Three-dimensional voltage imaging in live larval zebrafish brain using fully genetically encoded voltage indicator

Voltage imaging has emerged as a powerful tool for recording membrane potential changes in living cells, offering a direct measurement of rapid neuronal events with high temporal precision. Since the brain is a three-dimensional circuit, it is essential to record signals across a volume. However, achieving effective three-dimensional voltage imaging over large neuronal populations remains challenging due to the need for high imaging speed, high signal-to-noise ratio, and extensive volume coverage. In this study, we demonstrate in vivo three-dimensional voltage imaging in larval zebrafish using oblique plane microscopy and QFDBD–QUAS–driven expression of the genetically encoded voltage indicator Ace-mNeon2-Kv2.1, achieving volumetric imaging rates of up to 200 volumes per second (VPS). This approach enables dye-free voltage imaging, simplifying experimental workflows and improving the reproducibility of in vivo voltage imaging experiments for investigating neuronal circuit dynamics in living zebrafish animal model.Datasets for paper titled " Three-dimensional voltage imaging in live larval zebrafish brain using fully genetically encoded voltage indicator" Experimental data: In vivo voltage imaging of larval zebrafish spinal cord File: OPM_Voltage_Zebrafish_6dpf_SpinalCord_Fig2.zip OPM_zebrafish_6dpf_SpinalCord_QUAS_AcemNeon2_1002Hz_raw.tif README.txt Note: This dataset corresponds to Figure 2 of the manuscript. It contains raw data before applying the SUPPORT denoising algorithm.