Reconstruction of firing rate changes across neuronal populations by temporally deconvolved Ca2+ imaging.
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 Ca(2+) 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 Ca(2+) signals of a red-fluorescing indicator with genetically encoded markers. We reconstructed firing rate changes from Ca(2+) 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 Ca(2+) 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.
Yaksi
Emre
E
Max Planck Institute for Medical Research, Department of Biomedical Optics, Jahnstr. 29, 69120 Heidelberg, Germany.
Friedrich
Rainer W
RW
D000200:Action Potentials / Q000502:physiology*; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002118:Calcium / Q000032:analysis / Q000378:metabolism*; D020013:Calcium Signaling / Q000502:physiology*; D002413:Cations, Divalent; D004594:Electrophysiology / Q000379:methods*; D005456:Fluorescent Dyes / Q000032:analysis; D009474:Neurons / Q000502:physiology*; D009812:Odorants; D009830:Olfactory Bulb / Q000166:cytology / Q000502:physiology; D013050:Spectrometry, Fluorescence; D013194:Staining and Labeling; D013997:Time Factors; D015027:Zebrafish
D015415:Biomarkers; D002413:Cations, Divalent; D005456:Fluorescent Dyes; D002118:Calcium