Long-Term Three-Dimensional High-Resolution Imaging of Live Unlabeled Small Intestinal Organoids Using Low-Coherence Holotomography

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Abstract Organoids, which are miniature, in vitro versions of organs, present significant potential for studying human diseases and elucidating underlying mechanisms. To fully appreciate and understand the complex structure and dynamic biological phenomena of organoids, live imaging techniques play a crucial role in the field of organoid research. However, challenges in live, unlabeled high-resolution imaging of native organoids are prevalent, primarily due to the complexities of sample handling and optical scattering inherent in three-dimensional (3D) structures. Additionally, conventional imaging methods fall short in capturing the real-time dynamic processes of growing organoids. In this study, we introduce low-coherence holotomography as an advanced, label-free, quantitative imaging modality, designed to overcome related technical obstacles for long-term live imaging of 3D organoids. We demonstrate its efficacy by capturing high-resolution morphological details and dynamic activities within mouse small intestinal organoids at subcellular resolution. Moreover, our approach facilitates the distinction between viable and non-viable organoids, significantly enhancing its utility in organoid-based research. This advancement underscores the critical role of live imaging in organoid studies, offering a more comprehensive understanding of these complex systems.

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