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Morphology-Preserving Holotomography: Quantitative Analysis of 3-D Organoid Dynamics
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DOI：10.1109/tmi.2026.3698497.png)
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Abstract
En 中文
Organoids are three-dimensional (3D) in vitro models for studying tissue development, disease progression, and physiological responses. Holotomography (HT) enables long-term, label-free imaging of live organoids by reconstructing volumetric refractive-index (RI) maps, but quantitative analysis is limited by the missing-cone artifact, which introduces anisotropic resolution and axial distortion. Here, we present a quantitative analysis framework that addresses the missing-cone problem at the level of image representation rather than reconstruction. We introduce morphology-preserving holotomography (MP-HT), a torus-shaped spatial filtering strategy that emphasizes high-spatial-frequency RI texture while suppressing low-frequency components most susceptible to missing-cone-induced distortion. Based on MP-HT, we develop a 3D segmentation pipeline for robust separation of epithelial and luminal structures, together with a model-based RI quantification approach that incorporates the system point spread function to enable morphology-independent estimation of dry-mass density and total dry mass. We apply the framework to long-term imaging of live hepatic organoids undergoing expansion, collapse, and fusion. In representative organoids, the framework provides consistent segmentation across diverse geometries and enables quantitative characterization of epithelial-lumen remodeling, collapse-associated loss of morphometric stability, and transient biophysical fluctuations during fusion. Overall, this work establishes a physically transparent and reproducible approach for quantitative, label-free analysis of organoid dynamics in 3D.
Keywords:
Holotomography
label-free imaging
organoid segmentation
quantitative spatiotemporal analysis
3D morphology
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