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Summary: In contrast to other techniques utilized in physiological studies, calcium imaging can visualize target neurons located deep in the brain. Here, we present a protocol for one-photon calcium imaging of dorsal and ventral CA1 neurons in head-fixed mice. We describe procedures for injecting GCaMP6f virus, implanting a gradient-index (GRIN) lens, and installing a baseplate for Inscopix microscope mounting.For complete details on the use and execution of this protocol, please refer to Yun et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.