Abstract
Understanding brain function requires monitoring local and global brain dynamics. Two-photon imaging of the brain across mesoscopic scales has presented trade-offs between imaging area and acquisition speed. We describe a flexible cellular resolution two-photon microscope capable of simultaneous video rate acquisition of four independently targetable brain regions spanning an approximate five-millimeter field of view. With this system, we demonstrate the ability to measure calcium activity across mouse sensorimotor cortex at behaviorally relevant timescales.
Highlights
Understanding brain function requires monitoring local and global brain dynamics
The use of two independent scanning arms means that the imaging areas from each arm can be positioned anywhere within the total field of view (FOV) such that imaging areas from the first arm can overlap with imaging areas from the second arm
In summary, we demonstrate that the Quadroscope is a powerful new system to simultaneously capture neuronal dynamics across multiple cortical areas with high temporal resolution
Summary
Understanding brain function requires monitoring local and global brain dynamics. Twophoton imaging of the brain across mesoscopic scales has presented trade-offs between imaging area and acquisition speed. We describe a flexible cellular resolution two-photon microscope capable of simultaneous video rate acquisition of four independently targetable brain regions spanning an approximate five-millimeter field of view With this system, we demonstrate the ability to measure calcium activity across mouse sensorimotor cortex at behaviorally relevant timescales. The development of new two-photon microscopes has enabled imaging of increasingly large numbers of neurons across different brain areas This has been achieved through the design and fabrication of custom optical elements that support imaging over several millimeter fields of view while maintaining cellular resolution[2,3]. To address the gap in these technologies, we designed and constructed a quad-area large FOV two-photon microscope (Quadroscope) to achieve simultaneous, video-rate cellular-resolution imaging across four independently assignable FOVs. We demonstrate its utility in carrying out population imaging across mouse sensorimotor cortex during whisker-guided behavior at behaviorally relevant timescales
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