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

The prevailing challenges in live unlabeled high-resolution imaging of native organoids stem from technical issues like complex sample handling and optical scattering in three-dimensional architectures. 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 successfully captured high-resolution morphological intricacies and dynamic events within mouse small intestinal organoids at a subcellular resolution. Furthermore, this method provides a unique advantage in differentiating between viable and non-viable organoids, thereby expanding its potential applications in organoid-based research. ### Competing Interest Statement M.J.L., J.H.L., G.K., H.J.K., H.H., S.M.L., and Y.K.P, have financial interests in Tomocube Inc., a company that commercializes holotomography and is one of the sponsors of the work.