We have developed a microchip for polymerase chain reaction (PCR) with polydimethylsiloxane (PDMS). PDMS has good characteristics: it is cheap, transparent, easy to fabricate and biocompatible. But in micro PCR, the porosity of PDMS causes several critical problems such as bubble formation, sample evaporation and protein adsorption. To solve those problems, we coated the micro PCR chips with Parylene film, which has low permeability to moisture and long-term stability. We investigated the influence of low thermal conductivity of PDMS and Parylene on the thermal characteristics of the PCR chips with numerical analysis. The thermal responses of micro PCR chips were compared for three materials: silicon, glass and PDMS. From the results, we identified appropriate thermal responses of the PDMS-based micro PCR chips by heating both the top and bottom sides. We could successfully amplify the angiotensin converting enzyme gene with as small a volume as 2 μl on the PDMS-based micro PCR chips without any additives.

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Stretching and folding, diffusion, and breakup are three basic processes that occur while mixing fluids. Although stretching and folding the interface of two fluids by rotation enables the mixing at microscale level in both low and high Reynolds number flows, rotation is not as effective at a low Reynolds number as at a high Reynolds number. Therefore, developing a rapid micromixer for microfluidic systems that can be used at a low Reynolds number is a challenging task, because it can demonstrate the full potential of microfluidic systems in commercial markets. Here, to enhance the mixing efficiency of a micromixer based on passive rotation, we present a breakup method. The breakup method not only generates interface actively but also enhances the diffusion process at the interface. With our novel design, over 70% mixing can be achieved only after passing through a 4 mm long microchannel. In this work, the mixer was easily fabricated with polydimethylsiloxane by soft lithography and a self-aligned bonding method with methanol. We analyzed the flow in the micromixer using the computational fluid dynamics method. Also, we conducted quantitative analyses using a confocal scanning microscope and image processing.

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Transfection of DNA molecules into mammalian cells with electric pulsations, which is so-called electroporation, is a powerful and widely used method that can be directly applied to gene therapy. However, very little is known about the basic mechanisms of DNA transfer and cell response to the electric pulse. We developed a microelectroporation chip with poly(dimethylsiloxane) (PDMS) to investigate the mechanism of electroporation as a first step of DNA transfer and to introduce the benefits of miniaturization into the genetic manipulation. The microelectroporation chip has a microchannel with a height of 20 microm and a length of 2 cm. Owing to the transparency of PDMS, we could in situ observe the uptake process of propidium iodide (PI) into SK-OV-3 cells, which shows promise in visualization of gene delivery in living cells. We also noticed the geometric effect on the degree of electroporation in microchannels with diverse channel width. This experimental result shows that the geometry can be another parameter to be considered for the electroporation when it is performed in microchannels with an exponential decaying pulse generator. Cell culturing is possible within the microelectroporation chip, and we also successfully transfected SK-OV-3 cells with enhanced green fluorescent protein genes, which demonstrates the feasibility of the microelectroporation chip in genetic manipulation.