Optogenetic functional magnetic resonance imaging (ofMRI) is a powerful technique for investigating brain-wide dynamic responses to optogenetic modulation of specific cell populations. This protocol outlines a method for performing ofMRI in awake mice, integrating optogenetics with awake fMRI at 9.4 T using a high signal-to-noise ratio cryogenic coil. The detailed procedures include the preparation of flexible fiber optic implants, surgeries for opsin-encoding viral vector injection, fiber implantation, and head holder fixation, as well as habituation training for awake imaging, ofMRI scanning, and data analysis. The entire pipeline has been optimized to ensure high-quality data by enabling imaging within the limited space of the cryogenic coil and minimizing image distortion and head motion. In summary, ofMRI in awake mice enables precise local neural perturbation while providing whole-brain access, offering significant potential for advancing neuroscience research and preclinical imaging.