All-optical in vivo photoacoustic tomography by adaptive multilayer acoustic backpropagation

Photoacoustic tomography (PAT) combines high optical contrast with deep acoustic penetration, making it valuable for biomedical imaging. However, all-optical systems often face challenges in measuring the acoustic wave-induced displacements on rough and dynamic tissues surfaces. We present an all-optical PAT system enabling fast and high-resolution volumetric imaging <i>in vivo</i>. By integrating holographic microscopy with a soft cover layer and coherent averaging, the system detects ultrasound-induced surface displacements over a 10 × 10 mm² area with 0.5 nm sensitivity in 1 s. A novel backpropagation algorithm reconstructs a depth-selective pressure image from two consecutive displacement maps. The coherent summation of these depth-selective pressure images enables the reconstruction of a 3D acoustic pressure image. Using adaptive multilayer backpropagation, we achieve imaging depths of up to 5 mm, with lateral and axial resolutions of 158 µm and 92 µm, respectively, demonstrated through <i>in vivo</i> imaging of mouse vasculature and chicken embryo vessels.