Morphology-Preserving Holotomography: Quantitative Analysis of 3D Organoid Dynamics

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Main Authors: Oh, ChulMin, Cho, Jimin, Park, Juyeon, Lee, Hoyeon, Park, YongKeun
Format: Preprint
Published: 2025
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author Oh, ChulMin
Cho, Jimin
Park, Juyeon
Lee, Hoyeon
Park, YongKeun
author_facet Oh, ChulMin
Cho, Jimin
Park, Juyeon
Lee, Hoyeon
Park, YongKeun
contents 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. The results demonstrate consistent segmentation across diverse geometries and reveal coordinated epithelial-lumen remodeling, breakdown of morphometric homeostasis during collapse, 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 three dimensions.
format Preprint
id arxiv_https___arxiv_org_abs_2512_22486
institution arXiv
publishDate 2025
record_format arxiv
spellingShingle Morphology-Preserving Holotomography: Quantitative Analysis of 3D Organoid Dynamics
Oh, ChulMin
Cho, Jimin
Park, Juyeon
Lee, Hoyeon
Park, YongKeun
Medical Physics
Optics
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. The results demonstrate consistent segmentation across diverse geometries and reveal coordinated epithelial-lumen remodeling, breakdown of morphometric homeostasis during collapse, 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 three dimensions.
title Morphology-Preserving Holotomography: Quantitative Analysis of 3D Organoid Dynamics
topic Medical Physics
Optics
url https://arxiv.org/abs/2512.22486