A nanoliter-scale nucleic acid processor with parallel architecture

Hong, Jong Wook; Studer, Vincent; Hang, Giao; Anderson, William F.; Quake, Stephen R.

doi:10.1038/nbt951

Cited by 437 publications

(359 citation statements)

References 18 publications

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“…55 After the cells had been lysed, the lysate was flushed over an affinity column packed with oligo-dT polymer magnetic beads (Figure 4). Reverse transcriptase PCR was performed directly on the beads after the column was washed; as few as two cells could be detected.…”

Section: Integrated Sample-preparation Systemsmentioning

confidence: 99%

Purification of Nucleic Acids in Microfluidic Devices

et al. 2008

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Section: Integrated Sample-preparation Systemsmentioning

confidence: 99%

Purification of Nucleic Acids in Microfluidic Devices

et al. 2008

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“…The circular channels in the center are the rotary mixers and the thicker black marks are mechanical valves. 73 beneficial experimental basis for systems biology because: (a) Micro-scale fluid flow enables precise and high-resolution microenvironment control; (b) Microfluidic devices are generally compatible with imaging and microscopy techniques; (c) The ability to upscale and array microfluidic devices facilitates high-throughput experimentation, a necessary ability when faced with the extraordinary complexity of biological systems.…”

Section: Discussionmentioning

confidence: 99%

Microfluidics-based systems biology

2006

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Systems biology seeks to develop a complete understanding of cellular mechanisms by studying the functions of intra-and inter-cellular molecular interactions that trigger and coordinate cellular events. However, the complexity of biological systems causes accurate and precise systems biology experimentation to be a difficult task. Most biological experimentation focuses on highly detailed investigation of a single signaling mechanism, which lacks the throughput necessary to reconstruct the entirety of the biological system, while high-throughput testing often lacks the fidelity and detail necessary to fully comprehend the mechanisms of signal propagation. Systems biology experimentation, however, can benefit greatly from the progress in the development of microfluidic devices. Microfluidics provides the opportunity to study cells effectively on both a single-and multi-cellular level with high-resolution and localized application of experimental conditions with biomimetic physiological conditions. Additionally, the ability to massively array devices on a chip opens the door for high-throughput, high fidelity experimentation to aid in accurate and precise unraveling of the intertwined signaling systems that compose the inner workings of the cell.

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“…It is also possible to modify the array design such that each individual chamber has a separate outlet, allowing the collection of fluidic samples for biochemical analysis (e.g., PCR, ELISA, HPLC, mass spectroscopy). Automation of array operation and data collection can be achieved by integrating previously developed microfluidic technology, allowing massively multiplexed cell level analysis of individual cells (Hong et al, 2004;Thorsen et al, 2002). By enabling biologists with a robust method to control the fluidic environments of an array of cell culture units, the precision and accuracy of quantitative cell experiments will be greatly improved.…”

Section: Discussionmentioning

confidence: 99%

Nanoliter scale microbioreactor array for quantitative cell biology

Lee

Hung

Rao

et al. 2005

Biotech & Bioengineering

203

183

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A nanoliter scale microbioreactor array was designed for multiplexed quantitative cell biology. An addressable 8 Â 8 array of three nanoliter chambers was demonstrated for observing the serum response of HeLa human cancer cells in 64 parallel cultures. The individual culture unit was designed with a ''C'' shaped ring that effectively decoupled the central cell growth regions from the outer fluid transport channels. The chamber layout mimics physiological tissue conditions by implementing an outer channel for convective ''blood'' flow that feeds cells through diffusion into the low shear ''interstitial'' space. The 2 mm opening at the base of the ''C'' ring established a differential fluidic resistance up to 3 orders of magnitude greater than the fluid transport channel within a single mold microfluidic device. Three-dimensional (3D) finite element simulation were used to predict fluid transport properties based on chamber dimensions and verified experimentally. The microbioreactor array provided a continuous flow culture environment with a Peclet number (0.02) and shear stress (0.01 Pa) that approximated in vivo tissue conditions without limiting mass transport (10 s nutrient turnover). This microfluidic design overcomes the major problems encountered in multiplexing nanoliter culture environments by enabling uniform cell loading, eliminating shear, and pressure stresses on cultured cells, providing stable control of fluidic addressing, and permitting continuous on-chip optical monitoring. ß 2005 Wiley Periodicals, Inc.

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A nanoliter-scale nucleic acid processor with parallel architecture

Cited by 437 publications

References 18 publications

Purification of Nucleic Acids in Microfluidic Devices

Purification of Nucleic Acids in Microfluidic Devices

Microfluidics-based systems biology

Nanoliter scale microbioreactor array for quantitative cell biology

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