Functional Magnetic Resonance Imaging (fMRI) with optogenetic neural manipulation is a powerful tool that enables brain-wide mapping of functional networks. To achieve flexible manipulation of neural excitation throughout the mouse cortex, we incorporated spatiotemporal programmable optogenetic stimuli generated by a digital micromirror device into an MR scanner via an optical fiber bundle. This approach offered versatility in space and time in planning the photostimulation pattern, combined with in situ optical imaging and cell-type or circuit-specific genetic targeting in individual mice. Brain-wide functional connectivity was successfully obtained by fMRI with optogenetic stimulation of atlas-based cortical regions. fMRI combined with flexible optogenetics opens a new path to investigate dynamic changes in functional brain states in the same animal.
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