Single cell resolution 3D imaging and segmentation within intact live tissues.

ABSTRACT: Epithelial cells form diverse structures, from squamous spherical organoids to densely packed pseudostratified folded tissues. Quantification of cellular properties in these contexts requires high-resolution deep imaging and computational techniques to achieve truthful three-dimensional (3D) structural features. Here, we describe a detailed step-by-step protocol for sample preparation, imaging and deep-learning-assisted cell segmentation to achieve accurate quantification of fluorescently labelled individual cells in 3D within a live tissue: the Drosophila wing disc. We share the "lessons learned" through troubleshooting 3D imaging, including considerations on the choice of microscopy modality and settings (objective, sample mounting) and available segmentation methods. In addition, we inc

SUBMITTER: Paci G 

PROVIDER: S-EPMC12408837 | biostudies-literature | 2025 Sep

REPOSITORIES: biostudies-literature