High-Performance DNA Separations in Microchip Electrophoresis Systems

Bousse, Luc; Dubrow, Bob; Ulfelder, Kathi J.

doi:10.1007/978-94-011-5286-0_64

Cited by 15 publications

(13 citation statements)

References 6 publications

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“…We use the resolution length (RSL) and theoretical plates per meter as readouts of the different techniques (see expression in the Method section) and report this data in plots (Fig. 7) as well as in Table 1. Micro-capillary electrophoresis (MCE) devices with polymer matrices like linear polyacrylamide can be used for DNA separation in the size range of 1-10 kbp (Bousse, Dubrow, and Ulfelder 1998;Mueller et al 2000;Lu and Liu 2006) with high resolution and theoretical plate numbers. An extension of the separation size range to 15 or 20 kbp can be obtained using low viscosity polymers (F. Xu et al 2003) and with the addition bacterial cellulose fibrils to the low viscosity sieving medium (Tabuchi and Baba 2005).…”

Section: Kbp Dna Band Broadening and Splitting Into Two Bands In Si/gmentioning

confidence: 99%

Single-step electrohydrodynamic separation of 1–150 kbp in less than 5 min using homogeneous glass/adhesive/glass microchips

Chami

Milon

Rojas

et al. 2020

Talanta

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Electrohydrodynamic migration, which is based on hydrodynamic actuation with an opposing electrophoretic force, enables the separation of DNA molecules of 3 to 100 kbp in capillary systems within one hour. Here, we wish to enhance these performances using microchip technologies. This study starts with the development of a fabrication process to obtain microchips with uniform surfaces, which is motivated by our observation that band splitting occurs in microchannels made out of heterogeneous materials such as glass and silicon. The resulting glass-adhesive-glass microchips feature the highest reported bonding strength of 11 MPa for such materials (115 kgf/cm 2), a high lateral resolution of critical dimension 5 µm, and minimal auto-fluorescence. These devices enable us to report the separation of 13 DNA bands in the size range of 1-150 kbp in one experiment of 5 minutes, i.e. 13 times faster than with capillary electrophoresis. In turn, we provide evidence that band splitting in heterogeneous glasssilicon microchips arises from differential Electro-Osmotic Flow (EOF) in between the upper and lower walls of the channel. We further indicate that differential EOF occurs in microchip electrophoresis and constitutes an underappreciated source of band broadening. We finally prove that our separation data compare favorably to state-of-the-art microchip technologies in terms of resolution length and theoretical plate numbers.

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Section: Kbp Dna Band Broadening and Splitting Into Two Bands In Si/gmentioning

confidence: 99%

Single-step electrohydrodynamic separation of 1–150 kbp in less than 5 min using homogeneous glass/adhesive/glass microchips

Chami

Milon

Rojas

et al. 2020

Talanta

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“…The channels are etched into fused-silica wafers and sealed by thermal bonding as previously described. 4 A 50 mm internal diameter fused-silica capillary is attached perpendicularly to the plane of the chip as shown in Figures 1 and 2. 5 To prepare the chip for analysis, wells 3, 5, 6, and 8 are filled with a proprietary polymer/dye solution that acts as the separation medium.…”

Section: Microfluidic Chip Designmentioning

confidence: 99%

“…Microfluidic devices hold great promise for automating many types of laboratory experimentation, including electrophoresis. [1][2][3][4] The first commercial instrument for microfluidic electrophoresis of nucleic acids and proteins, the Agilent 2100 Bioanalyzer, was introduced in 1999. This platform was co-developed by Agilent Technologies and Caliper Technologies and performs electrophoresis within microfluidic channels etched into glass wafers (termed "chips").…”

Section: Introductionmentioning

confidence: 99%

Automated Lab-on-a-Chip Analysis of DNA Fragments

Ausserer

Bousse

Gallagher

et al. 2001

JALA: Journal of the Association for Laboratory Automation

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T he design of a fully automated system for the analysis of DNA fragments in 96-well plates is described. Microfluidic technology is used to integrate sample loading, electrophoretic analysis, and fragment detection onto a miniature lab-on-a-chip device. The microfluidic chip operates in an instrument platform that automates sample access, data collection, and data reporting. Each microfluidic chip provides sizing and concentration values for more than 1000 DNA samples.

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“…Such intersections are surprisingly powerful tools that enable the definition of sample plugs at the picoliter level [2]; this in turn allows microfabricated electrokinetic systems to outperform their conventional counterparts by orders of magnitude [3]. The switching components are employed in separation and dispensing systems to inject the sample from the load channel to the separation channel.…”

Section: Introductionmentioning

confidence: 99%

Optimization of sample injection components in electrokinetic microfluidic systems

Bousse¹,

Minalla²,

Deshpande³

et al. 1999

Technical Digest. IEEE International MEMS 99 Conference. Twelfth IEEE International Conference on Micro Electro Mechanical Syst

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This paper presents experimental data, simulation tools (FlumeCAD), simulation results, and their use together to analyze and improve the designs of electrokinetic injection and switching components for microchemical fluidic systems.INTRODUCTION Electrokinetic microfluidic microsystems are powerful analytical tools for many applications, such as nucleic acid analysis, enzyme assays, and immunoassays [ 1-61. Such systems have gained considerable importance as components in micron-scale integrated chemicaybiochemical analysis or synthesis systems, also referred to as lab-on-a-chip. The basic "unit process" operations in these systems are sample injection, mixing, chemical reaction or modification, separation, and detection. Assembling a system of many "unit process" nodes requires one or more transport mechanisms to move sample and reagents through the "wires" of the system. Many of these systems rely on electrokinetic physics as their transport mechanism, although pressure and pneumatic applications have also been demonstrated. Complicated relationships exist between the microchannel geometries, the conditions under which the devices operate, and the behavior of the multicomponent fluids transported in these channels. In the past researchers have been forced to use costly trial and error methods to understand and design such microfluidic systems.CAD tools can be a valuable aid in the design of microfluidic systems. Numerical analyses provide significant insight into the fluid mechanics in these systems. They allow the extraction of material and flow properties that are generally not well documented, or that vary from application to application or from one manufacturing technology to another. Furthermore such tools help the designer to explore a much larger space of designs than is easily available from experiment, and do so in a quantitative way which enables the extraction of key parameters for improved or optimal operation of common microchemical system components.In this paper we include experimental data from some electrokinetic injection and switching components, as well as matching simulations of those components.We then demonstrate the use of the simulation tools to generate virtual experiments to help the designer choose good or optimal settings for the pinch field during injection and good or optimal settings for the switching field in a switch component.The simplest switching component is an intersection of two channels. Such intersections are surprisingly powerful tools that enable the definition of sample plugs at the picoliter level [2]; this in turn allows microfabricated electrokinetic systems to outperform their conventional counterparts by orders of magnitude [3]. The switching components are employed in separation and dispensing systems to inject the sample from the load channel to the separation channel. A typical system employing such switching components is presented in Figure 1, showing a microfluidic system fabricated by etching and bonding in glass.The parameters that determine optimal inje...

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High-Performance DNA Separations in Microchip Electrophoresis Systems

Cited by 15 publications

References 6 publications

Single-step electrohydrodynamic separation of 1–150 kbp in less than 5 min using homogeneous glass/adhesive/glass microchips

Single-step electrohydrodynamic separation of 1–150 kbp in less than 5 min using homogeneous glass/adhesive/glass microchips

Automated Lab-on-a-Chip Analysis of DNA Fragments

Optimization of sample injection components in electrokinetic microfluidic systems

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