High-speed in toto 3D imaging with isotropic resolution by scanning light-field tomography

In toto imaging of large-scale transparent samples or cleared tissue is in high demand in broad biological applications such as oncology, neuroscience, and developmental biology to understand the functions and organizations of large-scale cells at organ level. However, traditional methods usually face resolution degradation due to the missing cone problem and spatially nonuniform optical aberrations. In addition, dense axial scanning is often required with a small depth of field (DOF) during high-resolution imaging. Here, we propose a method, termed scanning light-field tomography (SLiT), which could rapidly image large-scale samples of 6 x 6 x 6 mm3 in toto with near isotropic cellular resolution (x, z <^> 17 mu m, y <^> 10 mu m) within 30 s. Both the versatility and reliability of SLiT are validated by comparing the imaging performance with confocal microscopy on diverse samples. With high imaging speed and strong robustness to heterogeneous 3D aberrations, SLiT provides a competitive alternative for in toto imaging of large-scale transparent samples, providing new possibilities for exploring large-scale intercellular organizations and interactions across multiple organs. (c) 2024 Optica Publishing Group under the terms of the Optica Open Access Publishing Agreement