Rapid High-Resolution Mosaic Acquisition for Photoacoustic Remote Sensing

Mechanical stages are routinely used to scan large expanses of biological specimens in photoacoustic imaging. This is primarily due to the limited field of view (FOV) provided by optical scanning. However, stage scanning becomes impractical at higher scanning speeds, or potentially unfeasible with heavier samples. Also, the slow scan-rate of the stages makes high resolution scanning a time-consuming process. Some clinical applications such as microsurgery require submicron resolution in a reflection-mode configuration necessitating a method that can acquire large field of views with a small raster scanning step size. In this study, we describe a method that combines mechanical stages with optical scanning for the rapid acquisition of high-resolution large FOVs. Optical scanning is used to acquire small frames in a two-dimensional grid formed by the mechanical stages. These frames are captured with specific overlap for effective image registration. Using a step size of 200 nm, we demonstrate mosaics of carbon fiber networks with FOVs of 0.8 × 0.8 mm² captured in under 70 s with 1.2 µm image resolution. Larger mosaics yielding an imaging area of 3 × 3 mm² are also shown. The method is validated by imaging a 1 × 1 mm² section of unstained histopathological human tissue.