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Cellular layering is a hallmark of the mammalian neocortex. A key challenge in studying circuit function within the neocortex is to understand the spatial and temporal patterns of information flow between different columns and layers. In the paper, a mesoscopic functional imaging method for neural activities in mouse cortex in vivo was developed and demonstrated. We demonstrated the depth-resolved capability of 3D mesoscopic imaging by photostimulating the layer 5 neurons in Thy1-ChR2-YFP transgenic mice and further recorded the layer-specific functional projections between primary somatosensory cortex and primary motor cortex in vivo, following single whisker deflection.
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