Automated manipulation of zebrafish embryos for high-throughput toxicology screening of nanomaterials Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/k0698b52n

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  • The field of nanotechnology has been rapidly developing new materials. These materials have become incorporated into consumer products and the environment after only minimal assessment of their effects on human health or the environment. The toxicity of these new nanomaterials remains unknown due to the unpredictable nature of the core material and its surface functionalization interactions. To understand the toxicity of nanomaterials, the embryonic zebrafish model is nearly ideal because it is a vertebrate model, is small size, external development, ability to produce numerous eggs, and has a completely sequenced genome. Due to the genomic similarity between humans and zebrafish, mechanistic studies are translatable to likely human effects. The largest bottleneck to implementing high-throughput nanomaterial toxicity testing is the tedious manipulation of the embryonic zebrafish in a systematic manner to obtain reliable results. This rate-limiting step provided an opportunity to utilize automated robotics for the highly repetitive tasks, which reduced errors, increased efficiency and made high-throughput screening practical at OSU. An automation system utilizing traditional macro-automation systems coupled to a microfluidic manipulator was used to move zebrafish embryos for high throughput screening of nanomaterial toxicity. In addition to moving embryos, the automated system was able to perform complex manipulations such as moving the embryo to a pulled glass needle to perform microinjection of antisense RNA to block the function of a specific gene. This system achieved a success rate as high as 99.5% when properly setup and calibrated. Thus, an efficient and rapid screening platform for nanomaterial toxicity was successfully created to establish the first vertebrate-based, high-throughput automation core at OSU.
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