A smart imaging workflow for organ-specific screening in a cystic kidney zebrafish disease model
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 develo...
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| Hauptverfasser: | , , , |
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| Dokumenttyp: | Article (Journal) |
| Sprache: | Englisch |
| Veröffentlicht: |
14 March 2019
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| In: |
International journal of molecular sciences
Year: 2019, Jahrgang: 20, Heft: 6 |
| ISSN: | 1422-0067 |
| DOI: | 10.3390/ijms20061290 |
| Online-Zugang: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.3390/ijms20061290 Verlag, lizenzpflichtig, Volltext: https://www.mdpi.com/1422-0067/20/6/1290 
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| Verfasserangaben: | Gunjan Pandey, Jens H. Westhoff, Franz Schaefer and Jochen Gehrig |
| Zusammenfassung: | 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. |
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| Beschreibung: | Gesehen am 06.03.2020 |
| Beschreibung: | Online Resource |
| ISSN: | 1422-0067 |
| DOI: | 10.3390/ijms20061290 |