PDMS–glass hybrid microreactor array with embedded temperature control device. Application to cell-free protein synthesis

Yamamoto, Takatoki; Fujii, Teruo; Nojima, Takahiko

doi:10.1039/b205010b

Cited by 113 publications

(76 citation statements)

References 16 publications

(13 reference statements)

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“…14 We have reported in vitro translation in microreactors, 15,16 and have already succeeded in the cell-free translation of GFP in a microfabricated device equipped with a fluorescence microscope, and the in situ detection of the expressed GFP. 17 Further studies are underway to attain this goal.…”

Section: Resultsmentioning

confidence: 99%

Determination of the Termination Efficiency of the Transcription Terminator Using Different Fluorescent Profiles in Green Fluorescent Protein Mutants

et al. 2005

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Section: Resultsmentioning

confidence: 99%

Determination of the Termination Efficiency of the Transcription Terminator Using Different Fluorescent Profiles in Green Fluorescent Protein Mutants

et al. 2005

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“…[16][17][18][19] In such a microfluidic system, pressure or electroosmotic flow can be used to control the transport of the cells together with their surrounding media. [20][21][22] These shrinking laboratories have allowed smaller and smaller samples to be used. A potential limitation, however, is the difficulty in changing the media around the cells under investigation without also losing control of the cells.…”

Section: Introductionmentioning

confidence: 99%

Optical tweezers applied to a microfluidic system

et al. 2004

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We will demonstrate how optical tweezers can be combined with a microfluidic system to create a versatile microlaboratory. Cells are moved between reservoirs filled with different media by means of optical tweezers. We show that the cells, on a timescale of a few seconds, can be moved from one reservoir to another without the media being dragged along with them. The system is demonstrated with an experiment where we expose E. coli bacteria to different fluorescent markers. We will also discuss how the system can be used as an advanced cell sorter. It can favorably be used to sort out a small fraction of cells from a large population, in particular when advanced microscopic techniques are required to distinguish various cells. Patterns of channels and reservoirs were generated in a computer and transferred to a mask using either a sophisticated electron beam technique or a standard laser printer. Lithographic methods were applied to create microchannels in rubber silicon (PDMS). Media were transported in the channels using electroosmotic flow. The optical system consisted of a combined confocal and epi-fluorescence microscope, dual optical tweezers and a laser scalpel.

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“…1. Because the chip design detail and temperature control capability are described in a previous paper, 19 we only briefly explain the microreactor array chip. The microreactor array is a hybrid of two kinds of chips: one is a reaction chamber chip; the other is a temperature control chip as presented schematically in Fig.…”

Section: Microreactor Array Chipmentioning

confidence: 99%

“…We have recently developed a microreactor array chip with an embedded temperature control unit and have demonstrated its performance experimentally by synthesizing fluorescent proteins. 19 For the present study, we evaluated our microreactor array chip by expressing various types of proteins in comparison with a conventional commercially available reaction system to evaluate the feasibility of the microreactor array chip for future high-throughput protein research.…”

Section: 2mentioning

confidence: 99%

Evaluation of Cell-free Protein Synthesis Using PDMS-based Microreactor Arrays

et al. 2008

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As represented by the recent completion of human genome sequencing, the genome sequences of several species have been, or are about to be, revealed. Although all of their sequential information will eventually become available, it will provide little information related to the function of encoded proteins because we can not predict post-translational events merely from the sequence. The next challenge is therefore to elucidate the functions and physiological roles of every encoded protein that is functioning in living cells under various environmental conditions: so-called "proteomes". 1,2Several methods to analyze proteins have been invented during the history of biotechnology; particularly, biochemical analysis is a very reliable method to determine protein functions. Biochemical assay necessarily imposes limitations in realizing highthroughput biochemical analysis in production of vastly numerous proteins because the current technology of protein production is based on in vivo methods that require time and labor because of their use of bacteria. As an alternative, cellfree protein synthesis is a useful and profitable technique to overcome problems associated with in vivo methods. By introducing cell-free formats, we can control many aspects of cellular functions, which simplifies protein production, especially those of cytotoxic, unstable, and insoluble proteins. [3][4][5][6] Commercially available cell-free systems with embedded thermo-control units have been developed, e.g., the Rapid Translation System (Roche Molecular Biochemicals). However, this system is oriented to produce as much as possible efficiently; it is insufficiently optimized for high-throughput protein analysis, because it is impractical to conduct parallel operation with this system from the viewpoint of space efficiency, automatic operation, and reagent consumption in order to improve the throughput of protein production.Because researches on micro total analysis system (μ-TAS) and lab-on-a-chip have advanced, several applications have emerged at the downstream for production of small amounts of proteins. For example, chip-based capillary electrophoresis in combination with micro injector for mass spectrometer 7-11 and enormous sample preparation devices for high-throughput mass spectrometry 12 have enabled researchers to perform protein analysis with much smaller amounts than those used in conventional methods.Micro-crystallizer for X-ray crystallography and NMR analysis have also been developed for high-throughput crystallization from only small amounts of protein. 13,14 For that reason, large amounts of protein are no longer required anymore for analysis; nevertheless, a persistent problem is how to prepare many kinds of proteins. Miniaturization of a cell-free protein synthesis system has great potential to provide useful platforms to produce many kinds of proteins. Physico-chemical phenomena in micro-sized channels and reaction chambers intrinsically involve a short diffusion length, which engenders rapid thermal conduction and mixing...

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PDMS–glass hybrid microreactor array with embedded temperature control device. Application to cell-free protein synthesis

Cited by 113 publications

References 16 publications

Determination of the Termination Efficiency of the Transcription Terminator Using Different Fluorescent Profiles in Green Fluorescent Protein Mutants

Determination of the Termination Efficiency of the Transcription Terminator Using Different Fluorescent Profiles in Green Fluorescent Protein Mutants

Optical tweezers applied to a microfluidic system

Evaluation of Cell-free Protein Synthesis Using PDMS-based Microreactor Arrays

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