Detachable head-mounted photoacoustic microscope in freely moving mice

Optical resolution photoacoustic microscopy (ORPAM) is a promising tool for investigating anatomical and functional dynamics in the cerebral cortex. However, observation in freely moving mice has been a longstanding challenge for ORPAM. In this Letter, we extended ORPAM from anesthetized, head-restrained to awake, freely moving mice by using a detachable head-mounted ORPAM probe. We used a micro-electro-mechanical-system scanner and a miniaturized piezoelectric ultrasonic detector to scan the excitation laser beam and detect generated photoacoustic signals, respectively. The probe weighs 1.8 g and has a large field of view of ∼3mm×3mm. We evaluated the performance of the probe by carrying out phantom experiments and the imaging of vascular networks in a mouse cerebral cortex. The results suggest that the ORPAM probe is capable of providing stable and high-quality ORPAM images in freely moving mice.