A smart imaging workflow for organ-specific screening in a cystic kidney zebrafish disease model

• Verfasst von: Pandey, Gunjan [VerfasserIn]
• Verfasst von: Westhoff, Jens [VerfasserIn]
• Verfasst von: Schaefer, Franz [VerfasserIn]
• Verfasst von: Gehrig, Jochen [VerfasserIn]
• Titel: A smart imaging workflow for organ-specific screening in a cystic kidney zebrafish disease model
• Verf.angabe: Gunjan Pandey, Jens H. Westhoff, Franz Schaefer and Jochen Gehrig
• E-Jahr: 2019
• Jahr: 14 March 2019
• Umfang: 16 S.
• Fussnoten: Gesehen am 06.03.2020
• Titel Quelle: Enthalten in: International journal of molecular sciences
• Ort Quelle: Basel : Molecular Diversity Preservation International, 2000
• Jahr Quelle: 2019
• Band/Heft Quelle: 20(2019,6) Artikel-Nummer 1290, 16 Seiten
• ISSN Quelle: 1422-0067
• ISSN Quelle: 1661-6596
• DOI: doi:10.3390/ijms20061290
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: automated imaging
• Sach-SW: high-content screening
• Sach-SW: image analysis
• Sach-SW: zebrafish
• K10plus-PPN: 1691850519

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.