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

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

Author information

1
Department of Molecular Cell Biology, Institute of Biology, Leiden University, The Netherlands. h.p.spaink@biology.leidenuniv.nl

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

PMID:
23769806
DOI:
10.1016/j.ymeth.2013.06.002
[Indexed for MEDLINE]
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