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Free keywords:
Two-photon microscopy, In vivo calcium imaging, Multicell bolus loading, Olfactory bulb, Genetically encoded calcium indicators, Small-molecule calcium indicators, TN-XXL, Fluorescence resonance energy transfer
Abstract:
Functional properties of neuronal circuits can be best studied in vivo in the living mammalian brain. The use
of optical methods, like two-photon calcium imaging, permits analyses of network function at single-cell
resolution. This chapter provides a step-by-step description of this technique. Using mouse olfactory bulb
as a model system, we compare the performance of genetically encoded calcium sensor TN-XXL and smallmolecule
calcium indicators; describe how to choose the right calcium indicator and how to load it into the
cells of interest; discuss the use of cell type-specific markers and, finally, illustrate the application of this
technique for high-resolution in vivo imaging of sensory-driven neuronal activity