Visualizing tumor angiogenesis and boundary with polygon-scanning multiscale photoacoustic microscopy

Recently, we developed an integrated optical-resolution (OR) and acoustic-resolution (AR) PAM, which has multiscale imaging capability using different resolutions. However, limited by the scanning method, a tradeoff exists between the imaging speed and field of view, which impedes its wider applications. Here, we present an improved multiscale PAM which achieves high-speed wide-field imaging based on a homemade polygon scanner. Encoder trigger mode was proposed to avoid jittering of the polygon scanner during imaging. Distortions caused by polygon scanning were analyzed theoretically and compared with traditional types of distortions in optical-scanning PAM. Then a depth correction method was proposed and verified to compensate for the distortions. System characterization of OR-PAM and AR-PAM was performed prior to in vivo imaging. Blood reperfusion of an in vivo mouse ear was imaged continuously to demonstrate the feasibility of the multiscale PAM for high-speed imaging. Results showed that the maximum B-scan rate could be 14.65 Hz in a fixed range of 10 mm. Compared with our previous multiscale system, the imaging speed of the improved system was increased by a factor of 12.35. In vivo imaging of a subcutaneously inoculated B-16 melanoma of a mouse was performed. Results showed that the blood vasculature around the melanoma could be resolved and the melanoma could be visualized at a depth up to 1.6 mm using the multiscale PAM.