Large scale patterning of hydrogel microarrays using capillary pinning

Gumuscu, Burcu; Bomer, Johan G.; Berg, Albert van den; Eijkel, Jan C.T.

doi:10.1039/c4lc01350f

Cited by 26 publications

(26 citation statements)

References 35 publications

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“…The reservoir volume is 2000 times larger than the inner volume of the microchip and the buffer solution was frequently refreshed. The μGEL device has a calculated throughput of 0.18 ng of molecules per hour at the DNA input concentration of 12.5 ng μL − 1 and is comparable to previously reported microfabricated devices 20,31,32 .…”

Section: Resultssupporting

confidence: 50%

“…The observed reorientation times were found to be in good agreement with theory, calculated as 0.2 and 0.4 s, respectively. At frequencies far below f or;2 ¼ 1 2tor;2 μE2 2LDNA , the DNA fragments will quickly reorient along the new electric field direction and will spend most of the application time moving in steady state along the field 20 . Different from the predictions of the switchback mechanism, we still observe separation at these frequencies.…”

Section: Resultsmentioning

confidence: 99%

“…The microchip consists of a 10 mm by 10 mm chamber connected to buffer reservoirs via parallel microchannel arrays (50 μm × 10 mm, 50 μm periodicity) on four sides. The microchannels provide a high-resistance area, preventing current leakage from the separation chamber and generating a fairly uniform electric field (Supplementary Figure S1) 20,22 .…”

Section: Resultsmentioning

confidence: 99%

“…The mixture was boiled in a microwave and pipetted into the microchip that was warmed to 70°C on a hot plate. The empty space in the microchip was filled entirely by capillary forces 20,21 . The microchip was then immersed in a buffer solution overnight.…”

Section: Microchip Fabricationmentioning

confidence: 99%

“…Finally, both top and bottom layers were cleaned and thermally bonded at 1080°C. The bonded wafers were then diced into individual microchips 20 .…”

Section: Microchip Fabricationmentioning

confidence: 99%

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Exploiting biased reptation for continuous flow preparative DNA fractionation in a versatile microfluidic platform

et al. 2017

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A new approach is presented for preparative, continuous flow fractionation of sub-10-kbp DNA fragments, which exploits the variation in the field-dependent mobility of the DNA molecules based on their length. Orthogonally pulsed electric fields of significantly different magnitudes are applied to a microchip filled with a sieving matrix of 1.2% agarose gel. Using this method, we demonstrate a high-resolution separation of 0.5, 1, 2, 5, and 10 kbp DNA fragments within 2 min. During the separation, DNA fragments are also purified from other ionic species. Preparative fractionation of sub-10-kbp DNA molecules plays an important role in second-generation sequencing. The presented device performs rapid high-resolution fractionation and it can be reliably manufactured with simple microfabrication procedures. This method has the advantages of simplicity, versatility, and reproducibility. However, it suffers from long processing times on the order of a few tens of hours 2,3 . For instance, the Bio-Rad CHEF-DR device performs fractionation of 5-120 kbp DNA using pulsed-field gel electrophoresis (PFGE) in 25 h (Refs. 4,5). Trends toward second-generation sequencing motivate the replacement of standard gel electrophoresis with microchip-based systems, which could provide efficient platforms to minimize the processing time and to perform optimal DNA fractionation 6-8 .Micro-and nanofabricated post arrays-that resemble a gel matrix with well-ordered and identical pores-have been integrated in microchip-based systems, increasing both the understanding of DNA separation principles and the fractionation's efficiency and speed. Two decades ago, Volkmuth and Austin 9 introduced a patterned micro-post array in a microfluidic electrophoresis platform, in which DNA fragments were separated by biased reptation under a DC electric field. Later, Duke et al. separated large fragments in a range of 60-135 kbp in a microfabricated array using DNA reorientation-or the 'switchback' principle. Although the switchback principle is similar to what is used in PFGE, the device yields much faster separations owing to its sparse and regular sieving array 10 . In a later work, Kaji et al. studied a three-dimensional (3D) nanopost array as an optimal separation matrix for DNA fragments over a few kbps under a DC electric field, resulting in similarly fast separations due to the sparse matrix 11 .With the aim of increasing the sample throughput and further facilitating the sample recovery, a continuous flow separation was developed. Huang et al. 12 performed continuous flow PFGE in a micromachined post array by applying pulsed electric fields of slightly unequal strengths. DNA fragments ranging between 61 and 209 kbp were separated within 15 s in the 'DNA prism'.

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Section: Resultssupporting

confidence: 50%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Microchip Fabricationmentioning

confidence: 99%

“…Finally, both top and bottom layers were cleaned and thermally bonded at 1080°C. The bonded wafers were then diced into individual microchips 20 .…”

Section: Microchip Fabricationmentioning

confidence: 99%

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Exploiting biased reptation for continuous flow preparative DNA fractionation in a versatile microfluidic platform

et al. 2017

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Desalination by Electrodialysis Using a Stack of Patterned Ion‐Selective Hydrogels on a Microfluidic Device

Gumuscu

Haase

Benneker

et al. 2016

Adv Funct Materials

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This study reports a novel approach for separation of charged species using anion‐exchange hydrogel (AEH) and cation‐exchange hydrogel (CEH) in a microfluidic device. The capillary line pinning technique, which is applied in this study, enables in situ fabrication of alternating AEH and CEH that are placed in confined compartments. Adjacent enriched and depleted streams are obtained in continuous flow when a potential difference is applied over the hydrogel stack. The desalination performance of the microchip is demonstrated at different salt concentrations (0.01 × 10−3–1× 10−3m sodium chloride), potentials (10–100 V), current densities (12–28 A m−2), and liquid flow rates (0–5 µL min−1). It is shown that the microchip is able to remove ≈75% of the salt initially present in the depleted outlet streams at inlet stream concentrations of 1 × 10−3m sodium chloride. Besides desalination, the microchip allows study of ion transport in the ion‐selective hydrogels to elucidate the interplay of transport phenomena at the electrolyte–hydrogel interface during the desalination process.

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Microscale Interfacial Polymerization on a Chip

et al. 2021

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Forming hydrogels with precise geometries is challenging and mostly done using photopolymerization, which involves toxic chemicals, rinsing steps, solvents, and bulky optical equipment. Here, we introduce a new method for in situ formation of hydrogels with a well-defined geometry in a sealed microfluidic chip by interfacial polymerization. The geometry of the hydrogel is programmed by microfluidic design using capillary pinning structures and bringing into contact solutions containing hydrogel precursors from vicinal channels. The characteristics of the hydrogel (mesh size, molecular weight cut-off) can be readily adjusted. This method is compatible with capillary-driven microfluidics, fast, uses small volumes of reagents and samples, and does not require specific laboratory equipment. Our approach creates opportunities for filtration, hydrogel functionalization, and hydrogel-based assays, as exemplified by a rapid, compact competitive immunoassay that does not require a rinsing step.Hydrogels are excellent materials for microfluidic applications in biomedicine and biosensing owing to their compatibility with proteins and numerous solvents, permeability to small chemicals, transparency, potential for functionalization, and programmable stiffness. [1][2][3][4][5] For example, hydrogels have been employed for fabricating membranes in chips for microdialysis, [6,7] filtration, [8] preconcentration of proteins in electrophoresis, [9] or studying solvophoresis [10] and diffusophoresis. [11] Likewise, hydrogels are frequently applied for the immobilization of biomolecular receptors in microfluidics to enable sensitive detection of proteins, [12,13] nucleic acids, [14] or small-molecule targets. [15] Moreover, due to their tunable response to temperature, pH, light, electric and magnetic fields, hydrogels are increasingly being used as microfluidic actuators, which leads to numerous novel applications. [16][17][18]

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Large scale patterning of hydrogel microarrays using capillary pinning

Cited by 26 publications

References 35 publications

Exploiting biased reptation for continuous flow preparative DNA fractionation in a versatile microfluidic platform

Exploiting biased reptation for continuous flow preparative DNA fractionation in a versatile microfluidic platform

Desalination by Electrodialysis Using a Stack of Patterned Ion‐Selective Hydrogels on a Microfluidic Device

Microscale Interfacial Polymerization on a Chip

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