Long-term three-dimensional high-resolution imaging of live unlabeled small intestinal organoids via low-coherence holotomography

Organoids, which are miniature in vitro versions of organs, possess significant potential for studying human diseases and elucidating their underlying mechanisms. Live imaging techniques play a crucial role in organoid research and contribute to elucidating the complex structure and dynamic biological phenomena of organoids. However, live, unlabeled high-resolution imaging of native organoids is challenging, primarily owing to the complexities of sample handling and optical scattering inherent in three-dimensional (3D) structures. Additionally, conventional imaging methods fail to capture 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 several technical obstacles for long-term live imaging of 3D organoids. We demonstrate the efficacy of low-coherence holotomography 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 nonviable 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. Organoids, miniature 3D structures that imitate real organs, are grown in labs to study human biology and diseases. However, their complex structures and behaviors are hard to understand due to imaging technology limitations. In this study, researchers used a method called low-coherence holotomography to study mouse small intestinal organoids. This method let them observe organoids’ growth and drug responses in real-time, without altering their natural state. They conducted an experiment involving over 120 hours of continuous imaging, providing new insights into organoid development, cell dynamics, and drug responses. The study shows that low-coherence HT can reveal detailed 3D structures and changes within organoids, such as cell division and death, with high resolution. This research could revolutionize drug development and testing and provide new insights into human biology and diseases. This summary was initially drafted using artificial intelligence, then revised and fact-checked by the author.