@article{a374070a4cc04710a6bc8cfbb527a2e7,
title = "Robotic injection of zebrafish embryos for high-throughput screening in disease models",
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",
author = "Spaink, {Herman P.} and Chao Cui and Wiweger, {Malgorzata I.} and Jansen, {Hans J.} and Veneman, {Wouter J.} and Rub{\'e}n Mar{\'i}n-Juez and {De Sonneville}, Jan and Anita Ordas and Vincenzo Torraca and {van der Ent}, Wietske and Leenders, {William P.} and Meijer, {Annemarie H.} and Snaar-Jagalska, {B. Ewa} and Dirks, {Ron P.}",
note = "Funding Information: We thank Dr. A. Huttenlocher (University of Wisconsin) for antibodies and Dr. P. C.W. Hogendoorn (Leiden University Medical Centre) for the osteocarcoma cell lines. This research was supported by Project P5.03 IBIZA of the research program of the BioMedical Materials institute, co-funded by the Dutch Ministry of Economic Affairs and by the Smart Mix Program ( NWOA_6QY9BM ) of The Netherlands Ministry of Economic Affairs and of The Netherlands Ministry of Education, Culture and Science awarded to Leiden University and to ZF-screens BV. The authors further acknowledge financial support from the Leiden University Fund (LUF) for robotics and from Cyttron, in the Besluit Subsidies Investeringen Kennisinfrastructuur program, which in turn is financially supported by the Netherlands Organization for Scientific Research for imaging facilities. The COPAS system acquisition was in part supported by the Division for Earth and Life Sciences (ALW) with financial aid from the Netherlands Organization for Scientific Research ( NWO, 834.10.004 ). Additional support was obtained from the EU project ZF-Health ( FP7-Health-2009-242048 ), ZF-Cancer ( HEALTHF2-2008-201439 ) and FishForPharma ( PITN-GA-2011-289209 ). W. van der Ent was supported by a grant from the Dutch Children Cancer foundation (Kika).",
year = "2013",
month = aug,
day = "15",
doi = "10.1016/j.ymeth.2013.06.002",
language = "English",
volume = "62",
pages = "246--254",
journal = "Methods",
issn = "1046-2023",
publisher = "ACADEMIC PRESS INC",
number = "3",
}
