Automated Zebrafish Chorion Removal and Single Embryo Placement: Optimizing Throughput of Zebrafish Developmental Toxicity Screens Public Deposited

http://ir.library.oregonstate.edu/concern/undergraduate_thesis_or_projects/r494vm63t

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  • The potential of the developing zebrafish model for toxicology and drug discovery is limited by inefficient approaches to manipulating and chemically exposing zebrafish embryos—namely, manual placement of embryos into 96- or 384-well plates and exposure of embryos while still in the chorion, a barrier of poorly characterized permeability enclosing the developing embryo. We report the automated dechorionation of 1600 embryos at once at 4 h postfertilization (hpf) and placement of the dechorionated embryos into 96-well plates for exposure by 6 hpf. The process removed ≥95% of the embryos from their chorions with 2% embryo mortality by 24 hpf, and 2% of the embryos malformed at 120 hpf. The robotic embryo placement allocated 6-hpf embryos to 94.7% ± 4.2% of the wells in multiple 96-well trials. The rate of embryo mortality was 2.8% (43 of 1536) from robotic handling, the rate of missed wells was 1.2% (18 of 1536), and the frequency of multipicks was <0.1%. Embryo malformations observed at 24 hpf occurred nearly twice as frequently from robotic handling (16 of 864; 1.9%) as from manual pipetting (9 of 864; 1%). There was no statistical difference between the success of performing the embryo placement robotically or manually.
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  • description.provenance : Submitted by Mushfiqur Sarker (sarkermu@onid.orst.edu) on 2012-05-27T06:48:59Z No. of bitstreams: 1 JALA-MushfiqSarker.pdf: 2096893 bytes, checksum: b2646ba26caf0fc41a058b58e9ab94ac (MD5)
  • description.provenance : Approved for entry into archive by Laura Wilson(laura.wilson@oregonstate.edu) on 2012-05-29T20:06:30Z (GMT) No. of bitstreams: 1 JALA-MushfiqSarker.pdf: 2096893 bytes, checksum: b2646ba26caf0fc41a058b58e9ab94ac (MD5)
  • description.provenance : Made available in DSpace on 2012-05-29T20:06:30Z (GMT). No. of bitstreams: 1 JALA-MushfiqSarker.pdf: 2096893 bytes, checksum: b2646ba26caf0fc41a058b58e9ab94ac (MD5) Previous issue date: 2012-02

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