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...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2012-01-01
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| Series: | PLoS ONE |
| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22238673/?tool=EBI |
| _version_ | 1818735357486694400 |
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| 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 |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-18T00:19:58Z |
| format | Article |
| id | doaj.art-cb453aee935a411d96f8838e1cb39404 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-18T00:19:58Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-cb453aee935a411d96f8838e1cb394042022-12-21T21:27:22ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0171e2991610.1371/journal.pone.0029916Automated reporter quantification in vivo: high-throughput screening method for reporter-based assays in zebrafish.Steven L WalkerJunko ArigaJonathan R MathiasVeena CoothankandaswamyXiayang XieMartin DistelReinhard W KösterMichael J ParsonsKapil N BhallaMeera T SaxenaJeff S MummReporter-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.https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22238673/?tool=EBI |
| spellingShingle | 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. PLoS ONE |
| title | 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_short | 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 |
| url | https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22238673/?tool=EBI |
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