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Biomedical subjects

R L Chien

Publications and source records attributed to R L Chien.

5 recordsLinked to original sources

Sample stacking in laboratory-on-a-chip devices.

Sample stacking is a very important sample concentration technique. It has been used widely in capillary electrophoresis (CE). There are many different stacking techniques. One of the most popular techniques is called "field-amplified sample stacking" where an electric field discontinuity is set up across a concentration boundary. Charged analytes will then automatically stacked due to velocity changes after they cross the concentration boundary. There are several different strategies to perform sample stacking in microfluidic laboratory-on-a-chip devices. One could simply inject a plug of low concentration buffer containing sample into a channel surrounded by high concentration buffer. The electric field is then applied to stack the sample and move the whole plug into the separation channel. One could also stack the sample in a side channel adjacent to the separation channel. The disadvantage of this sample stacking technique is the difficulty in control of the precise location of stacked sample. We present a new sample stacking technique applied specifically to microfluidic laboratory-on-a-chip devices. Up to hundreds of fold increases in sample concentration can be achieved. We have also combined this stacking with electrophoretic separation in the same device.

Electrophoresis, Capillary↗

A fluorogenic assay using pressure-driven flow on a microchip.

A fluorogenic assay for human T-cell phosphatase (TCPTP) was conducted on an etched glass microchip using pressure driven flow. The TCPTP enzyme catalyzes the removal of a phosphate group from 6,8-difluoro-4-methylumbelliferyl/phosphate (DiFMUP) to produce the fluorogenic product 6,8-difluoro-4-methylumbelliferone (DiFMU). Enzyme assays with real-time on-chip dilution were performed in both low-viscosity (1 cP) buffer and an enzyme solution containing 50% glycerol (6 cP). Single side channels connect a series of reagent wells to a main channel where the fluorescent product of the enzyme reaction passes the detector region. Flow regulation of mixed viscosity fluids requires a pressure control on each arm of the chip contributing to the overall flow. An 8-channel pressure controller was built to regulate the air pressure above all wells feeding channels of the chip, thereby controlling the dilution ratios of buffer, substrate and enzyme. Well pressures maintained a constant concentration of enzyme in the detector channel while adjusting the flow contribution of substrate and buffer. The substrate concentration was stepped over two orders of magnitude while verifying fluid dilutions using marker dyes. The kinetic parameters, Km, Vmax, and Kcat, showed good agreement with the values determined using a standard well plate and fluorometer.

Chromogenic Compounds↗

Multiport flow-control system for lab-on-a-chip microfluidic devices.

A multiport system suitable for pressure control on a lab-on-a-chip microfluidic device is described. An algorithm and a strategy for calculating pressures were developed to control the flow from multiple reservoirs for the microfluidic devices. Dye mixing and enzyme assay titration experiments were performed using pressure-driven flow only. Results show a good linear response over two orders of dynamic range.

Algorithms↗