Optical-resolution parallel ultraviolet photoacoustic microscopy for slide-free histology | Science Advances

Intraoperative imaging of slide-free specimens is crucial for oncology surgeries, allowing surgeons to quickly identify tumor margins for precise surgical guidance. While high-resolution ultraviolet photoacoustic microscopy has been demonstrated for slide-free histology, the imaging speed is insufficient, due to the low laser repetition rate and the limited depth of field. To address these challenges, we present parallel ultraviolet photoacoustic microscopy (PUV-PAM) with simultaneous scanning of eight optical foci to acquire histology-like images of slide-free fresh specimens, improving the ultraviolet PAM imaging speed limited by low laser repetition rates. The PUV-PAM has achieved an imaging speed of 0.4 square millimeters per second (i.e., 4.2 minutes per square centimeter) at 1.3-micrometer resolution using a 50-kilohertz laser. In addition, we demonstrated the PUV-PAM with eight needle-shaped beams for an extended depth of field, allowing fast imaging of slide-free tissues with irregular surfaces. We believe that the PUV-PAM approach will enable rapid intraoperative photoacoustic histology and provide prospects for ultrafast optical-resolution PAM.