafish olfactory bulb.
Figure 2: Relationship between APs and Ca2+ signals.
Figure 3: Temporal deconvolution of two-photon Ca2+ signals.
Figure 4: Quantitative analysis of TDCa imaging.
Figure 5: Analysis of odor-evoked activity patterns in the zebrafish olfactory bulb.

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Acknowledgements

We thank W. Denk, T. Euler, J. Kerr, G. Laurent, H. Riecke, P.H. Seeburg and members of the Friedrich laboratory for support, helpful discussions, and/or comments on the manuscript. This work was supported by the Max Planck-Society, the Deutsche Forschungsgemeinschaft (DFG; SFB 488), and a fellowship from the Boehringer Ingelheim Fonds to E.Y.

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Authors and Affiliations

  1. Department of Biomedical Optics, Max Planck Institute for Medical Research, Jahnstr. 29, Heidelberg, 69120, Germany

    Emre Yaksi & Rainer W Friedrich

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  1. Emre Yaksi
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  2. Rainer W Friedrich
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Correspondence to Rainer W Friedrich.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Principle of firing rate reconstruction by temporal deconvolution. (PDF 145 kb)

Supplementary Fig. 2

Decay time constants and their influence on deconvolution. (PDF 273 kb)

Supplementary Fig. 3

Iterative smooting procedure. (PDF 111 kb)

Supplementary Fig. 4

Deconvolution parameter search results. (PDF 83 kb)

Supplementary Fig. 5

Comparison of temporal deconvolution to other methods. (PDF 156 kb)

Supplementary Methods (PDF 121 kb)

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Yaksi, E., Friedrich, R. Reconstruction of firing rate changes across neuronal populations by temporally deconvolved Ca2+ imaging. Nat Methods 3, 377–383 (2006). https://doi.org/10.1038/nmeth874

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