PUBLICATION
A Smart Imaging Workflow for Organ-Specific Screening in a Cystic Kidney Zebrafish Disease Model
- Authors
- Pandey, G., Westhoff, J.H., Schaefer, F., Gehrig, J.
- ID
- ZDB-PUB-190317-1
- Date
- 2019
- Source
- International Journal of Molecular Sciences 20(6): (Journal)
- Registered Authors
- Gehrig, Jochen
- Keywords
- automated imaging, high-content screening, image analysis, zebrafish
- MeSH Terms
-
- Algorithms
- Animals
- Carrier Proteins/genetics*
- Disease Models, Animal
- Embryo, Nonmammalian/metabolism
- Gene Knockdown Techniques
- Humans
- Image Processing, Computer-Assisted/methods*
- Kidney/diagnostic imaging*
- Kidney/metabolism
- Organ Specificity
- Phenotype
- Polycystic Kidney Diseases/diagnostic imaging*
- Polycystic Kidney Diseases/metabolism
- Small Molecule Libraries/administration & dosage*
- Small Molecule Libraries/metabolism
- Software
- Workflow
- Zebrafish
- PubMed
- 30875791 Full text @ Int. J. Mol. Sci.
Citation
Pandey, G., Westhoff, J.H., Schaefer, F., Gehrig, J. (2019) A Smart Imaging Workflow for Organ-Specific Screening in a Cystic Kidney Zebrafish Disease Model. International Journal of Molecular Sciences. 20(6).
Abstract
The zebrafish is being increasingly used in biomedical research and drug discovery to conduct large-scale compound screening. However, there is a lack of accessible methodologies to enable automated imaging and scoring of tissue-specific phenotypes at enhanced resolution. Here, we present the development of an automated imaging pipeline to identify chemical modifiers of glomerular cyst formation in a zebrafish model for human cystic kidney disease. Morpholino-mediated knockdown of intraflagellar transport protein Ift172 in Tg(wt1b:EGFP) embryos was used to induce large glomerular cysts representing a robustly scorable phenotypic readout. Compound-treated embryos were consistently aligned within the cavities of agarose-filled microplates. By interfacing feature detection algorithms with automated microscopy, a smart imaging workflow for detection, centring and zooming in on regions of interests was established, which enabled the automated capturing of standardised higher resolution datasets of pronephric areas. High-content screening datasets were processed and analysed using custom-developed heuristic algorithms implemented in common open-source image analysis software. The workflow enables highly efficient profiling of entire compound libraries and scoring of kidney-specific morphological phenotypes in thousands of zebrafish embryos. The demonstrated toolset covers all the aspects of a complex whole organism screening assay and can be adapted to other organs, specimens or applications.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping