ZFIN ID: ZDB-PUB-130710-4
Robotic injection of zebrafish embryos for high-throughput screening in disease models
Spaink, H.P., Cui, C., Wiweger, M.I., Jansen, H.J., Veneman, W.J., Marín-Juez, R., de Sonneville, J., Ordas, A., Torraca, V., van der Ent, W., Leenders, W.P., Meijer, A.H., Snaar-Jagalska, B.E., and Dirks, R.P.
Date: 2013
Source: Methods (San Diego, Calif.) 62(3): 246-54 (Journal)
Registered Authors: de Sonneville, Jan, Marín-Juez, Rubén, Meijer, Annemarie H., Snaar-Jagalska, Ewa B., Spaink, Herman P., Torraca, Vincenzo, van der Ent, Wietske, Wiweger, Malgorzata
Keywords: zebrafish, microinjection, high-throughput screening, cancer, infectious disease, robotics
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
PubMed: 23769806 Full text @ Methods
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 compounds screens in the search for new medicines.

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