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1.
Anal Bioanal Chem ; 408(2): 599-607, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26537925

RESUMO

We have developed microfluidic devices with pressure-driven injection for electrophoretic analysis of amino acids, peptides, and proteins. The novelty of our approach lies in the use of an externally actuated on-chip peristaltic pump and closely spaced pneumatic valves that allow well-defined, small-volume sample plugs to be injected and separated by microchip electrophoresis. We fabricated three-layer poly(dimethylsiloxane) (PDMS) microfluidic devices. The fluidic layer had injection and separation channels, and the control layer had an externally actuated on-chip peristaltic pump and four pneumatic valves around the T-intersection to carry out sample injection. An unpatterned PDMS membrane layer was sandwiched between the fluidic and control layers as the actuated component in pumps and valves. Devices with the same peristaltic pump design but different valve spacings (100, 200, 300, and 400 µm) from the injection intersection were fabricated using soft lithographic techniques. Devices were characterized through fluorescent imaging of captured plugs of a fluorescein-labeled amino acid mixture and through microchip electrophoresis separations. A suitable combination of peak height, separation efficiency, and analysis time was obtained with a peristaltic pump actuation rate of 50 ms, an injection time of 30 s, and a 200-µm valve spacing. We demonstrated the injection of samples in different solutions and were able to achieve a 2.4-fold improvement in peak height and a 2.8-fold increase in separation efficiency though sample stacking. A comparison of pressure-driven injection and electrokinetic injection with the same injection time and separation voltage showed a 3.9-fold increase in peak height in pressure-based injection with comparable separation efficiency. Finally, the microchip systems were used to separate biomarkers implicated in pre-term birth. Although these devices have initially been demonstrated as a stand-alone microfluidic separation tool, they have strong potential to be integrated within more complex systems.


Assuntos
Biomarcadores/análise , Eletroforese em Microchip/métodos , Eletroforese em Microchip/instrumentação , Desenho de Equipamento , Feminino , Ferritinas/análise , Humanos , Recém-Nascido , Dispositivos Lab-On-A-Chip , Masculino , Peptídeos/análise , Gravidez , Nascimento a Termo
2.
Anal Chem ; 85(24): 11809-16, 2013 Dec 17.
Artigo em Inglês | MEDLINE | ID: mdl-24274592

RESUMO

Horizontal flow profiles having uniform velocities (3-13% RSD) at fixed heights across 0.5, 2.0, and 5.6 mm widths, with magnitudes of ≤124 µm/s, can be sustained along a ∼25.0 mm path using redox-magnetohydrodynamics (MHD) microfluidic pumping in a small volume (14.3 mm wide × 27.0 mm long × 620 µm high) on a chip. Uniform velocity profiles are important in moving volume elements without shape distortion for assays and separations for lab-on-a-chip applications. Fluid movement resulting from the MHD force (FB = j × B) was monitored with video microscopy by tracking 10 µm, polystyrene latex beads mixed into the solution. The ionic current density, j, was generated in 0.095 M K3Fe(CN)6, 0.095 M K4Fe(CN)6, and 0.095 M KCl by applying a constant current across a 0.5, 2.0, or 5.6 mm gap between an anode-cathode pair of electrodes, consisting of one to four shorted parallel, coplanar gold microbands [each 25.0 mm × 98 µm × âˆ¼100 nm (thickness), and separated by 102 µm] fabricated on an insulated silicon substrate. By shorting the increasing numbers of microbands together, increasing currents (118, 180, 246, and 307 µA) could be applied without electrode damage, and the impact of ionic current density gradients on velocity profiles over the anodes and cathodes could also be investigated. The magnetic field, B, was produced with a 0.36 T NdFeB permanent magnet beneath the chip. Data analysis was performed using particle image velocimetry software. A vertical flow profile was also obtained in the middle of the 5.6 mm gap.

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