A high-throughput imaging and nuclear segmentation analysis protocol for cleared 3D culture models.

ABSTRACT: Imaging and subsequent segmentation analysis in three-dimensional (3D) culture models are complicated by the light scattering that occurs when collecting fluorescent signal through multiple cell and extracellular matrix layers. For 3D cell culture models to be usable for drug discovery, effective and efficient imaging and analysis protocols need to be developed that enable high-throughput data acquisition and quantitative analysis of fluorescent signal. Here we report the first high-throughput protocol for optical clearing of spheroids, fluorescent high-content confocal imaging, 3D nuclear segmentation, and post-segmentation analysis. We demonstrate nuclear segmentation in multiple cell types, with accurate identification of fluorescently-labeled subpopulations, and develop a metric to ass

SUBMITTER: Boutin ME 

PROVIDER: S-EPMC6057966 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature