biostudies-literatureUnknown15(1)Nguyen VTPhotoacoustic microscopy (PAM) systems often face challenges in simultaneously achieving high speed, high resolution, high sensitivity, and a large field of view (FOV). To address this challenge, we have developed dual-channel PAM (DC-PAM) that can expand the FOV without compromising the imaging speed, detection sensitivity, or spatial resolution. DC-PAM has two identical, independent channels of laser excitation and acoustic detection. It exploits two facets of a single hexagon scanner to concurrently steer the dual excitation laser beams and the resultant acoustic waves. DC-PAM achieves an ultra-wide FOV of 22.5 × 24 mm² with a total functional imaging time of ~15 s. Proof-of-concept experiments were conducted using DC-PAM on freely-swimming zebrafish, hypoxia-challenged mice, and sleepiLight, science & applications91https://www.ebi.ac.uk/biostudies/studies/S-EPMC12852836biostudies-literatureDual-channel high-speed functional photoacoustic microscopy with ultra-wide field of view.PMC12852836Nguyen VTYao JLi JJayasundara NDelia JVu TCho SWDiSpirito ATaboada CMenozzi LfalseDual-channel high-speed functional photoacoustic microscopy with ultra-wide field of view.Photoacoustic microscopy (PAM) systems often face challenges in simultaneously achieving high speed, high resolution, high sensitivity, and a large field of view (FOV). To address this challenge, we have developed dual-channel PAM (DC-PAM) that can expand the FOV without compromising the imaging speed, detection sensitivity, or spatial resolution. DC-PAM has two identical, independent channels of laser excitation and acoustic detection. It exploits two facets of a single hexagon scanner to concurrently steer the dual excitation laser beams and the resultant acoustic waves. DC-PAM achieves an ultra-wide FOV of 22.5 × 24 mm² with a total functional imaging time of ~15 s. Proof-of-concept experiments were conducted using DC-PAM on freely-swimming zebrafish, hypoxia-challenged mice, and sleepi2026-01-01T00:00:00Z2026 Jan2026-06-17T07:00:41.136Z2026-06-17T03:10:34.812ZS-EPMC128528364160589410.1038/s41377-025-02114-3