Affiliation:
1. National Institutes of Natural Sciences
2. Juntendo University
3. The Graduate School of Advanced Studies
4. Institute of Multidisciplinary Research for Advanced Materials, Tohoku University
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 μm from the surface. In vivo volumetric Ca2+ imaging at a volume rate of 1.5 Hz within a depth range of 130–200 μm, 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.
Funder
Exploratory Research Center on Life and Living Systems, National Institutes of Natural Sciences
Japan Agency for Medical Research and Development
Core Research for Evolutional Science and Technology
Japan Society for the Promotion of Science
Subject
Atomic and Molecular Physics, and Optics,Biotechnology