ZFIN ID: ZDB-PUB-120117-3
Automated reporter quantification in vivo: high-throughput screening method for reporter-based assays in zebrafish
Walker, S.L., Ariga, J., Mathias, J.R., Coothankandaswamy, V., Xie, X., Distel, M., Köster, R.W., Parsons, M.J., Bhalla, K.N., Saxena, M.T., and Mumm, J.S.
Date: 2012
Source: PLoS One 7(1): e29916 (Journal)
Registered Authors: Ariga, Junko, Köster, Reinhard W., Mathias, Jonathan, Mumm, Jeff, Parsons, Michael, Saxena, Meera T., Walker, Steven, Xie, Xiayang
Keywords: none
MeSH Terms:
  • Animals
  • Animals, Genetically Modified
  • Automation/methods
  • Bacterial Proteins/genetics
  • Bacterial Proteins/metabolism
  • Embryo, Nonmammalian
  • Gene Dosage*/physiology
  • Gene Expression Profiling/methods
  • Gene Expression Regulation, Developmental
  • Genes, Reporter*
  • High-Throughput Screening Assays/methods*
  • Image Processing, Computer-Assisted/methods*
  • Luminescent Proteins/genetics
  • Luminescent Proteins/metabolism
  • Osmolar Concentration
  • Reproducibility of Results
  • Validation Studies as Topic
  • Zebrafish*/embryology
  • Zebrafish*/genetics
PubMed: 22238673 Full text @ PLoS One
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ABSTRACT

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 e0.5), 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.

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