Related Experiment Video
Updated: Dec 24, 2025
08:37
Setting Up a Simple Light Sheet Microscope for In Toto Imaging of C. elegans Development
Published on: May 5, 2014
23.6K
Automated 3D light-sheet screening with high spatiotemporal resolution reveals mitotic phenotypes
Björn Eismann1,2, Teresa G Krieger1,2,3, Jürgen Beneke2,4
1Division of Theoretical Bioinformatics, German Cancer Research Center (DKFZ), Heidelberg 69120, Germany.
Journal of Cell Science
|April 17, 2020
Summary
This study introduces a new workflow using light-sheet microscopy for high-throughput screening of 3D cell cultures. It enables automated evaluation of cell division phenotypes, advancing drug discovery and gene characterization.
Area of Science:
- Cell Biology
- Microscopy
- Bioimaging
Background:
- 3D cell cultures offer advanced in vitro models for studying dynamic biological processes like the cell cycle.
- Conventional fluorescent microscopy limits the practical application of 3D cultures in high-throughput screening.
- Automated evaluation of cellular phenotypes in 3D models is crucial for biological discovery.
Purpose of the Study:
- To develop and validate a high-throughput screening workflow for automated evaluation of mitotic phenotypes in 3D cell cultures.
- To demonstrate the utility of light-sheet microscopy for imaging 3D cell cultures in a screening context.
- To enable efficient characterization of gene function and drug candidate screening in tissue-like models.
Main Methods:
- A workflow combining liquid handling robotics, dual-view inverted selective plane illumination microscopy (diSPIM), and a convolutional neural network-based image processing pipeline was developed.
- diSPIM was employed for rapid, high-resolution, in toto imaging of 3D cell cultures over 24 hours.
- siRNA knockdown and epigenetic modification of 28 mitotic genes were used to assess phenotypic roles in mitosis.
Main Results:
- The diSPIM system demonstrated superior signal-to-noise ratio, imaging speed, isotropic resolution, and reduced phototoxicity compared to spinning disc confocal microscopy.
- The image processing pipeline successfully classified mitotic phenotypes.
- The workflow effectively illustrated the phenotypic roles of targeted mitotic genes in 3D cell culture models.
Conclusions:
- Light-sheet microscopy can be effectively implemented for high-throughput screening applications with 3D cell cultures.
- This automated workflow facilitates efficient target gene characterization and drug candidate evaluation in physiologically relevant 3D models.
- The developed system significantly enhances the utility of 3D cell cultures for large-scale biological screening.
