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In the last 20 years, research activity using the zebrafish Danio rerio has increased dramatically, due in part to the ease of breeding and raising them, their genetic tractability, embryonic accessibility, and their imaging potential. In "Zebrafish: Methods and Protocols", expert researchers provide a collection of experimental techniques for the genetic modification, manipulation, and in vivo live imaging of zebrafish.
This volume collects sophisticated and practical techniques, including several not yet widely known, from laboratories around the world. Written in the highly successful "Methods in Molecular Biology" series format, chapters include brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, "Zebrafish: Methods and Protocols" seeks to aid and inspire scientists utilizing this versatile animal model in their research.
Part I. Mutagenesis and Mutants 1. Highly Efficient ENU Mutagenesis in Zebrafish Ewart de Bruijn, Edwin Cuppen, and Harma Feitsma 2. Production of Pseudotyped Retrovirus and the Generation of Proviral Transgenic Zebrafish Li-En Jao and Shawn M. Burgess 3. Undertaking a Successful Gynogenetic Haploid Screen in Zebrafish Judith E. Layton 4. Cryopreservation and in vitro Fertilization at the Zebrafish International Resource Center Carrie Carmichael, Monte Westerfield, and Zoltan M. Varga Part II. Transgenesis 5. Transient and Stable Transgenesis Using Tol2 Transposon Vectors Hiroshi Kikuta and Koichi Kawakami 6. Analysis of Genes and Genome by the Tol2-Mediated Gene and Enhancer Trap Methods Akihiro Urasaki and Koichi Kawakami 7. Bacterial Artificial Chromosome Transgenesis for Zebrafish Zhongan Yang, Hong Jiang, and Shuo Lin 8. Simple and Efficient Transgenesis with Meganuclease Constructs in Zebrafish Daniele Soroldoni, Benjamin M. Hogan, and Andrew C. Oates Part III. Tissue-Specific Manipulations 9. Nitroreductase-Mediated Cell Ablation in Transgenic Zebrafish Embryos 10. Focal Electroporation in Zebrafish Embryos and Larvae Marcel Tawk, Isaac H. Bianco, and Jonathan D. W. Clarke 11. Tissue Micromanipulation in Zebrafish Embryos Alexander Picker, Daniela Rollig, Olivier Pourquie, Andrew C. Oates, and Michael Brand Part IV. Analyzing Gene Expression 12. Zebrafish Spotted-Microarray for Genome-Wide Expression Profiling Experiments (Part I): Array Printing and Hybridization Siew Hong Lam, Sinnakarupan Mathavan, and Zhiyuan Gong 13. Zebrafish Spotted-Microarray for Genome-Wide Expression Profiling Experiments (Part II): Data Acquisition and Analysis Siew Hong Lam, R. Krishna Murthy Karuturi, and Zhiyuan Gong 14. Validating microRNA Target Transcripts Using Zebrafish Assays Luke Pase and Graham J. Lieschke Part V. Imaging 15. Multiple Embryo Time-Lapse Imaging of Zebrafish Development Leah Herrgen, Christian Schroter, Lola Bajard, and Andrew C. Oates 16. Live Cell Imaging of Zebrafish Leukocytes Chris Hall, Maria Vega Flores, Kathy Crosier, and Phil Crosier 17. Imaging Zebrafish Embryos by Two-Photon Excitation Time-Lapse Microscopy Lara Carvalho and Carl-Philipp Heisenberg 18. Morphological Analysis of the Zebrafish Digestive System Andrew J. Trotter, Adam C. Parslow, and Joan K. Heath 19. In toto Imaging of Embryogenesis with Confocal Time-Lapse Microscopy Sean G. Megason