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An alternative method for delivering exogenous material into developing zebrafish embryos

Authors

  • Vikram Kohli,

    Corresponding author
    1. 9107-116 St, Ultrafast Photonics and Nano-Optics Laboratory, Centre for Nanoelectronics, Nanophotonics & Nanoscale Systems, Department of Electrical and Computer Engineering, University of Alberta, Edmonton, Alberta, T6G 2V4, Canada; telephone: 780-492-0756; fax: 780-492-1811
    • 9107-116 St, Ultrafast Photonics and Nano-Optics Laboratory, Centre for Nanoelectronics, Nanophotonics & Nanoscale Systems, Department of Electrical and Computer Engineering, University of Alberta, Edmonton, Alberta, T6G 2V4, Canada; telephone: 780-492-0756; fax: 780-492-1811.
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  • Vanesa Robles,

    1. CCMAR, Center for Marine Sciences, University of Algarve, Faro, Portugal
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  • M. Leonor Cancela,

    1. CCMAR, Center for Marine Sciences, University of Algarve, Faro, Portugal
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  • Jason P. Acker,

    1. Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, Alberta, Canada
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  • Andrew J. Waskiewicz,

    1. Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada
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  • Abdulhakem Y. Elezzabi

    1. 9107-116 St, Ultrafast Photonics and Nano-Optics Laboratory, Centre for Nanoelectronics, Nanophotonics & Nanoscale Systems, Department of Electrical and Computer Engineering, University of Alberta, Edmonton, Alberta, T6G 2V4, Canada; telephone: 780-492-0756; fax: 780-492-1811
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Abstract

Non-invasive manipulation of multicellular systems is important for medical and biological research. The ability to introduce, remove, or modify molecules in the intracellular environment is pivotal to our understanding of cellular structure and function. Herein, we report on an alternative method for introducing foreign material into developing embryos using the application of femtosecond (fs) laser pulses. When intense fs laser pulses are focused to a sub-micron spot, transient pores are formed, providing a transport pathway for the delivery of exogenous material into embryonic cells. In this study, zebrafish embryos were used as a model system to demonstrate the non-invasiveness of this applied delivery tool. Utilizing optically induced transient pores chorionated and dechorionated zebrafish embryos were successfully loaded with a fluorescent reporter molecule (fluorescein isothiocyanate), Streptavidin-conjugated quantum dots or DNA (Simian-CMV-EGFP). Pore formation was independent of the targeted location, with both blastomere-yolk interface and blastomere pores competent for delivery. Long-term survival of laser manipulated embryos to pec-fin stage was 89% and 100% for dechorionated and chorionated embryos, respectively. To our knowledge, this is the first report of DNA delivery into zebrafish embryos utilizing fs laser pulses. Biotechnol. Bioeng. 2007;98: 1230–1241. © 2007 Wiley Periodicals, Inc.

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