| Description |
Zebrafish (Danio rerio) have similar genetic structure to humans and share 70% of human genes. Zebrafish have high fecundity and are relatively inexpensive to maintain. Researchers model the complex nature of human embryonic development and identify potential treatments for human diseases by using zebrafish. In recent years, zebrafish have become important to biomedical research and are now being used for a variety of research projects. To assist in this research, the Zebrafish Embryo Genotyper (ZEG) was developed. It is the only automated device that collects genetic material rapidly from 72 hours postfertilization, zebrafish embryos while maintaining embryo wellbeing. The device consists of a microabrasion chip and a platform that generates harmonic oscillations to obtain genetic material for genotyping. Results show that ZEG samples have 90% sensitivity and survivability when genotyped with high-resolution melting analysis (HRMA). However, the current design appears to be inconsistent when used with traditional gel electrophoresis. Additionally, the manufacturing time for the chip is lengthy. Therefore, a raster engraving fabrication technique was investigated to address these issues. The objectives of this study were to determine the factors that determined the surface texture profile characteristics of a manufactured ZEG chip, and an optimized operation protocol maximize sensitivity and embryo survival. Results show that the surface texture profile was the largest contributor to the inconsistent results. A new manufacturing and operating protocol was developed using a design of experiments vi (DOE) approach. The new design achieved over 80% PCR amplification sensitivity with gel electrophoresis and over 95% survivability consistently. The new design also reduced the manufacturing time by 90%. In summary, raster engraving created more consistent surface texture profile and yields more consistent PCR amplification and fish survivability. |
