Development of Lab-on-a-chip Devices for Automated Zebrafish Embryo Bioassay
Published: 01-01-2011 In Publication
Abstract Zebrafish embryos have become one of the most popular model systems in biomedical and environmental research. However, current testing protocols using conventional multiwell plates rely heavily on time-consuming and labour-intensive manual handing. Static culture environments and low-throughput data collection are outdated with regards to meet the requirements of modern compound library screening. Herein, this research presents steps towards the development of a miniaturised and automated system for manipulating zebrafish embryos by using both innovative microfluidic lab-on-a-chip technologies and three-dimensional printing technologies. Four steps were taken to achieve this goal: (i) 3D printing technologies were explored to fabricate the lab-on-a-chip device. While 3D printing provided rapid manufacture of devices with high definition and optical transparency, as evidenced by SEM and confocal microscopy results, it caused significant toxicity in fish embryos after long-term exposure. (ii) The toxicity profile of a selection of 3D printing polymers was then extensively investigated using standard biotests. A chemical analysis was performed to reveal the compounds contributing to the toxicity. (iii) To avoid the use of toxic materials, a chip-based embryo trapping array was fabricated using biocompatible material PMMA. The chip allowed for automatic embryo loading, continuous reagent perfusion, and convenient image acquisition. The device was validated using both CFD simulations and biological experiments using reference toxicants. In addition, the embryo chip device was further developed to enable real-time metabolic level detection. (iv) A miniaturised and automated imaging platform, together with the high-throughput embryo trapping array and customised fluidic actuators, were prototyped.