AbstractExposure to time‐varying, low‐frequency and high‐intensity magnetic field (MF) induce electric field (EF) inside the human body, producing stimulus effects such as nerve fiber excitation or synaptic modulation. To measure such stimulus effects by low‐frequency MF exposure in real‐time, we developed a fluorescent recording system using optical fibers that is neither affected by the MF nor affects the MF distribution. In this study, a numerical calculation model composed of voxels with a 6.25 µm spatial resolution was developed. Using this numerical model, we evaluated the distribution of the EF generated inside three‐dimensional neuronal tissue called neural spheroid, under 50 Hz sinusoidal wave, 300 mT (root mean square) uniform MF exposure. We also investigated the influence of the optical fiber on the electric field distribution in neural spheroid. As a result, MF produced an induced EF in the neural spheroid of more than 4 V/m, well above the theoretical threshold of synaptic modulation. These results indicated that our experimental system was suitable for the evaluation of the threshold of stimulus effects using neural spheroid.