[논문]Biochemical Reactions on a Microfluidic Chip Based on a Precise Fluidic Handling Method at the Nanoliter Scale

Lee, Chang-Soo; Lee, Sang-Ho; Kim, Yun-Gon; Choi, Chang-Hyoung; Kim, Yong-Kweon; Kim, Byung-Gee

doi:10.1007/bf02931899

[국내논문] Biochemical Reactions on a Microfluidic Chip Based on a Precise Fluidic Handling Method at the Nanoliter Scale 원문보기

Biotechnology and bioprocess engineering : Bbe, v.11 no.2, 2006년, pp.146 - 153

Lee, Chang-Soo (Department of Chemical Engineering, Chungnam National University) , Lee, Sang-Ho (School of Electrical Engineering and Computer Science, Seoul National University) , Kim, Yun-Gon (School of Chemical and Biological Engineering and Institute of Molecular Biology and Genetics, Seoul National University) , Choi, Chang-Hyoung (Department of Chemical Engineering, Chungnam National University) , Kim, Yong-Kweon (School of Electrical Engineering and Computer Science, Seoul National University) , Kim, Byung-Gee (School of Chemical and Biological Engineering and Institute of Molecular Biology and Genetics, Seoul National University)

Abstract ▼ AI-Helper

A passive microfluidic delivery system using hydrophobic valving and pneumatic control was devised for microfluidic handling on a chip. The microfluidic metering, cutting, transport, and merging of two liquids on the chip were correctly performed. The error range of the accuracy of microfluid metering was below 4% on a 20 nL scale, which showed that microfluid was easily manipulated with the desired volume on a chip. For a study of the feasibility of biochemical reactions on the chip, a single enzymatic reaction, such as ${\beta}-galactosidase$ reaction, was performed. The detection limit of the substrate, i.e. fluorescein $di-{\beta}-galactopyranoside$ (FDG) of the ${\beta}-galactosidase$ (6.7 fM), was about 76 pM. Additionally, multiple biochemical reactions such as in vitro protein synthesis of enhanced green fluorescence protein (EGFP) were successfully demonstrated at the nanoliter scale, which suggests that our microfluidic chip can be applied not only to miniaturization of various biochemical reactions, but also to development of the microfluidic biochemical reaction system requiring a precise nano-scale control.

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가설 설정

The fabrication process of the microfluidic chip. A. Photoresist (PR) patterning for the definition of a hydrophobic region. B.

제안 방법

A Kriiss GIO contact angle analyzer (Kriiss GmbH, Hamburg, Germany) was used to investigate the potential surface wettability and surface tension of hydrophobic thin film. All of the measurements were carried out at room temperature and ambient humidity.
3. The principle of microfluidic handling; in this experiment, color colloidal inks were used to obtain clear images. A.
To biveslig사te the accuracy of the proposed microfluidic handling method on a chip, the microscopic images of the final metered droplet were captured using a CCD camera. The captured images were analyzed using the image analysis software, Kontron KS Lite, in order to determine the droplet area. The measured area was converted to a corresponding volume by multiplying the value of the microchannel depth, 50 μm.
In order to estimate the feasibility of the proposed microfluidic handling method for a biochemical reaction, we examined the P-galactosidase reaction performed on a microfluidic chip. In this experiment, because p-galactosidase hydrolyzes the non-fluorescent FDG to fluorescein via the intermediate compound, fluorescein mono-P-galactopyranoside (FMG), the P-galactosidase reaction was easily detected by fluorescent microscopy.
development of “lab on a chip”. In this study, we used the microfluidic handling method to manipulate the motion of microfluid, performing injection, metering, cutting, transport, and merging on a chip. Based on the surface patterning for different wettability and pneumatic pressure balance in chip design, the microfluid is discretely metered to the desired volume depending on the length of the metering channel.

이론/모형

Figs. 1A to C show the hydrophobic thin film patterning ono glass substrate using the lift-off method. First, a 2,000 A aluminum film was patterned using a phosphoric acid-based etchant and a photoresist (PR) mask through the thermal evaporation process (Fig.

성능/효과

The measured area was converted to a corresponding volume by multiplying the value of the microchannel depth, 50 μm. In thirty metering runs using a criterion of 20 nL, the accuracy of the metering showed about 4% error (mean = 20.168 nL, standard deviation = 0.730 nL; Fig. 4), which proved that our proposed microfluidic handling method has reliable accuracy at the nanoliter scale.
7). The result showed that the limit of detection could be lowered to five orders of magnitude over a conventional assay, and the amount of consumed reagents could be reduced from microscale to nanoscale. Furthermore, if we adopted a mixing part or mixing method for performing a rapid homogeneous reaction on a chip, the reaction would be faster and the limit of detection would be decreased to a higher degree on a microfluidic chip.

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