Gigavoxel-Scale Multiple-Scattering-Aware Lensless Holotomography

Abstract

Holotomography (HT) has revolutionized quantitative label-free 3D imaging, yet conventional lens-based implementations are fundamentally constrained in field-of-view (FOV) and imaging depth, limiting their utility for critical high-throughput applications in material and life sciences. Lensless HT (LHT) offers a promising alternative for large-volume examination, however existing approaches fail to accurately reconstruct highly scattering samples over extended depths, which remains a critical challenge in optical imaging field. Here, we introduce a gigavoxel-scale, multiple-scattering-aware LHT with a large FOV (surpassing 0.6 cm2), millimeter-scale axial range and pixel level (~2.4 micron) resolution. Our approach leverages a multi-wavelength, oblique-illumination hologram reconstruction and a robust, automatic illumination angle calibration, which are necessary for precise large-volume 3D holographic reconstruction. Moreover, we propose optimization-driven multi-slice tomographic framework to accurately capture multiple-scattering effects outperforming first order Born/Rytov-based inversions. To rigorously validate our method, we reconstruct bespoke multi-layer two-photon polymerized test structure over a 1.7 mm imaging depth and 25 mm2 FOV, yielding an unprecedented 3D space-bandwidth product exceeding a gigavoxel level. Furthermore, we demonstrate for the first time on-chip label-free imaging of entire 500-micron-thick tissue slice of optically-cleared mouse brain. With the proposed method, we aim to unlock powerful new capabilities for large-scale, quantitative, label-free 3D imaging across biomedicine, neuroscience, material sciences and beyond.