Significance: Functional brain imaging experiments in awake animals require meticulous monitoring of animal behavior to screen for spontaneous behavioral events. Although these events occur naturally, they can alter cell signaling and hemodynamic activity in the brain and confound functional brain imaging measurements.
Aim: We developed a centralized, user-friendly, and stand-alone platform that includes an animal fixation frame, compact peripheral sensors, and a portable data acquisition system. The affordable, integrated platform can benefit imaging experiments by monitoring animal behavior for motion detection and alertness levels as complementary readouts for brain activity measurements.
Approach: A custom acquisition system was designed using a powerful, inexpensive microcomputer. We customized an accelerometer and miniature camera modules for efficient, real-time monitoring of animal motion detection and pupil diameter. We then tested and validated the platform's performance with optical intrinsic signal imaging and GCaMP fluorescence calcium imaging in functional activation experiments in awake mice.
Results: The integrated platform shows promise for detecting spontaneous motion and pupil dilation while imaging. Stimulus-induced pupil dilation was found to initiate earlier than cortical hemodynamics with a slower rise time. Compared with neuronal calcium response, stimulus-induced pupil dilation initiated later with a slower rise time.
Conclusions: We developed an integrated platform to monitor animal motion and pupil dynamics. The device can be easily coupled and synchronized with optical brain imaging systems to monitor behavior, alertness, and spontaneous motion for awake animal studies.
Keywords: awake mouse imaging; calcium imaging; functional activation; optical intrinsic signal; pupillometry; spontaneous motion.
© 2025 The Authors.