Towards disposable lab-on-a-chip: Development of a low costly micro total analysis system (muTAS) for DNA analysis with poly(dimethylsiloxane) (PDMS).

摘要

The focus of this work is the development of a muTAS for DNA analysis, which allows us to perform DNA analysis with high throughput but low cost. The first goal of this research was to create a microfluidic device with a reasonable cost. In this work, a rapid protocol is demonstrated to create the micron-sized structures in the PDMS substrate material. This approach makes it possible to design, produce, and fabricate a microfluidic system with channel features >10 mum in PDMS in under 8 hours utilizing instrumentation common to most machine shops. The procedure involves the creation of a master template with negative features using high precision machining and the creation of an acrylic mold for casting channel structures into the PDMS substrate. The performance of the generated microfluidic system is evaluated by constructing a miniaturized capillary gel electrophoresis system for the analysis of DNA fragments with laser-induced fluorescence (LIF).; While "Lab-on-a-chip" technology promises to make biological and clinical analysis faster, cheaper, and easier than conventional technologies, a PDMS-glass hybrid miniaturized, integrated, and automated microfabricated chip-based fluidics system has been developed for DNA analysis. This system integrates PCR amplification and CGE analysis on chip. All steps for DNA analysis, including polymerase chain reaction (PCR) with an infrared radiation heating system, fluorescence dye labeling, transfer and injection of PCR product, sizing separation, and detection of PCR fragments, are integrated on a single chip. This work establishes the feasibility of performing all the steps of DNA analysis from real samples in a chip-based microfluidic integrated system, which represents the micro total analysis system concept.; Further integration of this work led to a more advanced muTAS device, which has integrated a couple more functions, pre-purification and pre-concentration, on-chip for detecting PCR fragments. The target DNA is identified based on its elution time. The analysis includes on-line PCR amplification utilizing IR-mediated thermocycling, hybridization of PCR fragments, pre-concentration of PCR fragments, and elution and detection on-chip. In summary, the goal of my research has been to develop a low cost and disposable PDMS muTAS device for DNA analysis, and to further the development of applications for muTAS.