Abstract
Wide-field imaging of neural activity at a cellular resolution is a current challenge in neuroscience. To address this issue, wide-field two-photon microscopy has been developed; however, the field size is limited by the objective size. Here, we develop a micro-opto-mechanical device that rotates within the post-objective space between the objective and brain tissue. Two-photon microscopy with this device enables sub-second sequential calcium imaging of left and right mouse sensory forelimb areas 6 mm apart. When imaging the rostral and caudal motor forelimb areas (RFA and CFA) 2 mm apart, we found high pairwise correlations in spontaneous activity between RFA and CFA neurons and between an RFA neuron and its putative axons in CFA. While mice performed a sound-triggered forelimb-movement task, the population activity between RFA and CFA covaried across trials, although the field-averaged activity was similar across trials. The micro-opto-mechanical device in the post-objective space provides a novel and flexible design to clarify the correlation structure between distant brain areas at subcellular and population levels.
Highlights
Wide-field imaging of neural activity at a cellular resolution is a current challenge in neuroscience
A potential way to address this issue in imaging of the mouse cerebral cortex is to expand the field of view (FOV) in two-photon laser scanning microscopy (TPLSM) because this microscopy technique allows detection of the activity of multiple neurons in all cortical layers and at a cellular resolution[3,4,5,6,7,8]
When the objective is redesigned, it is necessary to redesign the pre-objective optics, ensuring that a satisfactory cellular resolution is attained; for the purposes of neuronal imaging, the axial resolution should be finer than 10–15 μm, which is the diameter of the neuronal soma[11]
Summary
Wide-field imaging of neural activity at a cellular resolution is a current challenge in neuroscience. A potential way to address this issue in imaging of the mouse cerebral cortex is to expand the field of view (FOV) in two-photon laser scanning microscopy (TPLSM) because this microscopy technique allows detection of the activity of multiple neurons in all cortical layers and at a cellular resolution[3,4,5,6,7,8]. The device rapidly switches the FOV position, without movement of the objective or the sample Combining this device with TPLSM enables sequential imaging of multiple distant (more than 6 mm) areas of the mouse brain, which can be stitched together to form a large continuous image area (1.2 × 3.5 mm2) with a cellular resolution. We demonstrate the utility of the device by studying functional correlations between two distant motor cortical areas at the levels of single axonal boutons, single neurons, and the neuronal population
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