Robotic injection of zebrafish embryos for high-throughput screening in disease models
- Authors
- 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.
- ID
- ZDB-PUB-130710-4
- 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
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.