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High-throughput in vivo vertebrate screening

Nature Methods volume 7, pages 634–636 (2010)Cite this article

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Abstract

We demonstrate a high-throughput platform for cellular-resolution in vivo chemical and genetic screens on zebrafish larvae. The system automatically loads zebrafish from reservoirs or multiwell plates, and positions and rotates them for high-speed confocal imaging and laser manipulation of both superficial and deep organs within 19 s without damage. We performed small-scale test screening of retinal axon guidance mutants and neuronal regeneration assays in combination with femtosecond laser microsurgery.

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Figure 1: Schematic of zebrafish manipulation and imaging platform.
Figure 2: Orientation, imaging and screening of zebrafish larvae.
Figure 3: Laser microsurgery and neuronal regeneration.
Figure 4: Quantitative assessment of zebrafish health.

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Acknowledgements

We thank the following funding sources: US National Institutes of Health Director's Innovator award (1-DP2-OD002989–01), Packard award in Science and Engineering, Alfred P. Sloan award, Sparc Grant from the Broad Institute, National Science Foundation Fellowship, Foxconn Sponsorship and the “La Caixa” Graduate Fellowship. We thank A. Amsterdam and N. Hopkins (MIT), and A. Schier and I. Woods (Harvard University) for the fluorescent lateral neuron line; C. Chien (University of Utah) for the fluorescent retinal ganglion line (robo2ti272z/ti272z with Tg(pou4f3:gap43-GFP)s356t)10,11; and S. Johnson (Washington University at St. Louis) and R. Jain and M. Granato (University of Pennsylvania) for the fluorescent Mauthner line Et(tol2:GFP) j1282b.

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Author notes

  1. Carlos Pardo-Martin and Tsung-Yao Chang: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology (MIT), Cambridge, Massachusetts, USA

    Carlos Pardo-Martin, Tsung-Yao Chang, Bryan Kyo Koo, Cody L Gilleland & Mehmet Fatih Yanik

  2. School of Engineering and Applied Sciences, Harvard University, Cambridge, Massachusetts, USA

    Carlos Pardo-Martin

  3. Division of Health Sciences and Technology, MIT, Cambridge, Massachusetts, USA

    Carlos Pardo-Martin

  4. Department of Biological Engineering, MIT, Cambridge, Massachusetts, USA

    Steven C Wasserman & Mehmet Fatih Yanik

Authors
  1. Carlos Pardo-Martin
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  2. Tsung-Yao Chang
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  3. Bryan Kyo Koo
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  4. Cody L Gilleland
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  5. Steven C Wasserman
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  6. Mehmet Fatih Yanik
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Contributions

C.P.-M., T.-Y.C., S.C.W. and M.F.Y. designed the experiments and wrote the manuscript. C.P.-M., T.-Y.C., B.K.K. and C.L.G. performed the experiments.

Corresponding author

Correspondence to Mehmet Fatih Yanik.

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Competing interests

M.F.Y. is founder and chief scientific advisor of Entera Pharmaceuticals. M.F.Y., C.L.G., Christopher B. Rohde and Matthew Angel have a pending patent on whole-organism screening methodologies (US patent application 12/713263) and M.F.Y., C.P.-M., T.-Y.C., C.L.G, and S.C.W. have a pending patent on zebrafish screening technology (US patent application 12/670882).

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Pardo-Martin, C., Chang, TY., Koo, B. et al. High-throughput in vivo vertebrate screening. Nat Methods 7, 634–636 (2010). https://doi.org/10.1038/nmeth.1481

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