PUBLICATION

A High-Content Larval Zebrafish Brain Imaging Method for Small Molecule Drug Discovery

Authors
Liu, H., Chen, S., Huang, K., Kim, J., Mo, H., Iovine, R., Gendre, J., Pascal, P., Li, Q., Sun, Y., Dong, Z., Arkin, M., Guo, S., Huang, B.
ID
ZDB-PUB-161013-1
Date
2016
Source
PLoS One   11: e0164645 (Journal)
Registered Authors
Dong, Zhiqiang, Guo, Su, Li, Qiang
Keywords
Larvae, Fluorescence imaging, Zebrafish, Neuroimaging, Imaging techniques, Neurons, Eyes, Library screening
MeSH Terms
  • Animals
  • Brain/diagnostic imaging*
  • Brain/drug effects*
  • Brain/growth & development
  • Dopaminergic Neurons/cytology
  • Dopaminergic Neurons/drug effects
  • Drug Evaluation, Preclinical/methods*
  • High-Throughput Screening Assays/methods*
  • Larva/drug effects
  • Larva/ultrastructure
  • Microscopy/methods
  • Neuroimaging/methods*
  • Optical Imaging/methods
  • Small Molecule Libraries/pharmacology
  • Zebrafish*/growth & development
PubMed
27732643 Full text @ PLoS One
Abstract
Drug discovery in whole-organisms such as zebrafish is a promising approach for identifying biologically-relevant lead compounds. However, high content imaging of zebrafish at cellular resolution is challenging due to the difficulty in orienting larvae en masse such that the cell type of interest is in clear view. We report the development of the multi-pose imaging method, which uses 96-well round bottom plates combined with a standard liquid handler to repose the larvae within each well multiple times, such that an image in a specific orientation can be acquired. We have validated this method in a chemo-genetic zebrafish model of dopaminergic neuron degeneration. For this purpose, we have developed an analysis pipeline that identifies the larval brain in each image and then quantifies neuronal health in CellProfiler. Our method achieves a SSMD* score of 6.96 (robust Z'-factor of 0.56) and is suitable for screening libraries up to 105 compounds in size.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping