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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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.
The authors declare no competing financial interests.
Principle of firing rate reconstruction by temporal deconvolution. (PDF 145 kb)
Decay time constants and their influence on deconvolution. (PDF 273 kb)
Iterative smooting procedure. (PDF 111 kb)
Deconvolution parameter search results. (PDF 83 kb)
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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