title: A High-Content Larval Zebrafish Brain Imaging Method for Small Molecule Drug Discovery.
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.
authors.affiliation: Joint Graduate Program in Bioengineering, University of California, San Francisco and University of California, Berkeley, California, United States of America.
authors.forename: Harrison
authors.initials: H
authors.lastname: Liu
authors.affiliation: Small Molecule Discovery Center, University of California, San Francisco, San Francisco, California, United States of America.|Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, United States of America.
authors.forename: Steven
authors.initials: S
authors.lastname: Chen
authors.affiliation: Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, California, United States of America.
authors.forename: Kevin
authors.initials: K
authors.lastname: Huang
authors.affiliation: Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, California, United States of America.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Kim
authors.affiliation: Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, California, United States of America.
authors.forename: Han
authors.initials: H
authors.lastname: Mo
authors.affiliation: Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, California, United States of America.
authors.forename: Raffael
authors.initials: R
authors.lastname: Iovine
authors.affiliation: Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, California, United States of America.
authors.forename: Julie
authors.initials: J
authors.lastname: Gendre
authors.affiliation: Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, California, United States of America.
authors.forename: Pauline
authors.initials: P
authors.lastname: Pascal
authors.affiliation: Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, California, United States of America.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Li
authors.affiliation: Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, California, United States of America.
authors.forename: Yaping
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, California, United States of America.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Dong
authors.affiliation: Small Molecule Discovery Center, University of California, San Francisco, San Francisco, California, United States of America.
authors.forename: Michelle
authors.initials: M
authors.lastname: Arkin
authors.affiliation: Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, California, United States of America.
authors.forename: Su
authors.initials: S
authors.lastname: Guo
authors.affiliation: Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, United States of America.|Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, California, United States of America.
authors.forename: Bo
authors.identifier: http://orcid.org/0000-0003-1704-4141
authors.initials: B
authors.lastname: Huang
mesh_terms: D000818:Animals; D001921:Brain / Q000000981:diagnostic imaging* / Q000187:drug effects* / Q000254:growth & development; D059290:Dopaminergic Neurons / Q000166:cytology / Q000187:drug effects; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods*; D007814:Larva / Q000187:drug effects / Q000648:ultrastructure; D008853:Microscopy / Q000379:methods; D059906:Neuroimaging / Q000379:methods*; D061848:Optical Imaging / Q000379:methods; D054852:Small Molecule Libraries / Q000494:pharmacology; D015027:Zebrafish / Q000254:growth & development
chemical_list: D054852:Small Molecule Libraries