title: Robotic injection of zebrafish embryos for high-throughput screening in disease models.
abstract: The increasing use of zebrafish larvae for biomedical research applications is resulting in versatile models for a variety of human diseases. These models exploit the optical transparency of zebrafish larvae and the availability of a large genetic tool box. Here we present detailed protocols for the robotic injection of zebrafish embryos at very high accuracy with a speed of up to 2000 embryos per hour. These protocols are benchmarked for several applications: (1) the injection of DNA for obtaining transgenic animals, (2) the injection of antisense morpholinos that can be used for gene knock-down, (3) the injection of microbes for studying infectious disease, and (4) the injection of human cancer cells as a model for tumor progression. We show examples of how the injected embryos can be screened at high-throughput level using fluorescence analysis. Our methods open up new avenues for the use of zebrafish larvae for large compound screens in the search for new medicines.
authors.affiliation: Department of Molecular Cell Biology, Institute of Biology, Leiden University, The Netherlands. h.p.spaink@biology.leidenuniv.nl
authors.forename: Herman P
authors.initials: HP
authors.lastname: Spaink
authors.forename: Chao
authors.initials: C
authors.lastname: Cui
authors.forename: Malgorzata I
authors.initials: MI
authors.lastname: Wiweger
authors.forename: Hans J
authors.initials: HJ
authors.lastname: Jansen
authors.forename: Wouter J
authors.initials: WJ
authors.lastname: Veneman
authors.forename: Rubén
authors.initials: R
authors.lastname: Marín-Juez
authors.forename: Jan
authors.initials: J
authors.lastname: de Sonneville
authors.forename: Anita
authors.initials: A
authors.lastname: Ordas
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Torraca
authors.forename: Wietske
authors.initials: W
authors.lastname: van der Ent
authors.forename: William P
authors.initials: WP
authors.lastname: Leenders
authors.forename: Annemarie H
authors.initials: AH
authors.lastname: Meijer
authors.forename: B Ewa
authors.initials: BE
authors.lastname: Snaar-Jagalska
authors.forename: Ron P
authors.initials: RP
authors.lastname: Dirks
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D019985:Benchmarking; D004195:Disease Models, Animal; D004625:Embryo, Nonmammalian / Q000276:immunology / Q000382:microbiology / Q000648:ultrastructure; D055785:Gene Knockdown Techniques; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D007814:Larva / Q000235:genetics* / Q000276:immunology / Q000382:microbiology / Q000648:ultrastructure; D008845:Microinjections / Q000379:methods*; D008856:Microscopy, Fluorescence; D060172:Morpholinos / Q000008:administration & dosage; D009169:Mycobacterium tuberculosis / Q000276:immunology; D009368:Neoplasm Transplantation; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage; D012371:Robotics / Q000379:methods*; D013212:Staphylococcus epidermidis / Q000276:immunology; D014407:Tumor Cells, Cultured / Q000637:transplantation; D015027:Zebrafish / Q000235:genetics* / Q000276:immunology / Q000382:microbiology
chemical_list: D060172:Morpholinos; D016376:Oligonucleotides, Antisense
keywords: Cancer; High-throughput screening; Infectious disease; Microinjection; Robotics; Zebrafish