摘要：SummaryRecording the electrical activity of multiple neurons simultaneously would greatly facilitate studies on the function of neuronal circuits. The combination of the fast scanning by random-access multiphoton microscopy (RAMP) and the latest two-photon-compatible high-performance fluorescent genetically encoded voltage indicators (GEVIs) has enabled action potential detection in deep layers inin vivobrain. However, neuron connectivity analysis on optically recorded action potentials from multiple neurons in brain tissue has yet to be achieved. With high expression of a two-photon-compatible GEVI, ASAP3, viain uteroelectroporation and RAMP, we achieved voltage recording of spontaneous activities from multiple neurons in brain slice. We provide evidence for the developmental changes in intralaminar horizontal connections in somatosensory cortex layer 2/3 with a greater sensitivity than calcium imaging. This method thus enables investigation of neuronal network connectivity at the cellular resolution in brain tissue.Graphical AbstractDisplay OmittedHighlights•Kilohertz-order two-photon voltage imaging with single-trial acquisition•Readout spontaneous activity from multi-cells or subcellular regions simultaneously•Long-duration voltage imaging insuring functional studies of neuron circuit•Functional connections analysis with greater precision than calcium imagingOptical Imaging; Developmental Neuroscience; Cellular Neuroscience; Techniques in Neuroscience
