High-speed imaging of living specimen was performed using two-photon microscopy equipped with a spinning-disk scanning unit. Typically, a high-peak-power laser light source is needed to simultaneously induce two-photon excitation processes at several hundred focal points, generating the limitations of excitable fluorophores. Therefore, a high-peak-power neodymium-based 918-nm laser light source was used for intravital imaging of the most popular fluorophores, green fluorescent proteins. As a result, the proposed system obtained approximately 30 times brighter fluorescent signal than that obtained using a conventional mode-locked titanium:sapphire laser light source. Furthermore, the system visualized four-dimensional (xyz-t) calcium responses of pancreatic acinar cells agonist stimulations in the living G-CaMP7-expressing mouse with 60 million mu m(3) volume. (C) 2020 The Authors. Published by Elsevier Inc.