Electrophoresis of DNA in micro-pillars fabricated in polydimethylsiloxane

Inatomi, Ken-ichi; Izuo, Shinichi; Lee, Sang-seok; Ohji, Hiroshi; Shiono, Satoru

doi:10.1016/s0167-9317(03)00361-7

Cited by 35 publications

(41 citation statements)

References 14 publications

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“…However, each time, exclusively obstacles in the dimension of the radius of gyration of the investigated DNA molecules were used, the same mobility than in our case has been reported (Inatomi et al, 2003;Kaji et al, 2003). This effect, however, not completely understood, is currently under investigation experimentally as well as by theoretical simulations.…”

Section: Dna Injection and Separationsupporting

confidence: 72%

“…One-dimensional (Slentz et al, 2001) and three-dimensional (Slentz et al, 2003) electrochromatography of a fluorescein isothiocyanate (FITC)-labeled tryptic digest of BSA have also been demonstrated in PDMS microchips. Applications to DNA include restriction fragment analysis (Effenhauser et al, 1997), as well as free solution DNA migration studies (Duong et al, 2003;Inatomi et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

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Towards single molecule analysis in PDMS microdevices: from the detection of ultra low dye concentrations to single DNA molecule studies

Ros

Hellmich

Duong

et al. 2004

Journal of Biotechnology

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In this paper, we report on the performance of electrophoretical separation and laser-induced fluorescence (LIF) detection of dyes and fluorescently labeled biomolecules in poly(dimethylsiloxane) (PDMS) microdevices. The dyes fluorescein and fluorescein isothiocyanate (FITC) have been separated effectively in nM concentrations. Fluorescein injections gave linear concentration response in the range from 4 to 100 pM. As ultimate detection sensitivity, 100 fM injected fluorescein was obtained. Further, 100 fM injected fluorescein could be detected. This is to our knowledge the smallest electrokinetically injected dye concentration detected on a microchip. Injection studies of fluorescently labeled avidin revealed a theoretical detection limit of 25 nM for laser-induced fluorescence detection in good agreement with separations in glass chips. Furthermore, the injection of several and even one single DNA molecule using a PDMS cross injector has been demonstrated as well as free solution separation of -and T2-DNA (60 pM each) in periodically structured channels.

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Section: Dna Injection and Separationsupporting

confidence: 72%

Section: Introductionmentioning

confidence: 99%

Towards single molecule analysis in PDMS microdevices: from the detection of ultra low dye concentrations to single DNA molecule studies

Ros

Hellmich

Duong

et al. 2004

Journal of Biotechnology

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“…Once the lateral deflections of the microposts are obtained by image acquisition and analysis, CTFs can be determined based on the beam theory [Timoshenko and Woinowskey-Kreiger, 1959]. Besides CTF measurement, micropost arrays have also been used for assembling freely suspended two-dimensional actin cortex network [Roos et al, 2003] and myofibrils [Zhao and Zhang, 2006], electrophoresis separation of DNA [Inatomi et al, 2003], and studying dynamics of molecular motor-driven microtubule gliding and microtubule network formation .…”

Section: Introductionmentioning

confidence: 99%

Development of micropost force sensor array with culture experiments for determination of cell traction forces

Xie

Starr

et al. 2007

Cell Motil. Cytoskeleton

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Cell traction forces (CTFs) are critical for cell motility and cell shape maintenance. As such, they play a fundamental role in many biological processes such as angiogenesis, embryogenesis, inflammation, and wound healing. To determine CTFs at the sub-cellular level with high sensitivity, we have developed high density micropost force sensor array (MFSA), which consists of an array of vertically standing poly(dimethylsiloxane) (PDMS) microposts, 2 microm in diameter and 6 microm in height, with a center-to-center distance of 4 microm. In combination with new image analysis algorithms, the MFSA can achieve a spatial resolution of 40 nm and a force sensitivity of 0.5 nN. Culture experiments with various types of cells showed that this MFSA technology can effectively determine CTFs of cells with different sizes and traction force magnitudes.

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“…That is, this separation is accomplished by molecular sieving effect microfluidics. Analysis of the motion of a single DNA molecules have been previously carried out on device made of polydimethylsiloxane(PDMS) 6 that has a micro-channel in which arranged in a hexagonal lattice. Electrophoresis of DNA in the pillars showed the average speed of DNA migration was dependent on its lengths.…”

Section: Different Methods Used For Dna Separationmentioning

confidence: 99%

Microminiaturized electrophoresis DNA separator using MEMS

2005

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Conventional DNA separation procedures involve large sizes, high voltages and are unfit for large molecules. Proposed set-up constitutes two fold separation techniques, porous filter for fractionating strands on size and final run through microchannels (agarose, buffer solution) viscous enough for DNA electrophoresis. For arriving at the final set-up all the physical contradictions like voltage, viscosity of the fluid, length of the channel were analyzed.The physical set up consists around 20 microchannels (varying diameters) positioned at 500um (centre-to-centre spacing) assembling entire device within 1cm. Polymerase Chain Reaction (PCR) treated DNA assays, are fed to microchannel entrances. Mixture of DNA strands are then passed through magnetic filters. Filtering property of the filters is adjusted by regulating corresponding magnetic field strengths. Smallest strands pass through small pored filter, owing to high velocity (in electrophoresis), thus categorization being done. Proposal replaces conventional apparatus by miniaturized equipment, in ideal case disposable. Miniaturization reduced voltages requirement, solving high-voltage handling problems. Proposed apparatus can fractionate large (>200kbp) molecules and even organic molecules.

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Electrophoresis of DNA in micro-pillars fabricated in polydimethylsiloxane

Cited by 35 publications

References 14 publications

Towards single molecule analysis in PDMS microdevices: from the detection of ultra low dye concentrations to single DNA molecule studies

Towards single molecule analysis in PDMS microdevices: from the detection of ultra low dye concentrations to single DNA molecule studies

Development of micropost force sensor array with culture experiments for determination of cell traction forces

Microminiaturized electrophoresis DNA separator using MEMS

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