Micro-coils were designed to selectively activate pyramidal neurons (PNs), avoiding axonal stimulation, and potentially overcoming the limitations of conventional electric stimulation[1][2]. Finite element simulations showed a significant impact of micro-coil design on neuron activation selectivity[3]. A V-shaped bend at the micro-coil tip-enhanced field gradient selectivity vertically, leading to spatially confined cortical activation. In previous in vivo experiments, extracellular electrodes were necessary for recording neuronal action potentials, potentially increasing brain tissue damage[4]. Two-photon microscopy (2PM) with high-speed imaging allows noninvasive optical biopsy, facilitating the observation of activated neurons in diverse model organisms[5][6].However, combining 2PM with micro-coils for in vivo neurostimulation poses several challenges: e.g., ensuring low invasiveness, preventing large shadows during 2PM imaging, the small 2PM working distance, and aligning micro-coil probes parallel to PNs in layer 2. Addressing these challenges, L-shaped vertical micro-coil probes were made (Fig 1a), requiring fewer steps than our 2023 study[4]. Its cross-section parallel to the brain surface is also significantly smaller than previous probes, thus minimizing shadow interference during 2PM imaging and reducing.The new coils, inserted into the visual cortex of aesthetized mice, successfully induced spatially restricted neuronal activity, observed through fluorescence changes near the coil[2][3][4].*Our data and paper have not been presented at a neuroscience conference before.Fig 1. (a) Illustration of 2PM and vertical micro-coil. Optical microscopy images of (b) the previous micro-coil tip[4], and (c) the finished probe tip in this work. (d) 2PM image in mouse in vivo. (e) The relative changes in fluorescence of neurons.