Multibeam continuous axial scanning two-photon microscopy for in vivo volumetric imaging in mouse brain

Mitsutoshi Ataka; Kohei Otomo; Ryosuke Enoki; Hirokazu Ishii; Motosuke Tsutsumi; Yuichi Kozawa; Shunichi Sato; and Tomomi Nemoto

Multibeam continuous axial scanning two-photon microscopy for in vivo volumetric imaging in mouse brain

Author : Mitsutoshi Ataka, Kohei Otomo, Ryosuke Enoki, Hirokazu Ishii, Motosuke Tsutsumi, Yuichi Kozawa, Shunichi Sato, and Tomomi Nemoto Journa Name: BIOMEDICAL OPTICS EXPRESS Country : USA Volume: 15 issue: 2 Year: 2024 Views : 201

Abstract:

This study presents an alternative approach for two-photon volumetric imaging that combines multibeam lateral scanning with continuous axial scanning using a confocal spinning-disk scanner and an electrically focus tunable lens. Using this proposed system, the brain of a living mouse could be imaged at a penetration depth of over 450??m from the surface. In vivo volumetric Ca2+ imaging at a volume rate of 1.5?Hz within a depth range of 130–200??m, was segmented with an axial pitch of approximately 5-µm and revealed spontaneous activity of neurons with their 3D positions. This study offers a practical microscope design equipped with compact scanners, a simple control system, and readily adjustable imaging parameters, which is crucial for the widespread adoption of two-photon volumetric imaging.