Acoustic-resolution-based photoacoustic microscopy with non-coaxial arrangements and a multiple vertical scan for high lateral resolution in-depth.

In conventional acoustic-resolution-based photoacoustic microscopy (ARPAM), a focused ultrasound transducer is placed coaxially with the laser beam to obtain the generated ultrasound signals. The information from deep regions can be greatly affected by the shallow targets. More importantly, in ARPAM the irreconcilable conflict between the lateral resolution and depth of fields has always been a major factor that lowers the imaging quality. In this work, an ARPAM system was developed, in which a non-coaxial arrangement of light illumination and acoustic detection was adopted to alleviate the influence of the tissue surface on the deep targets, and a focal zone integral algorithm was applied with a multiple scanning scheme to improve the lateral resolution. The system can achieve a consistent high lateral resolution of 0.5 mm over a large range in the axial direction. Both the phantom experiment and the chicken embryo in vivo results indicate that the proposed method can provide more in-depth information compared with the conventional ARPAM method. With the development of high repetition lasers and the advancement of image scanning technologies, the proposed method may play an important role in cerebral vascular imaging, superficial tumor imaging, and other related biomedical imaging applications.