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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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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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)
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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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DOI: https://doi.org/10.1038/nmeth874
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