| Summary: | 碩士 === 國立中興大學 === 分子生物學研究所 === 95 === Microfluidic biochip has been extensively used in biomedical research in recent years. Micro-system technology integrated with microarray may improve the gene expression quality and the efficiency of nucleic acid hybridization. In this study, we utilize a droplet-based microfluidic system to increase the DNA microarray mixing efficiency. With this device, the slow diffusion kinetics of the biomolecules reaction is expedited. Microfluidics with mixing effect enhances the hybridization efficiency and reduces the hybridization time to 15 minutes compared with conventional flat glass microarray, which takes from 4h to over night for hybridization. The miniaturization also reduces the hybridization volumes to 5 μL. To profile gene expression patterns, human lung adenocarcinoma cell lines with different invasion capabilities were employed to analyze and screen the differential gene expression for metastasis-associated genes. 891 EST clones associated with adenocarcinoma cell were amplified by polymerase chain reactions and spotted onto a 16 mm x 23 mm glass support, which is integrated with PMMA microchannels fabricated by CO2 laser machine system. The microchannel is 100μm deep and 150 μm wide. The rapid nucleic acid hybridization time is reduced to 15 min with higher signal to noise ratio and detection limit is 0.103 μg/μL. The microfluidic system allows faster and easier gene expression profiling than conventional flat glass microarray system. In the future, the microfluidic system could provide potential applications in other cancer research.
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