Summary
Although a common approach in large vertebrate embryos such as chick or frog, manipulation at the tissue level is only rarely applied to zebrafish embryos. Despite its relatively small size, the zebrafish embryo can be readily used for micromanipulations such as tissue and organ primordium transplantation, explantation, and microbead implantation, to study inductive tissue interactions and tissue autonomy of pleiotropic, mutant phenotypes or to isolate tissue for organotypic and primary cell culture or RNA isolation. Since this requires special handling techniques, tools, and tricks, which are rarely published and thus difficult to apply without hands-on demonstration, this article provides detailed instructions and protocols on tissue micromanipulation. The goal is to introduce a broader scientific audience to these surgical techniques, which can be applied to a wide range of questions and used as a starting point for many downstream applications in the genetically tractable zebrafish embryo.
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Acknowledgments
This work was supported by grants to MB from the Deutsche Forschungsgemeinschaft (SFB 655), and the EU (ZF-Models and Endotrack) and by the Max Planck Society. The authors would like to thank Mary-Lee Dequeant, Aurelie Krol, Alexander Aulehla, and Karen Echeverri for advice on PSM dissection and technical support, the fish facilities of the MPI-CBG and Biotechnology Center, Dresden and Stowers Institute, Kansas City, MO, USA, and Katrin Bergman (CRTD) for photography.
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Picker, A., Roellig, D., Pourquié, O., Oates, A.C., Brand, M. (2009). Tissue Micromanipulation in Zebrafish Embryos. In: Lieschke, G., Oates, A., Kawakami, K. (eds) Zebrafish. Methods in Molecular Biology, vol 546. Humana Press. https://doi.org/10.1007/978-1-60327-977-2_11
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DOI: https://doi.org/10.1007/978-1-60327-977-2_11
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