Limited view correction in low-optical-NA photoacoustic microscopy

Photoacoustic microscope (PAM) with a low-optical NA suffers from a limited view along the optical axis, due to the coherent cancellation of acoustic pressure waves after being excited with a smoothly focused beam. Using larger-NA (NA > 0.3) objectives can readily overcome the limited-view problem, while the consequences are the shallow working distance and time-consuming depth scanning for large-volume imaging. Instead, we report an off-axis oblique detection strategy that is compatible with a low-optical-NA PAM for turning up the optical-axis structures. Comprehensive photoacoustic modeling and ex vivo phantom and in vivo mouse brain imaging experiments are conducted to validate the efficacy of correcting the limited view. Proof-of-concept experiment results show that the visibility of optical-axis structures can be greatly enhanced by making the detection angle off the optical axis larger than 45°, strongly recommending that off-axis oblique detection is a simple and cost-effective alternative method to solve the limited-view problems in low-optical-NA PAMs.