Robotic injection of zebrafish embryos for high-throughput screening in disease models

Methods. 2013 Aug 15;62(3):246-54. doi: 10.1016/j.ymeth.2013.06.002. Epub 2013 Jun 11.

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.

Keywords: Cancer; High-throughput screening; Infectious disease; Microinjection; Robotics; Zebrafish.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Benchmarking
  • Disease Models, Animal
  • Embryo, Nonmammalian / immunology
  • Embryo, Nonmammalian / microbiology
  • Embryo, Nonmammalian / ultrastructure
  • Gene Knockdown Techniques
  • High-Throughput Screening Assays / instrumentation
  • High-Throughput Screening Assays / methods*
  • Humans
  • Larva / genetics*
  • Larva / immunology
  • Larva / microbiology
  • Larva / ultrastructure
  • Microinjections / methods*
  • Microscopy, Fluorescence
  • Morpholinos / administration & dosage
  • Mycobacterium tuberculosis / immunology
  • Neoplasm Transplantation
  • Oligonucleotides, Antisense / administration & dosage
  • Robotics / methods*
  • Staphylococcus epidermidis / immunology
  • Tumor Cells, Cultured / transplantation
  • Zebrafish / genetics*
  • Zebrafish / immunology
  • Zebrafish / microbiology

Substances

  • Morpholinos
  • Oligonucleotides, Antisense