Automated 3D light-sheet screening with high spatiotemporal resolution reveals mitotic phenotypes

• DOI: 10.1242/jcs.245043
• Publication Date: 2020-04-15T14:00:58Z
• Authors: Björn Eismann; Teresa G. Krieger; Jürgen Beneke; Ruben Bulkescher; Lukas Adam; Holger Erfle; Carl Herrmann; Roland Eils; Christian Conrad
• Keywords: High-Content Screening

3D cell cultures enable the in vitro study of dynamic biological processes such as cycle, but their use high-throughput screens remains impractical with conventional fluorescent microscopy. Here, we present a screening workflow for automated evaluation mitotic phenotypes by light-sheet After sample preparation liquid handling robot, spheroids are imaged 24 h toto dual-view inverted selective plane illumination microscope (diSPIM) much improved signal-to-noise ratio, higher imaging speed, isotropic resolution and reduced light exposure compared to spinning disc confocal microscope. A dedicated high-content image processing pipeline implements convolutional neural network-based phenotype classification. We illustrate potential our approach using siRNA knockdown epigenetic modification 28 target genes assessing phenotypic role mitosis. By rendering microscopy operational applications, this enables gene characterization or drug candidate tissue-like culture models.