Authors | Spaink, Herman P., Cui, Chao, Wiweger, Malgorzata I., Jansen, Hans J., Veneman, Wouter J., Marin-Juez, Ruben, de Sonneville, Jan, Ordas, Anita, Torraca, V., van der Ent, Wietske, Leenders, William P., Meijer, Annemarie H., Snaar-Jagalska, B. Ewa and Dirks, Ron P. |
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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. |
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