WO2003058228A1 - Microplaquette pour electrophorese - Google Patents

Microplaquette pour electrophorese Download PDF

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Publication number
WO2003058228A1
WO2003058228A1 PCT/JP2002/013410 JP0213410W WO03058228A1 WO 2003058228 A1 WO2003058228 A1 WO 2003058228A1 JP 0213410 W JP0213410 W JP 0213410W WO 03058228 A1 WO03058228 A1 WO 03058228A1
Authority
WO
WIPO (PCT)
Prior art keywords
electrophoresis
sample
microchannel
microchip
quantifying
Prior art date
Application number
PCT/JP2002/013410
Other languages
English (en)
Japanese (ja)
Inventor
Minoru Adachi
Isamu Takeuchi
Yoshinobu Baba
Original Assignee
Cluster Technology Co., Ltd.
Millennium Gate Technology Co., Ltd.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Cluster Technology Co., Ltd., Millennium Gate Technology Co., Ltd. filed Critical Cluster Technology Co., Ltd.
Priority to AU2002367338A priority Critical patent/AU2002367338A1/en
Publication of WO2003058228A1 publication Critical patent/WO2003058228A1/fr

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416—Systems
    • G01N27/447—Systems using electrophoresis
    • G01N27/44756—Apparatus specially adapted therefor
    • G01N27/44791—Microapparatus

Definitions

  • the present invention relates to a microchip for electrophoresis.
  • Microelectrophoresis for electrophoresis for measuring or quantifying trace chemicals or biological components.
  • the present invention relates to a tip, an electrophoresis apparatus including the microchip, and a method for detecting or quantifying a trace amount of a chemical substance or a biological component using the electrophoresis apparatus.
  • microchip-based genomic analysis technologies such as DNA chips, microarrays, and microcapillary electrophoresis chips are rapidly advancing, and devices have already been sold.
  • microchip-type capillary array electrophoresis in which 96 channels are arrayed has been developed (Shi, Y. et al., Anal. Chera. , 7 1: 5354-5361, 1999; Baba et al., Experimental Medicine, 18: 12 (extra number) 1595-1601, 2000).
  • the microchannel array is arranged radially, and the center can be scanned while rotating with a laser.
  • This structure requires wiring and a voltage controller to control the potential for each sample, and a small-sized microchip eventually becomes a large-scale device. Also, a large amount of sample to be supplied to the sample amount adjusting structure is required. Therefore, there is a need for a microchip for electrophoresis that is relatively compact and quantitative, and can be analyzed with a small amount of sample. Disclosure of the invention
  • the present invention has been made to solve the above problems, and the present inventors have developed a microchip for electrophoresis having no structure for adjusting a sample amount by using a droplet discharge device having a fine structure. Have been found to be available, and have completed the present invention.
  • the present invention provides a microchip for electrophoresis for analyzing or quantifying a chemical substance or a biological component.
  • the microchip has a width of 200 ⁇ or less and a depth of 100 ⁇ m / zm or less. It has a channel, does not have a microchannel for sample loading using an intersection of microchannels, and has electrodes connectable to a power source at both ends of the microphone opening channel.
  • the microchip further includes a sample loading hole having an opening having a size of 200 111 or less.
  • the microchannel contains a support for electrophoresis.
  • a gel capable of separating the sample.
  • a microstructure capable of separating samples in electrophoresis is provided in the microchannel.
  • the surfaces of the microchannel and the sample loading hole are water-repellent.
  • the present invention also provides an electrophoresis apparatus for analyzing or quantifying a chemical substance or a biological component, comprising any one of the microchips described above, a voltage controller for electrophoresis, and a droplet discharge device having a fine structure. I do.
  • the droplet discharging device is a device having an ink jet head capable of discharging a sample of 10 picoliters (pL) or less.
  • the present invention further provides a method for analyzing or quantifying a chemical substance or a biological component by electrophoresis, the method comprising the steps of: providing a microchannel or a sample loading hole in a microchip of any of the above microchips; Discharging a sample using a droplet discharging device; electrophoresis of the discharged sample in the microchannel; and detecting or quantifying a sample separated by the electrophoresis.
  • FIG. 1 is a conceptual diagram of a top view of a microchip having a sample loading hole according to the present invention, in which (a) is an example in which microchannels are arranged in parallel, (b) is arranged in columns, and (c) is arranged radially. Is shown.
  • FIG. 2 is a cross-sectional view (a) along the line AA and a cross-sectional view (b) along the line BB of the microchip having the sample loading hole of the present invention shown in FIG. 1 (a).
  • the microchip for electrophoresis of the present invention is a microchip for electrophoresis for analyzing or quantifying a chemical substance or a biological component, and the intersection of the microchannels that a conventional microchip for electrophoresis has. A small amount of sample can be electrophoresed in an accurate amount without the use of a sample volume adjustment structure that utilizes the method.
  • the electrophoresis microchip of the present invention will be described.
  • the substrate of the microchip for electrophoresis of the present invention is not particularly limited as long as it is a material usually used for electrophoresis, and examples thereof include glass, fused quartz, crystal quartz, silica, and plastic. It is preferable to use a non-conductive material such as glass and fused silica because a relatively high voltage can be applied.
  • microchannels described in detail below are formed by a fine processing technique. For example, it can be formed by irradiating synchrotron radiation, ion beam, microwave laser, femtosecond laser, and the like. Alternatively, it can be formed by optical lithography and chemical etching.
  • the substrate may be used as it is, or may be used after subjecting the surface to various treatments according to the purpose.
  • the surface treatment method for example, sputtering, CVD (microwave plasma vapor phase synthesis), ion plating, electroless plating, and the like can be mentioned.
  • CVD microwave plasma vapor phase synthesis
  • ion plating electroless plating
  • the surface treatment method for example, sputtering, CVD (microwave plasma vapor phase synthesis), ion plating, electroless plating, and the like can be mentioned.
  • CVD microwave plasma vapor phase synthesis
  • ion plating electroless plating
  • electroless plating electroless plating
  • the microchip for electrophoresis of the present invention is provided with a microchannel for separating and detecting a sample by electrophoresis.
  • the microchannel may be groove-shaped, slit-shaped or tunnel-shaped.
  • the shape of the cross section of the microchannel is not particularly limited, and is appropriately determined depending on the purpose, and may be a concave shape, a U shape, a V shape, or the like.
  • the microchannel can be shaped as shown in FIG. 2 (a).
  • the width of the microchannel is less than 200 / xm and the maximum width is 0.3 ⁇ ! 200200 // m, more preferably 12 ⁇ m to 150 ⁇ m.
  • the depth is ⁇ ⁇ ⁇ or less, preferably 50 ⁇ or less, more preferably 0.3 / zm to 50 ⁇ , and 12 ⁇ ! More preferably, it is 50.
  • the length can be determined as appropriate depending on the sample to be analyzed, the medium used, and the like.
  • the number and arrangement of the microchannels are not particularly limited, but a plurality of microchannels are preferably provided. For example, they can be arranged arbitrarily, such as parallel, tandem, radial, etc. as shown in (a) to (c) of FIG.
  • Droplet force discharged from a droplet discharge device having a fine structure into a microchannel The droplet force adheres to the inner wall of the microchannel due to surface tension, and may not reach the electrophoresis channel in the microchannel.
  • the surface of the microchannel or the surface of the microchip may be subjected to a water-repellent treatment as necessary.
  • the water-repellent treatment may be performed by any method. For example, a method in which a resin film is formed on a substrate and a water-repellent resin layer is formed on the resin film by a physical vapor deposition method is used. No.
  • the microchip of the present invention treated in this way can precisely arrange particularly a very small amount of sample.
  • the microchip for electrophoresis of the present invention includes a microphone such as that found in conventional products. There is no microchannel for sample loading using the intersection of the low channels. Therefore, the microchip of the present invention can be provided as a microarray in which a large number of microchannels are formed on a microchip having a fixed area. Further, microchannels for other uses may be provided according to the purpose. Further, the microchannel may be branched in the middle as necessary.
  • An electrophoresis medium is provided inside the electrophoresis microchannel.
  • the medium include gels, porous polymers, aqueous solutions, and microstructures.
  • Microstructure refers to a three-dimensional structure (Baba et al., Supra) processed by ultra-fine processing technology, which may be engraved in the microchannel in advance. The medium can be appropriately selected depending on the sample to be separated and analyzed.
  • Samples that can be separated and analyzed include, but are not limited to, chemical substances and biological components, and specifically include amino acids, peptides, proteins, nucleic acids (DNA, RNA), polysaccharides, and Alkyroy. And charged substances such as dyes and dyes. Even a neutral substance can be used as a sample by selecting a medium that can be converted to an ionic state.
  • a medium that can be converted to an ionic state.
  • DNA and protein can be suitably analyzed.
  • the medium is preferably a polyacrylamide gel, agarose gel, or the like.
  • the microchip substrate for electrophoresis of the present invention may be provided with a sample loading hole for placing a sample.
  • Sample loading holes are provided on the microchannel to facilitate sample placement in the microchannel.
  • the shape, size and depth of the sample loading hole to be provided is not particularly limited, and may be determined according to the sample amount.
  • the sample loading hole can easily dispense 10 pL into the hole, and is shaped and sized so that the discharged sample reaches the flow path of the microchannel electrophoresis. It is.
  • the cross section BB passing through the sample loading hole is as shown in Fig. 2 (b). In other words, it has a slightly larger opening than the microchannel shown in Fig. 2 (a), and has a shape that allows easy placement of the sample.
  • the sample loading hole is preferably subjected to a water-repellent treatment in the same manner as the above-described microchannel, in that the sample can surely reach the flow channel of the microchannel.
  • the shape of the opening may be, for example, a circle, a square, a diamond, or an ellipse.
  • a through hole having the same shape and size as the opening of the sample loading hole is formed in the cover plate.
  • the microchannel is typically provided at both ends with electrodes connectable to a power supply.
  • the sample By using the side on which the sample is loaded as the cathode and the other end as the anode, the sample can be moved as in the usual electrophoresis.
  • the material of the electrode is not particularly limited as long as it is usually used as an electrode.
  • the electrodes may be individually connected to the power supply, but the wires on the cathode side and the anode side may be combined into one, and each microchannel may be connected in parallel to the power supply. In this way, the connection to the power supply is easy, and the wiring of the microchip itself can be simplified and the microchip can be made compact. (Droplet ejection device with fine structure)
  • the droplet discharge device having a fine structure may be any device that can discharge a sample of 10 pL or less, preferably 4 pL or less with high accuracy.
  • the droplet discharge device having an ink jet head as described in Semiconductor FPD World 2001. 6, 154-156.
  • it may be a bubble jet (registered trademark) device, a micropump, a micropitter, or the like.
  • the sample separated by electrophoresis can be detected by a commonly used detection method. Typically, UV absorption, fluorescence, chemiluminescence, change in refractive index, etc. are measured. In the present invention, fluorescence detection is preferred because a small amount of sample is measured. In addition, since the sample can be arranged in an accurate amount, quantification by measuring the intensity of the detected signal is possible. Industrial availability '14
  • the microchip for electrophoresis of this invention can arrange
  • there is no structure for adjusting the sample amount there is no wasteful sample placement on the chip compared to the conventional one, and measurement can be performed with a small sample amount, and cost performance is good.
  • the simple structure of the microchip allows many microchannels to be placed on the microchip. Thus, a compact and highly accurate microchip for electrophoresis can be easily provided.

Abstract

L'invention concerne une microplaquette pour électrophorèse, qui permet d'analyser ou de déterminer des substances chimiques ou des composants biologiques. Cette microplaquette, qui comporte des microcanaux ayant chacun jusqu'à 200 νm de largeur et 100 νm de profondeur, présente un microcanal destiné au chargement d'échantillon qui n'utilise pas l'intersection des microcanaux susmentionnés et dont les extrémités opposées comportent des électrodes susceptibles d'être reliées respectivement à une source d'alimentation. Il en résulte une microplaquette pour électrophorèse comparativement compacte, à fonction d'évaluation quantitative, offrant la possibilité d'une analyse sur la base de faibles quantités d'échantillon.
PCT/JP2002/013410 2001-12-28 2002-12-20 Microplaquette pour electrophorese WO2003058228A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2002367338A AU2002367338A1 (en) 2001-12-28 2002-12-20 Electrophoresis-use microchip

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2001401033A JP2005164242A (ja) 2001-12-28 2001-12-28 電気泳動用マイクロチップ
JP2001-401033 2001-12-28

Publications (1)

Publication Number Publication Date
WO2003058228A1 true WO2003058228A1 (fr) 2003-07-17

Family

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Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2002/013410 WO2003058228A1 (fr) 2001-12-28 2002-12-20 Microplaquette pour electrophorese

Country Status (3)

Country Link
JP (1) JP2005164242A (fr)
AU (1) AU2002367338A1 (fr)
WO (1) WO2003058228A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012017515A1 (fr) * 2010-08-03 2012-02-09 ミライアル株式会社 Dispositif à micro-canal

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP6017793B2 (ja) 2012-02-09 2016-11-02 ローム株式会社 マイクロチップ

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH08327593A (ja) * 1995-05-29 1996-12-13 Shimadzu Corp キャピラリー電気泳動装置
JPH11508360A (ja) * 1995-06-21 1999-07-21 フアーマシア・バイオテツク・アー・ベー フロースルーサンプリングセルおよびその使い方
JP2000246805A (ja) * 1999-03-04 2000-09-12 Kawamura Inst Of Chem Res マイクロケミカルデバイスの製造法
JP2000310614A (ja) * 1999-02-26 2000-11-07 Hitachi Chem Co Ltd 電気泳動用チップとその製造方法、該電気泳動用チップを用いた電気泳動装置及び荷電性物質の分離方法
JP2001056286A (ja) * 1999-08-18 2001-02-27 Shimadzu Corp 測定用セル
JP2001507441A (ja) * 1995-06-08 2001-06-05 ビジブル ジェネティクス インコーポレイテッド 核酸および他の荷電された分子を移動させそして分離するための微小電気泳動チップ
JP2001228159A (ja) * 2000-02-14 2001-08-24 Seiko Epson Corp マイクロリアクタ
JP2001515216A (ja) * 1997-08-13 2001-09-18 シーフィード 流体試料を操作するための微小構造体
JP2002207031A (ja) * 2001-01-11 2002-07-26 Shimadzu Corp マイクロチャンネル型チップ

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH08327593A (ja) * 1995-05-29 1996-12-13 Shimadzu Corp キャピラリー電気泳動装置
JP2001507441A (ja) * 1995-06-08 2001-06-05 ビジブル ジェネティクス インコーポレイテッド 核酸および他の荷電された分子を移動させそして分離するための微小電気泳動チップ
JPH11508360A (ja) * 1995-06-21 1999-07-21 フアーマシア・バイオテツク・アー・ベー フロースルーサンプリングセルおよびその使い方
JP2001515216A (ja) * 1997-08-13 2001-09-18 シーフィード 流体試料を操作するための微小構造体
JP2000310614A (ja) * 1999-02-26 2000-11-07 Hitachi Chem Co Ltd 電気泳動用チップとその製造方法、該電気泳動用チップを用いた電気泳動装置及び荷電性物質の分離方法
JP2000246805A (ja) * 1999-03-04 2000-09-12 Kawamura Inst Of Chem Res マイクロケミカルデバイスの製造法
JP2001056286A (ja) * 1999-08-18 2001-02-27 Shimadzu Corp 測定用セル
JP2001228159A (ja) * 2000-02-14 2001-08-24 Seiko Epson Corp マイクロリアクタ
JP2002207031A (ja) * 2001-01-11 2002-07-26 Shimadzu Corp マイクロチャンネル型チップ

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
YOSHINOBU BABA: "Micro chip nano chip technology ni yoru jisedai DNA kaiseki gijutsu no kaihatsu", EXPERIMENTAL MEDICINE, vol. 18, no. 12, 2000, pages 41 - 47, XP002975361 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012017515A1 (fr) * 2010-08-03 2012-02-09 ミライアル株式会社 Dispositif à micro-canal

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AU2002367338A1 (en) 2003-07-24
JP2005164242A (ja) 2005-06-23

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