Automated reporter quantification in vivo: high-throughput screening method for reporter-based assays in zebrafish.
Reporter-based assays underlie many high-throughput screening (HTS) platforms, but most are limited to in vitro applications. Here, we report a simple whole-organism HTS method for quantifying changes in reporter intensity in individual zebrafish over time termed, Automated Reporter Quantification i...
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Public Library of Science (PLoS)
2012
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oai:doaj.org-article:cb453aee935a411d96f8838e1cb394042021-11-18T07:30:55ZAutomated reporter quantification in vivo: high-throughput screening method for reporter-based assays in zebrafish.1932-620310.1371/journal.pone.0029916https://doaj.org/article/cb453aee935a411d96f8838e1cb394042012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22238673/?tool=EBIhttps://doaj.org/toc/1932-6203Reporter-based assays underlie many high-throughput screening (HTS) platforms, but most are limited to in vitro applications. Here, we report a simple whole-organism HTS method for quantifying changes in reporter intensity in individual zebrafish over time termed, Automated Reporter Quantification in vivo (ARQiv). ARQiv differs from current "high-content" (e.g., confocal imaging-based) whole-organism screening technologies by providing a purely quantitative data acquisition approach that affords marked improvements in throughput. ARQiv uses a fluorescence microplate reader with specific detection functionalities necessary for robust quantification of reporter signals in vivo. This approach is: 1) Rapid; achieving true HTS capacities (i.e., >50,000 units per day), 2) Reproducible; attaining HTS-compatible assay quality (i.e., Z'-factors of ≥0.5), and 3) Flexible; amenable to nearly any reporter-based assay in zebrafish embryos, larvae, or juveniles. ARQiv is used here to quantify changes in: 1) Cell number; loss and regeneration of two different fluorescently tagged cell types (pancreatic beta cells and rod photoreceptors), 2) Cell signaling; relative activity of a transgenic Notch-signaling reporter, and 3) Cell metabolism; accumulation of reactive oxygen species. In summary, ARQiv is a versatile and readily accessible approach facilitating evaluation of genetic and/or chemical manipulations in living zebrafish that complements current "high-content" whole-organism screening methods by providing a first-tier in vivo HTS drug discovery platform.Steven L WalkerJunko ArigaJonathan R MathiasVeena CoothankandaswamyXiayang XieMartin DistelReinhard W KösterMichael J ParsonsKapil N BhallaMeera T SaxenaJeff S MummPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 1, p e29916 (2012) |
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EN |
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Medicine R Science Q Steven L Walker Junko Ariga Jonathan R Mathias Veena Coothankandaswamy Xiayang Xie Martin Distel Reinhard W Köster Michael J Parsons Kapil N Bhalla Meera T Saxena Jeff S Mumm Automated reporter quantification in vivo: high-throughput screening method for reporter-based assays in zebrafish. |
| description |
Reporter-based assays underlie many high-throughput screening (HTS) platforms, but most are limited to in vitro applications. Here, we report a simple whole-organism HTS method for quantifying changes in reporter intensity in individual zebrafish over time termed, Automated Reporter Quantification in vivo (ARQiv). ARQiv differs from current "high-content" (e.g., confocal imaging-based) whole-organism screening technologies by providing a purely quantitative data acquisition approach that affords marked improvements in throughput. ARQiv uses a fluorescence microplate reader with specific detection functionalities necessary for robust quantification of reporter signals in vivo. This approach is: 1) Rapid; achieving true HTS capacities (i.e., >50,000 units per day), 2) Reproducible; attaining HTS-compatible assay quality (i.e., Z'-factors of ≥0.5), and 3) Flexible; amenable to nearly any reporter-based assay in zebrafish embryos, larvae, or juveniles. ARQiv is used here to quantify changes in: 1) Cell number; loss and regeneration of two different fluorescently tagged cell types (pancreatic beta cells and rod photoreceptors), 2) Cell signaling; relative activity of a transgenic Notch-signaling reporter, and 3) Cell metabolism; accumulation of reactive oxygen species. In summary, ARQiv is a versatile and readily accessible approach facilitating evaluation of genetic and/or chemical manipulations in living zebrafish that complements current "high-content" whole-organism screening methods by providing a first-tier in vivo HTS drug discovery platform. |
| format |
article |
| author |
Steven L Walker Junko Ariga Jonathan R Mathias Veena Coothankandaswamy Xiayang Xie Martin Distel Reinhard W Köster Michael J Parsons Kapil N Bhalla Meera T Saxena Jeff S Mumm |
| author_facet |
Steven L Walker Junko Ariga Jonathan R Mathias Veena Coothankandaswamy Xiayang Xie Martin Distel Reinhard W Köster Michael J Parsons Kapil N Bhalla Meera T Saxena Jeff S Mumm |
| author_sort |
Steven L Walker |
| title |
Automated reporter quantification in vivo: high-throughput screening method for reporter-based assays in zebrafish. |
| title_short |
Automated reporter quantification in vivo: high-throughput screening method for reporter-based assays in zebrafish. |
| title_full |
Automated reporter quantification in vivo: high-throughput screening method for reporter-based assays in zebrafish. |
| title_fullStr |
Automated reporter quantification in vivo: high-throughput screening method for reporter-based assays in zebrafish. |
| title_full_unstemmed |
Automated reporter quantification in vivo: high-throughput screening method for reporter-based assays in zebrafish. |
| title_sort |
automated reporter quantification in vivo: high-throughput screening method for reporter-based assays in zebrafish. |
| publisher |
Public Library of Science (PLoS) |
| publishDate |
2012 |
| url |
https://doaj.org/article/cb453aee935a411d96f8838e1cb39404 |
| work_keys_str_mv |
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