JP2003240755A - Microchip comprising transparent hydrocarbon-based polymer - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a microchip made of a resin with a microchannel with a channel sealed by an upper substrate which is transparent and absorbs little in ultraviolet region, is minute and has satisfactory surface smoothness, and is high in dimension accuracy. <P>SOLUTION: The microchip for optically measuring a liquid sample has at least both a lower substrate with the microchannel through which the liquid sample flows and the upper substrate made of a flat film. At least the lower substrate is made of a transparent hydrocarbon-based polymer expressed by Fig. (1). At least the surface of the lower substrate on the side with the channel is treated with oxygen plasma, and then the lower substrate and the upper substrate are subjected to thermocompression bonding at temperatures lower than the glass-transition temperature (Tg) of the transparent hydrocarbon-based polymer to obtain the microchip. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a resin-made microchip used for simple analysis and detection of a small amount of sample such as DNA and protein.

[0002]

2. Description of the Related Art In recent years, attention has been paid to a technique for performing a chemical reaction, separation and analysis in a microscopic area. Specific application examples of such technology include bedside diagnosis that performs measurements necessary for medical diagnosis near the patient, analysis of harmful substances in waste in rivers and soils near the measurement target, and further food cooking and harvesting. , Point Of
Care (hereinafter referred to as POC) analysis is considered, and the development of detection methods and devices for it is an important issue. In such POC analysis, a trace amount analysis method that is inexpensive, simple, and can be performed in a short time is required.

Conventionally, GCMS and LCMS, in which components are separated by capillary gas chromatography (CGC), capillary liquid chromatography (CLC), etc. and then detected by a mass spectrometer, have been widely used for the analysis and detection of a small amount of sample. It was However, GCMS and LCMS
The mass spectrometer of the detector is large, and P
Difficult to use OC. Moreover, since contact with a sample derived from a patient such as blood becomes a problem, the equipment in such a case is a target of infectious waste and basically required to be disposable.

In order to solve the above problems, 10 cm
To a few cm square or less, such as a glass chip (hereinafter referred to as a microchip), a groove is formed on the surface thereof, and a minute sample is analyzed by using the separation and reaction in the groove.
Research on AS (miniaturized total analysis system) is under way.

With μ-TAS, the amount of sample, the amount of reagent necessary for detection, the amount of waste such as consumables used for detection, and the amount of waste liquid are all small, and the time required for detection is generally short. There is an advantage. The microchip used in the above microanalyzer has the following points from the viewpoint of its workability and accuracy:
Inorganic materials such as a glass plate, a quartz plate, and a silicon plate are mainly used. Specifically, a micron-order groove can be formed on the glass substrate by using the photolithography technique. However, since the production efficiency of a glass substrate is extremely low, including the time and effort required for surface processing, it is expected to be a serious problem in mass production.

On the other hand, a so-called resin chip using an organic polymer (resin) material in place of the glass substrate is more productive, less costly, or more waste than glass as a material for the above-mentioned medical diagnostic analysis device chip. It is expected that it will be superior in terms of processing, etc., and application of resin chips to small and simple analyzers is being studied. For example, an electrophoretic device composed of a capillary and a flat plate using a transparent resin (JP-A-2-259557), an organic polymer substrate for separation having a capillary-shaped trench (groove)
No. 327597), an electrophoretic analysis system using an acrylic resin chip (Anal. Chem., 6).
9, P.I. 2626-2630 (1997)) and the like.

However, since many biological substances to be measured by such an analyzer have only absorption of light in the ultraviolet region, that is, they are macroscopically colorless,
In order to use it for analysis for medical diagnosis, it is very important to be able to employ a measuring means using light having a wavelength in the ultraviolet region as a light source. For example, DNA with absorption at 260 nm
Can be directly detected without fluorescent labeling.

From these viewpoints, polymethylmethacrylate (PMM) used in these techniques is used.
A), polycarbonate (PC), polystyrene (P
S) or polyethylene terephthalate (PET)
Since general organic polymer materials such as have absorption in the ultraviolet region, when trying to detect the target substance with absorbance, which is a general detection method that has been used widely in the past, absorption by the chip material is extremely high. Large, resulting in inaccurate measurements. On the other hand, even in the case of fluorescent labeling, which is a detection method expected to have higher sensitivity, PMMA,
PS, PC, and PET have a problem in that the carbonyl group or aromatic ring contained therein absorbs ultraviolet rays and emits fluorescence, so that the detection sensitivity is greatly reduced in the background.

In addition, amorphous polymer materials such as PMMA and PC which have been conventionally used as the light transmitting resin are
There are problems with water absorption and degradability during injection molding. That is, from the viewpoint of thermal stability, for example, PMMA has a problem that a decomposition reaction based on depolymerization occurs at a relatively low temperature of 200 ° C., and the monomer remains in the molded body. Also, PC
Since is a condensation polymer, it has a drawback that hydrolysis due to residual water in the polymer may occur during molding. Furthermore, since resins such as PMMA, PC, PET are weak against polar solvents such as methanol and acetonitrile, μ-T
There is also a drawback that the solvent that can be used in AS is limited and the application is limited.

Attempts have been made to improve these drawbacks of conventional polymeric materials, and as novel polymeric materials, CHD-based monomers such as 1,3-cyclohexadiene (hereinafter referred to as CHD) or CHD derivatives, Further, cyclic polyolefins using dicyclopentadiene derivatives and various polycyclic norbornene-based monomers have been developed, and it is disclosed that these resins are excellent in transparency, heat resistance, chemical resistance, water resistance and the like. . Then, Japanese Patent Laid-Open No.
Japanese Patent Laid-Open No. 000-39420 discloses a resin microchip using these cyclic polyolefins as a substrate. As a method of forming microchannels,
A fine sample in which a liquid sample flows by ultrasonic fusion, heat fusion, adhesion with an adhesive such as a hot melt adhesive or UV adhesive, adhesion with an adhesive, pressure contact directly or through a thin elastic sheet, etc. A method of bonding a lower substrate having a flow channel and an upper substrate made of a flat film is disclosed. In addition, as a method for forming a substrate having a fine flow path through which a liquid sample flows, melt molding such as injection molding, compression molding, and extrusion molding is disclosed. Among them, injection molding, especially in the presence of carbon dioxide gas at a pressure of 10 MPa or less is present. It has been disclosed that injection molding in is preferred. However, the microchip of the above-mentioned invention has a problem in terms of adhesiveness between the lower substrate and the upper substrate and moldability of a fine channel.

[0011]

DISCLOSURE OF THE INVENTION The present invention is directed to a resin-made microchip having a flow path sealed by an upper substrate which is transparent, has a small absorption in an ultraviolet region, has a fine surface smoothness, and has a high dimensional accuracy. The challenge is to provide.

[0012]

In order to achieve the above-mentioned object, the present inventors have used a transparent hydrocarbon-based polymer while maintaining a fine flow path with good surface smoothness and high dimensional accuracy. The present invention has been completed by earnestly investigating a method of sealing a lower substrate with an upper substrate and a method of forming a lower substrate having a fine flow path with good surface smoothness and high dimensional accuracy.

That is, the present invention provides [1] a micro-sample for performing electrical and / or optical measurement of a liquid sample, which has at least a lower substrate having a fine channel through which the liquid sample flows and an upper substrate. In the chip, at least the lower substrate is made of a transparent hydrocarbon polymer represented by the following formula (1), and at least the surface of the lower substrate on the side having the flow channel is subjected to oxygen plasma treatment, A microchip obtained by thermocompression bonding the upper substrate at a temperature lower than the glass transition temperature (Tg) of the transparent hydrocarbon polymer,

[Chemical 2] [Formula (1) represents the composition formula of the polymer. (A)-(F)
Each represents a repeating unit constituting a polymer main chain. 1 to q each represent wt% of the repeating unit contained in the polymer main chain. (A): At least one repeating unit derived from a cyclic conjugated diene monomer. (B): At least one repeating unit derived from a cyclic olefin-based monomer. (C): At least one repeating unit derived from a vinyl aromatic monomer. (D): At least one repeating unit derived from a chain conjugated diene monomer. (E): At least one repeating unit derived from ethylene or an α-olefin monomer. (F): At least one repeating unit derived from other copolymerizable monomers. However, l + m + n + o + p + q = 100, 50 ≦ m +
p ≦ 100, the number average molecular weight of the polymer is 20,000 or more and 5
It is 00000 or less. ]

[2] The microchip according to the above [1], wherein the transparent hydrocarbon polymer represented by the formula (1) contains at least one cyclohexane ring in its main chain and / or side chain.

[3] In the formula (1), at least one selected from ethylene monomer, propylene monomer, ethylethylene monomer, isopropylethylene monomer and 2-ethylpropylene monomer as (E). The microchip according to the above [1] or [2], which contains a repeating unit derived from one kind, and the content of the repeating unit is 5 wt% or more and 70 wt% or less with respect to the total amount of the transparent hydrocarbon polymer,

[4] The microchip according to the above [1], [2] or [3], wherein the fine flow path through which the liquid sample flows is a flow path formed by laser ablation.

[5] A predetermined liquid sample flowing through the channel of the microchip, which is obtained by connecting the microchip according to [1], [2], [3] or [4] to a mass spectrometer. A device for analyzing components.

[0018]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention will be described in detail below.

The transparent hydrocarbon polymer in the present invention is
50 wt% or more in total of the repeating unit (B) derived from the cyclic olefin-based monomer and the repeating unit (E) derived from ethylene and the α-olefin-based monomer
A repeating unit (A) derived from a cyclic conjugated diene monomer, a repeating unit (C) derived from a vinyl aromatic monomer, and a chain conjugated diene monomer are required to be contained in an amount of 0 wt% or less. The repeating unit (D) derived from the above and other repeating units (F) derived from the copolymerizable monomer are contained as necessary.

The cyclic conjugated diene-based monomer in the present invention is a cyclic conjugated diene having a 5-membered ring or more composed of carbon-carbon bonds. A preferred cyclic conjugated diene-based monomer is a 5- to 8-membered cyclic conjugated diene composed of carbon-carbon bonds, and specific examples include 1,3-cyclopentadiene, 1,3-cyclohexadiene, 1, Examples thereof include 3-cyclooctadiene and derivatives thereof. From the viewpoint of stability such as heat resistance and weather resistance, a particularly preferred cyclic conjugated diene-based monomer is a 6-membered cyclic conjugated diene, and specific examples include 1,3-cyclohexadiene,
It is a 1,3-cyclohexadiene derivative. These monomers may be used alone or in combination of two or more.

The cyclic conjugated diene-based monomer in the present invention
As the repeating unit (A) derived from, the above cyclic conjugated di-
Repeating unit structure formed by polymerization of ene-based monomer
Examples of the repeating unit include a non-conjugated double bond.
The cyclic olefin-based monomer in the present invention means carbon-carbon.
Cyclic olefin with 5 or more membered rings composed of elementary bonds
And preferably a 5- to 8-membered cyclic olefin, particularly preferably
Preferred is a 6-membered cyclic olefin. As an example
Cyclopentene, cyclohexene, cyclooctene
And these derivatives can be illustrated. Also,
The cyclic olefin monomer has a norbornene structure
Including compounds having. As a specific example, bicyclo [2,2
2,1] hept-2-ene, tetracyclo [4,4,
0,1^2,5, 1^7,10] -3-dodecene, hexacyclo
[6, 6, 1, 1^3,6, 0^2,7, 0^9,14] -4-Heptade
Sen, tricyclo [5,2,1,0^2,6] -8-Decse
Pentacyclo [6,5,1,1^3,6, 0^2,7，
0^9,14] -4-Hexadecene, Pentacyclo [6,5,5]
1,1^3,6, 0^2,7, 0^9,13] -4-Pentadecene, Hep
Tacyclo [8,7,0,1^2,9, 1^4,7, 1^11,17，
0^3,8, 0 ^12,16] -5-Eicosane, Pentacyclo
[4,7,0,1^2,5, 0^8,13, 1^9,12] -3-Penta
Decene, tricyclo [4,4,0,1^2,5] -3-un
Decene, 5-carboxymethylbicyclo [2,2,1]
Hept-2-ene, 5-methyl-5-carboxymethyl
Bicyclo [2,2,1] hept-2-ene, 5-cyano
Bicyclo [2,2,1] hept-2-ene, 8-carbo
Xymethyltetracyclo [4,4,0,1^2,5，
1^7,10] -3-Dodecene, 8-carboxyethyl tetra
Cyclo [4,4,0,1^2,5, 1^7,10] -3-Dodese
8-carboxy n-propyl tetracyclo [4,
4,0,1^2,5, 1^7,10] -3-Dodecene, 8-carbo
Xysopropyltetracyclo [4,4,0,1^2,5，
1^7,10] -3-Dodecene, 8-carboxy n-butyl te
Tracyclo [4,4,0,1^2,5, 1^7,10] -3-Dode
Sen, 8-methyl-8-carboxymethyl tetracyclo
[4, 4, 0, 1 ^2,5, 1^7,10] -3-Dodecene, 8-
Methyl-8-carboxyethyltetracyclo [4,4,
0,1^2,5, 1^7,10] -3-Dodecene, 8-methyl-8
-Carboxy n-propyl tetracyclo [4,4,0,
1^2,5, 1^7,10] -3-Dodecene, 8-methyl-8-carb
Ruboxiisopropyltetracyclo [4,4,0,1]
^2,5, 1^7,10] -3-Dodecene, 8-methyl-8-cal
Boxy n-butyl tetracyclo [4,4,0,1^2,5，
1^7,10] -3-Dodecene, 8-ethylidene tetracyclo
[4, 4, 0, 1^2,5, 1^7,10] -3-Dodecene, 8-
Ethyl tetracyclo [4,4,0,1^2,5, 1^7,10]-
3-dodecene, 8-methyltetracyclo [4,4,0,
1^2,5, 1^7,10] -3-Dodecene, 6-ethyl-1,
4,5,8-Dimethano-1,4,4a, 5,6,7,
8,8a-octahydronaphthalene, 5,10-dimethyl
Rutetracyclo [4,4,0,1^2,5, 1^7,10] -3-
Dodecene, dimethanooctahydronaphthalene, ethyl te
Tracyclododecene, Trimethanooctahydronaphthale
Penta [8,4,0,1^2,5, 1^9,12, 0^8,13]-
3-hexadecene, penta [7,4,0,1^2,5, 1
^9,12, 0^8,13] -3-Pentadecene, Heptacyclo
[8, 7, 0, 1^3,6, 1^10,17, 1^12,15, 0^2,7, 0
^11,16] -4-Eicosane, heptacyclo [8,8,
0,1^4,7, 1^11,13, 1^13,16, 0^3,8, 0^12,17] -5
Mention may be made of Hen Eikosan and the like. These simple
The number of the monomers may be one or two or more, if desired.
Yes.

The repeating unit (B) derived from the cyclic olefin monomer in the present invention includes a repeating unit formed by polymerization of the above cyclic olefin monomer and the above cyclic conjugated diene monomer. The repeating unit obtained by hydrogenating the repeating unit (A) which originates is mentioned.

The vinyl aromatic monomer in the present invention is, for example, styrene, α-methylstyrene, o-methylstyrene, p-methylstyrene, p-tert-butylstyrene, 1,3-dimethylstyrene, 2 , 5-Dimethylstyrene, o-methoxystyrene, divinylbenzene, vinylnaphthalene, diphenylethylene, vinylpyridine, 1,2-dimethylstyrene, 1,1-diphenylethylene, 2,4-diphenyl-4-methyl-1-pentene , 1,3-diisopropenylbenzene and the like. These monomers may be used alone or in combination of two or more.

The repeating unit (C) derived from the vinyl aromatic monomer in the present invention is a repeating unit having an aromatic ring in the repeating unit structure, which is formed by polymerization of the vinyl aromatic monomer. Is mentioned. The chain conjugated diene-based monomer in the present invention is a hydrocarbon compound having 4 to 10 carbon atoms and having at least one conjugated diene structure. Specific examples include 1,3-butadiene, isoprene, 2,3-dimethyl-1,3-butadiene, 1,3-pentadiene, and 1,3-hexadiene. One of these monomers may be used alone, or two may be used.
It may be more than one kind.

The repeating unit (D) derived from the chain conjugated diene monomer in the present invention is a non-conjugated double bond in the repeating unit structure formed by polymerizing the above chain conjugated diene monomer. Examples include repeating units containing a bond. Ethylene in the present invention, the α- olefin-based monomer,
Examples thereof include ethylene, propylene, 1-butylene, 4-methylpentene, vinylcyclohexane and the like. These monomers may be used alone or in combination of two or more.

The repeating unit (E) derived from the ethylene or α-olefin-based monomer in the present invention is a repeating unit formed by polymerization of the above-mentioned ethylene or α-olefin-based monomer, and the above vinyl aroma. Examples thereof include a repeating unit (C) derived from a group-based monomer and a repeating unit obtained by hydrogenating a repeating unit (D) derived from a chain conjugated diene-based monomer.

The other copolymerizable monomer in the present invention is a conventionally known monomer which can be polymerized by anionic polymerization. Methyl methacrylate, methyl acrylate, acrylonitrile, methacrylonitrile, methyl vinyl ketone, polar vinyl monomers such as methyl α-cyanoacrylate, ethylene oxide, propylene oxide, cyclic lactone, polar monomers such as cyclic lactam, Other examples include alkylcyclosiloxanes such as hexamethylcyclotrisiloxane and octamethylcyclotetrasiloxane, and organic silicon compounds such as alkylarylsiloxanes. These monomers may be used alone or in combination of two or more. As the repeating unit (F) derived from the other copolymerizable monomer in the present invention,
The repeating unit formed by polymerizing the above-mentioned other copolymerizable monomer is included.

The transparent hydrocarbon polymer in the present invention is
Repeating unit (B) derived from a cyclic olefin-based monomer
And a repeating unit (E) derived from ethylene and an α-olefin-based monomer are contained in a total amount of 50 wt% or more (5
0 ≦ m + p ≦ 100). It is preferably 70 wt% or more, more preferably 90 wt% or more. Content rate is 50wt
%, Transparency, heat resistance, solvent resistance (here, solvent resistance to polar solvents used in μ-TAS, particularly water, acetonitrile, acetone, methanol, etc.) and hygroscopicity. It is preferable in terms of the above. The glass transition temperature (Tg), which is one of the indexes of heat resistance, is 80 ° C or higher, preferably 100 ° C or higher, more preferably 120 ° C or higher, still more preferably 150 ° C or higher, and particularly preferably 170 ° C. On the other hand, if the glass transition temperature is 400 ° C. or higher, the melt molding temperature becomes very high, and molding is difficult due to problems such as thermal decomposition, which is not preferable. In addition, the formula (1) of the present invention
When the transparent hydrocarbon-based polymer represented by is a block polymer and has a phase-separated structure, it may show two or more glass transition temperatures. The glass transition temperature here means the highest glass transition temperature. To tell. Further, the transparent hydrocarbon polymer in the present invention preferably contains at least one cyclohexane ring in its main chain and / or side chain.
By containing a cyclohexane ring, properties such as heat resistance and weather resistance are improved.

The structure of the transparent hydrocarbon polymer in the present invention may be either a block structure or a random structure. However, a random structure is preferable in terms of solubility in various solvents used for coating film formation and flow characteristics during melt molding. In particular, in the formula (1), a transparent hydrocarbon polymer in which the bonding structure of (B) is a random structure with (E) and satisfies 90 ≦ m + p ≦ 100 and 0.1 ≦ m / p ≦ 9, Solvent resistance, hygroscopicity, heat resistance, weather resistance, coating film formation,
It is preferable because it has excellent balance in terms of melt moldability. m /
When p is 0.1 or more, the composition ratio of the repeating unit derived from the cyclic olefin-based monomer is increased, and thus good heat resistance is exhibited. Further, when it is 9 or less, the bond between the repeating units derived from the cyclic olefin-based monomer has an appropriate length, and thus sufficient solvent solubility can be exhibited. 0.6 ≦ m from the balance of heat resistance and solvent solubility
More preferably, / p ≦ 5. In the formula (1), (E) is an ethylene monomer, a propylene monomer,
Ethyl ethylene monomer, isopropyl ethylene monomer,
And a repeating unit derived from at least one selected from 2-ethylpropylene monomer, and the content of the repeating unit is 5 wt% with respect to the total amount of the transparent hydrocarbon polymer.
It is preferable that the impact resistance is improved and the thermocompression bonding property after the oxygen plasma treatment is improved. If the content exceeds 70 wt%, the heat resistance is impaired, which is not preferable. Further, the repeating unit derived from at least one selected from ethylene monomer, propylene monomer, ethylethylene monomer, isopropylethylene monomer, and 2-ethylpropylene monomer is in a block state. Is preferred.

The number average molecular weight of the transparent hydrocarbon polymer in the present invention is 2 in terms of polystyrene equivalent average number molecular weight.
It is 0000 or more and 500,000 or less, and more preferably 30,000 or more and 200,000 or less. 200
When it is at least 00, strength and impact resistance as a polymer material can be exhibited. Further, when it is less than 500000, the solution viscosity when the solvent is dissolved can be appropriately adjusted.

The transparent hydrocarbon polymer in the present invention is
Light transmittance (ASTM D1003) is 85% or more, preferably 90% or more, and light wavelength region 240
Minimum value of light transmittance in the ultraviolet region of ~ 400nm is 1
It is 50% or more, preferably 70% or more, and more preferably 80% or more for a flat film having a thickness of 00 μm. When it is less than 50%, the detection sensitivity in the fluorescence labeling method is lowered due to the background of fluorescence due to ultraviolet absorption.

The liquid sample in the present invention may be a solution or a dispersion as long as it can smoothly flow through a fine channel (microchannel) in a microchip, and it may be a fluorescently labeled deoxyribonucleic acid ( DN
A), an aqueous dispersion solution of protein or the like, an organic (methanol, acetonitrile etc.) dispersion solution, a buffer solution of phosphoric acid, acetic acid or the like can be mentioned as an example.

As the optical measurement method in the present invention, an absorbance method (for example, N. Kuroda, et.a.
l. J. Chromatogr. , Vol. 798
P. 325-334 (1998)), fluorescence spectroscopy (for example, SC Jakobson, et. Al., Ana).
l. Chem. , Vol. 66 P. 4127-413
2 (1994), JP-A-2-245655), chemiluminescence method (for example, MF Regehr, et.a.
l. J. Capillary Electropho
r, Vol. 3 P. 117-124 (1996)) and the like. In the chemiluminescence method and the fluorescence method, the substance to be detected becomes a compound in an excited state by the reaction with an oxidant or the like and the excitation light, and from this state, the energy when the ground state is released is detected as light. Is the way to do it. On the other hand, the absorbance method is a method in which light is incident on a solution containing a substance to be detected, the intensity of transmitted light is measured, and the ratio of the intensity of transmitted light to the incident intensity is obtained. Generally, the sensitivity increases in the order of the absorbance method, the fluorescence method, and the chemiluminescence method.

As another measuring method, a photothermal conversion method (thermal lens method) in which a sample in a liquid is excited by excitation laser light to form a so-called thermal lens and a change in the thermal lens is detected by probe laser light ) Is mentioned. Detection of the probe laser light can be performed by a photodiode, a CCD camera, a photomultiplier tube, or the like.

The method of electrical measurement in the present invention can be performed by electrochemical measurement using electrodes, ion electrodes or the like, or by connecting a microchip to a gas chromatograph analyzer, a liquid chromatograph analyzer, or a mass spectrometer. Gas measurement,
Examples include liquid measurement and mass measurement (nanospray).

The shapes of the lower substrate and the upper substrate in the present invention are not particularly limited, but they are usually flat plates. The thickness of the lower substrate is usually 10 to 5000 μm. On the other hand, the thickness of the upper substrate is usually 1 to 500 μm, preferably 1
˜100 μm. Both the lower substrate and the upper substrate are preferably made of the transparent hydrocarbon polymer of the present invention. However, since the upper substrate is thinner than the lower substrate, a material other than a transparent hydrocarbon-based polymer, such as PET, PC, PMMA, or PS, is used as the upper substrate as long as the detection sensitivity is not significantly reduced by the background of fluorescence. It is also possible to do so.

The lower substrate in the present invention has a fine channel through which the liquid sample flows. The sectional shape of the fine channel is not particularly limited. It may have any shape such as a quadrangle, a trapezoid, a polygon such as a triangle, a semi-circle, a semi-oval, etc., but a quadrangle or a trapezoid is preferable. The maximum width is 1 to 5000μ
m, maximum depth 0.1-2000 μm, cross-sectional area 1-1
It is preferable that the thickness be 0,000,000 μm ² . More preferably, the maximum width is 2 to 500 μm, the maximum depth is 1 to 200 μm,
The cross-sectional area is ² to 100,000 μm ² .

The dimensional accuracy of the flow path is ± 5 with respect to the design dimension in order to obtain the accuracy of operation and the reproducibility between devices in the analysis and quantitative analysis of ultrafine material components. %, And the cross-sectional area is preferably within ± 7%. Also,
Width and depth are ± 2% for highly accurate quantitative analysis
Dimensional accuracy within ± 4% is more preferable.

On the other hand, the upper substrate according to the present invention may have a fine flow path through which the liquid sample flows on the surface facing the lower substrate, or may be a simple flat film.
The transparent hydrocarbon polymer in the present invention can be formed into a flat plate for a microchip by a known method. That is, injection molding, compression molding, extrusion molding or the like can be adopted.

The transparent hydrocarbon polymer of the present invention is
Dissolve in a soluble solvent and coat it on the substrate by an appropriate coating method such as spin coating, dip coating, blade coating, or roll coating, and then obtain a film by drying It can be processed into a flat film by a known cast film forming method. Is possible.
Preferred soluble solvents include cyclohexane, toluene, decalin, tetrahydrofuran, chloroform,
Examples thereof include cyclohexanone and 1,1-dimethoxycyclohexane, and these may be one kind or two or more kinds. By performing the cast film forming process from a soluble solvent, it is possible to obtain a flat film having an excellent balance of heat resistance, surface hardness, and impact resistance, and high surface smoothness.

In the present invention, as a method for forming a fine flow path, carbon dioxide 10 is added during the steps of injection molding, compression molding, extrusion molding, and filling the mold cavity with the lower substrate.
By forming a flow path at the same time when molding by a method such as injection molding after filling at a pressure of MPa or less, by embossing formed by pressing a mold with a fine flow path formed on a resin Examples thereof include a method, a method of forming by etching or photolithography, a method of forming by plasma treatment by covering with a mask, and a method of forming a flow path by using laser ablation. Among them, the method of forming using laser ablation is preferable because it can easily form a flow path having good dimensional accuracy and high surface smoothness. Also,
The method of forming a fine channel in the substrate using laser ablation is a preferable method because the electroosmotic flow can be controlled by changing the processing conditions.

As preferable conditions for laser ablation, wavelengths are 152 to 308 nm and 10 to 3000 mj.
/ Pulse, 50 Hz can be mentioned. The lower substrate is fixed to the X and Y stepper, and usually has a photomask and 1
Irradiate UV-laser pulse through 0: 1 lens. When forming the flow path, the lower substrate is normally moved in the horizontal direction at a speed of 0.15 mm / s or 0.2 mm / s. On the other hand, a liquid sample reservoir can be formed by irradiating a sufficient pulse without moving the flat membrane.

In the present invention, at least the surface of the lower substrate is treated with oxygen plasma, and then the lower substrate and the upper substrate are thermocompression-bonded at a temperature lower than the glass transition temperature of the transparent hydrocarbon polymer to prepare a microchip. . In the present invention, at least the surface of the lower substrate on the side where the fine flow paths are formed needs to be subjected to oxygen plasma treatment, but both the lower substrate and the upper substrate may be subjected to oxygen plasma treatment,
It is preferable because the adhesion becomes better and the electroosmotic flow can be made higher.

In the present invention, after the oxygen plasma treatment, the glass transition temperature (T
Thermocompression bonding at temperatures below g). The oxygen plasma treatment method uses a plasma stripper on the substrate surface, and normally pressure is 0.5 to 200 Pa and oxygen gas flow rate is 100 to 500 ml.
/ Min, power 100 to 500 W, time 1 to 1000 s
Process under the condition of ec.

Immediately thereafter, the glass transition temperature (Tg) of the transparent hydrocarbon polymer used for the substrate was 1 to 300 ° C., preferably 20 to 150 ° C. lower, and the speed was 0.1 to 1 m / m.
Thermocompression bonding using a laminator at min. When the transparent hydrocarbon-based polymer represented by the formula (1) of the present invention is a block polymer and has a phase-separated structure, it may show two or more Tg's. Says the transition temperature.

In the present invention, the liquid sample is driven by electroosmosis, electrokinetic phenomenon such as electrophoresis, capillary phenomenon, pressure and the like. It was found in the present invention that the oxygen plasma treatment significantly increases the electroosmotic flow. As a result of the surface analysis (X-ray photoelectron spectroscopy), the surface of the substrate is oxidized, the surface composition is greatly increased with the oxygen / carbon ratio of 10 to 30/70 to 90, and the modification of the surface improves the electroosmotic flow. It is thought that it contributes to. Usually, a method of coating the surface of the channel with another organic polymer having a polar group such as a sulfonic acid group or a carboxylic acid group to increase the electroosmotic flow is known, but in the case of the present invention, oxygen plasma is used. The treatment makes it possible to reduce the thermocompression bonding temperature of the substrate and improve the electroosmotic flow in one step. From the above viewpoint, the surface composition has an oxygen / carbon ratio of 1
It is preferably 0/90 or more, more preferably 20/80 or more.

Further, the oxygen plasma treatment of the present invention is carried out locally or partially by using a mask to improve the adhesion of the treated surface, and biochemical species such as protein and complementary deoxyribonucleic acid and photosensitivity. Chemical species such as compounds can be patterned on the surface of the substrate. for that reason,
A microchip for protein and deoxyribonucleic acid analysis and a microchip for converting light into an electric signal and detecting the electric signal can be prepared, and can be applied to various sensors and information storage devices.

The transparent hydrocarbon polymer used in the present invention may be obtained by any production method, and is not particularly limited, but the reaction temperature,
From the viewpoint of polymerization yield, catalyst cost, polymer molecular weight, molecular weight distribution, and primary structure control, industrially most preferable polymerization methods include the methods described below.

When a method of combining an organometallic compound containing a 1 (1A) group metal and an organic compound (complexing agent) capable of forming a complex with the organometallic compound as a polymerization catalyst, a transparent hydrocarbon is used. Since the polymerization of the system monomer proceeds like living polymerization, a transparent hydrocarbon polymer can be quantitatively obtained. Moreover, it is also possible to introduce other copolymerizable monomers.

Since the reaction proceeds in a living manner in this polymerization method, the molecular weight of the transparent hydrocarbon polymer can be freely controlled, and the molecular weight can be designed according to the purpose. Further, either a block copolymer or a random copolymer can be synthesized by selecting a polymerization catalyst. The polymerization catalyst used in the method for polymerizing the cyclic conjugated diene-based monomer is composed of an organometallic compound containing a 1 (1A) group metal and an organic compound (complexing agent) capable of forming a complex with the organometallic compound.

1 which can be used for the organometallic compound
The (1A) group metal is lithium, sodium, potassium, rubidium, cesium, or francium, and lithium and sodium can be exemplified as particularly preferable group 1 metals. These may be one kind or a mixture of two or more kinds as necessary.

In the method for polymerizing the cyclic conjugated diene-based monomer of the present invention, the organic lithium compound and the organic sodium compound which are particularly preferably used as the polymerization catalyst are one or at least one which is bonded to an organic molecule containing at least one carbon atom. It is a conventionally known compound containing two or more lithium atoms or sodium atoms. For example, methyllithium, ethyllithium, n-propyllithium, iso
-Propyllithium, n-butyllithium, sec-butyllithium, tert-butyllithium, pentyllithium, hexyllithium, allyllithium, cyclohexyllithium, phenyllithium, hexamethylenedilithium, 1,3-bis (1-lithium-1) , 3,3-
(Trimethyl-butyl) benzene, cyclopentadienyllithium, indenyllithium, butadienyldilithium, isoprenyldilithium, etc., or one of polymer chains such as polybutadienyllithium, polyisoprenyllithium, polystyryllithium, etc. A conventionally known organolithium such as an oligomer or polymer containing a lithium atom in its part, sodium naphthalene, α-methylstyrene sodium living tetramer, n-amyl sodium or the like, or polybutadienyl sodium, poly Examples of the conventionally known organic sodium such as an oligomer or a polymer containing a sodium atom in a part of a polymer chain such as isoprenyl sodium and polystyryl sodium can be exemplified.

Examples of particularly preferable organolithium compounds include n-butyllithium and cyclohexyllithium, and examples of the organosodium metal include sodium naphthalene and α-methylstyrene sodium living tetramer. The amount of the polymerization catalyst used in the present invention can be appropriately selected so as to obtain a desired molecular weight and a polymer structure.

As the organic compound (complexing agent) capable of forming a complex with an organometallic compound containing a Group 1 metal, amines, ethers and thioethers are preferable, and even one kind of these complexing agents is necessary. A mixture of two or more kinds may be used depending on the above. From the industrial viewpoint, ethers and amines are preferable as the complexing agent used in the present invention. Ethers include ethers containing one or more oxygen atoms in the molecule, for example, dimethyl ether, diethyl ether, isopropyl ether, tetrahydrofuran, tetrahydropyran, dimethoxyethylene glycol, diethoxyethylene glycol, dioxane, trioxane, 2,2-bis ether. (2-oxolanyl) propane, 1,
Examples thereof include 1-dimethoxycyclohexane and 1,1-diethoxycyclohexane. Of the amines, the tertiary (tertiary) amines are most preferred. Preferred tertiary amine compounds include trimethylamine, triethylamine, N, N-dimethylaniline, N, N,
N ', N'-tetramethyldiaminomethane, N, N,
N ', N'-tetramethylethylenediamine, N, N,
N ', N'-tetramethyl-1,3-propanediamine, tetramethyldiethylenetriamine, 1,8-diazabicyclo [5,4,0] undecene-7, diazabicyclo [2,2,2] octane, 4,4' -Bipyridyl and the like can be mentioned, but the most preferable tertiary amine compound which can be industrially adopted is tetramethylethylenediamine (TMEDA).

The method for preparing the polymerization catalyst comprising the above organometallic compound and the complexing agent is not particularly limited, and conventionally known techniques can be adopted as necessary. For example, an organometallic compound is dissolved in an organic solvent under an inert gas atmosphere, a solution of amines is added thereto and kept for a certain period of time, and then a monomer is added without isolating a polymerization catalyst (complex). It is also possible to carry out polymerization. The molar ratio of the organometallic compound to the complexing agent is in the range of M / N = 20/1 to 1/20, where M is the organometallic compound and N is the complexing agent, and the high molecular weight product has a high yield. To obtain

The polymerization method is not particularly limited, either.
Gas phase polymerization, bulk polymerization, solution polymerization or the like can be adopted. Polymerization solvents that can be used in the case of solution polymerization include aliphatic hydrocarbons such as n-butane, n-pentane, n-hexane and n-heptane, cyclopentane, methylcyclopentane, cyclohexane, methylcyclohexane, cycloheptane, Alicyclic hydrocarbons such as decalin,
Examples thereof include aromatic hydrocarbons such as benzene, toluene, xylene and ethylbenzene, and ethers such as diethyl ether and tetrahydrofuran. These polymerization solvents may be one kind or a mixture of two or more kinds.

The polymerization can be carried out at temperatures in the range of -100 to 120 ° C, most preferably 0 to 100 ° C.
Further, from an industrial point of view, it is advantageous to carry out at room temperature (20 ° C.) to 80 ° C. The polymerization system pressure is not particularly limited as long as it is within the above-mentioned polymerization temperature range and is within a pressure range sufficient to maintain the monomer and solvent in the liquid phase. The time required for the polymerization reaction cannot be particularly limited because it varies depending on the purpose or the polymerization conditions, but it is usually within 48 hours, and particularly preferably 1
~ 10 hours. The atmosphere of the polymerization system is preferably an atmosphere of dry nitrogen, argon, helium or the like inert gas.

The transparent hydrocarbon-based polymer is a non-conjugated double bond or vinyl compound contained in the repeating unit structure derived from the cyclic conjugated diene-based monomer, using a suitable hydrogenation catalyst after the completion of the polymerization reaction. An aromatic ring contained in the repeating unit structure derived from an aromatic monomer, and a non-conjugated double bond contained in the repeating unit structure derived from a chain conjugated diene monomer have a hydrogenated structure It is possible.

The hydrogenation rate of the non-conjugated double bond or aromatic ring is usually 90% or more, preferably 95% or more, more preferably 97% or more. If many non-conjugated double bonds and aromatic rings remain, the light transmittance in the ultraviolet region of the light wavelength region of 240 to 400 nm, the thermal stability, and the weather resistance are deteriorated, which is not preferable.

The transparent hydrocarbon polymer has a light transmittance,
In addition to the above hydrogenation, a conventionally known addition reaction to a carbon-carbon double bond can be carried out to introduce a substituent or a functional group as long as the physical properties such as heat stability and weather resistance are not adversely affected. Preferred substituents and functional groups include alkyl groups, aryl groups, silanol groups, hydroxyl groups, carboxylic acid groups, ester groups, acid anhydride groups, epoxy groups, carboxylate groups, amino groups, amide groups, imide groups, imino groups. , Oxazoline group, hydrazine group, hydrazide group, isocyano group, cyanato group, isocyanato group, silyl group, silyl ester group, silyl ether group, thiol group, sulfide group, thiocarboxylic acid group, and sulfonic acid group.

The hydrogenation reaction is carried out in the presence of the transparent hydrocarbon polymer and hydrogenation catalyst in a hydrogen atmosphere. A conventionally known method can be adopted as the reaction format. For example, a batch system, a semi-batch system, a continuous system, or a combination thereof can be adopted. As the hydrogenation reaction catalyst, both conventionally known homogeneous catalysts and heterogeneous catalysts can be used, and the kind and amount thereof are not particularly limited.

Industrially particularly preferred metals contained in the hydrogenation catalyst include titanium, cobalt, nickel, ruthenium, rhodium, palladium and the like. These metals may be used as a homogeneous catalyst in combination with an organic compound (ligand) having a functional group such as ether, amine, thiol, phosphine, carbonyl, olefin, diene, or activated carbon, alumina, barium sulfate. It may be used as a heterogeneous catalyst by supporting it on a carrier such as silica, titania, zeolite, or crosslinked polystyrene. From the industrial point of view, a heterogeneous catalyst capable of recovering the hydrogenation catalyst is preferable. In the present invention, the amount of the hydrogenation catalyst used in the hydrogenation reaction is appropriately selected depending on the hydrogenation conditions, but is generally in the range of 10 wtppm to 50 wt% as the metal concentration with respect to the polymer.

Preferred solvents for the hydrogenation reaction are aliphatic hydrocarbons such as n-butane, n-pentane, n-hexane and n-heptane, cyclopentane, methylcyclopentane, cyclohexane, methylcyclohexane, Examples thereof include cycloaliphatic hydrocarbons such as cycloheptane and decalin, aromatic hydrocarbons such as benzene, toluene, xylene and ethylbenzene, and ethers such as diethyl ether and tetrahydrofuran. These hydrogenation reaction solvents may be one kind or a mixture of two or more kinds.

The temperature of the hydrogenation reaction is in the range of 0 to 300 ° C, preferably 30 to 200 ° C, more preferably 90.
~ 200 ° C. The pressure of the hydrogenation reaction is usually 0.1 to
It is in the range of 25 MPa, particularly preferably 0.5 to 15
It is MPa. The time required for the hydrogenation reaction is not particularly limited because it depends on the concentration of the polymer solution, the temperature of the reaction system, and the pressure, but is usually 5 minutes to 240 hours.

As the method for separating and recovering the transparent hydrocarbon polymer of the present invention from the reaction solution, a method usually used when recovering the polymer from the reaction solution can be adopted. For example, a steam coagulation method of evaporating and removing the polymerization solvent by directly contacting the reaction solution with steam, a method of precipitating the polymer by adding it to a poor solvent of a polymer that is miscible with the polymerization solvent, and forming the reaction solution into a thin film. It is possible to exemplify a method of heating the above to distill off the solvent, a method of performing pelletization while distilling off the solvent with a vented extruder, etc., depending on the properties of the transparent hydrocarbon polymer and the solvent used. It is possible to adopt the optimum method.

When it is necessary to obtain a highly pure transparent hydrocarbon polymer in which the metal, amines, hydrogenation catalyst metal and the like contained in the polymerization initiator are extremely reduced, the transparent hydrocarbon polymer is preferably used. A method of solubilizing the metal ions in the combined solution with a suitable chelating agent and then extracting and removing by alternating current contact with high-purity ion-exchanged water, a method of removing ionic impurities using an ion-exchange resin column, carbon dioxide supercritical A method for removing metal ions and low molecular weight amines using the method can be carried out as necessary.

Further, when the transparent hydrocarbon polymer of the present invention is separated and recovered, the thermal stability of the copolymer, the stability against ultraviolet rays and the flame retardancy are improved, so that the physical properties such as light transmittance are adversely affected. Known stabilizers and antioxidants within the range of not exceeding, specifically phenol-based, organic phosphate-based, organic phosphite-based, amine-based, sulfur-based, silicon-containing, various stabilizers such as halogen-based, Antioxidant,
It is possible to add flame retardants. The general amount of these stabilizers, antioxidants, and flame retardants added is selected from the range of 0.001 to 5 parts by weight with respect to 100 parts by weight of the polymer.

[0068]

The present invention will be described in more detail below with reference to Examples and Comparative Examples, but the scope of the present invention is not limited to these Examples. The monomer, the hydrocarbon compound solvent and the ether compound used in the present invention were those which were previously purified by distillation under an inert gas. As the number average molecular weight and the weight average molecular weight distribution, values in terms of standard polystyrene measured by gel permeation chromatography (GPC) method are shown. The conversion rate of each monomer was determined by gas chromatography. The glass transition temperature (Tg) of the polymer was determined by a thermobalance. The hydrogenation rate was determined by nuclear magnetic resonance (NMR) analysis. In the solubility test, the solubility of various concentrations was visually determined in various solvents at room temperature.

Gas Chromatography Measurement A packed column having β, β'-oxydipropionitrile as a column packing, and a hydrogen flame ionization detector as a detector, GC-14 manufactured by Shimadzu Corporation is used. I was there. The mobile phase was He, the column temperature was 90 ° C, and the temperature of the injection and detector parts was 200 ° C. Ethylbenzene was used as an internal standard for obtaining the actual conversion rate of each component.

Gel permeation chromatography measurement A Showdex column (K-8) was used to measure the number average molecular weight and weight molecular weight distribution of the polymer after polymerization shown in the synthesis example.
02, 804, and 805) attached to Toso HLC-
An 8020 type device was used. The measurement was performed under the conditions of a tetrahydrofuran mobile phase and a column temperature of 40 ° C. The molecular weight was calculated as a polystyrene-equivalent molecular weight from a calibration curve prepared from the elution time of a standard polystyrene sample. Since the polymer after hydrogenation shown in the synthesis example has low solubility in tetrahydrofuran, the polymer was measured using PL-210 High Temperature GPC manufactured by Polymer Laboratory. The mobile phase was o-dichlorobenzene and the column temperature was 130 ° C.

Method for measuring glass transition temperature (Tg) The glass transition temperature was measured using a thermobalance (hereinafter referred to as DSC) (DSC-7 manufactured by Perkin Elmer Co., Ltd.) at a heating rate of 10 ° C. per minute.

Method for measuring hydrogenation rate NMR apparatus (JEOL-EX27 manufactured by JEOL Ltd.)
0) was used to determine the hydrogenation rate. Measurement solvent: o-dichlorobenzene -d _4, concentration: 12.5 mg / 0.5 c
m ³ , temperature: 135 ° C.

Surface analysis After UV-laser ablation, the surface after oxygen plasma treatment is subjected to X-ray photoelectron spectroscopy (hereinafter referred to as XPS).
(Kratos Axis Ultra X-ray
photoelectron spectrum
It was analyzed in r).

Electroosmotic Flow Measurement Using a high voltage generator (CZE1000, manufactured by Spellman), the method of Huang et al. (Anal. Che).
m. , 1988, 60, p. 1837-1838) to measure electroosmotic flow. The measurement conditions are phosphate buffer solution 13.4.
mM concentration, pH = 7, flow path length 2 cm, voltage 300 V.

Fluorescence Measurement Fluorescence Scanner (Molecular Dynamic)
(manufactured by s company) was used to measure the fluorescence. 450 as excitation light
530 nm fluorescence was measured using nm light. Further, as a liquid sample, a solution obtained by dissolving 114 mM of fluorescein (manufactured by Fluka) in 18 mM no phosphate buffer was used.

Electrophoresis analysis A high voltage generator (CZE1000, manufactured by Spellman) was used, and a CCD (charge transfer device) camera (CF 8) was used.
/ 4, manufactured by Kappa) confocal microscope (Axio
vert 25, Lamp HPO 100 / 2W Z
(manufactured by eiss). Detection data is IMAQ
Computerized on board. As reagents for electrophoretic analysis, fluorescein (manufactured by Fluka) and fluorescein biotin (manufactured by Sigma), fluorescein isocyanate dextran 4000
A conjugate (manufactured by Fluka) (70 μM) was used.

Synthesis Example 1 (CHD / α-MS / Bd copolymer) 5 dm ^{3 A} high pressure reactor was thoroughly dried with dry nitrogen and deoxidized. Then, 1755 g of cyclohexane and 21.3 g of tetrahydrofuran were added as a reaction solvent, and 148.5 g of 1,3-butadiene (hereinafter referred to as Bd) was further added, and the internal temperature was raised to 40 ° C. while stirring. Cyclohexane solution (hereinafter referred to as DiLi) of a 0.8 molar 1,3-bis (1-lithium-1,3,3-trimethylbutyl) benzene / triethylamine equimolar mixture 18.36
g was added to initiate polymerization. Polymerization temperature from 40 ℃ to 45 ℃
The stirring was continued for 1 hour. After 1 hour, cooling was started and the reactor was rapidly cooled to an internal temperature of 5 ° C. At this point, a part of the reaction solution was sampled and Bd was completely polymerized,
It was confirmed that the Bd monomer was consumed. Then, 608.6 g of absolutely dry tetrahydrofuran was added, and 110 g of α-methylstyrene (hereinafter, referred to as α-MS) and 341 g of cyclohexadiene (hereinafter, referred to as CHD) were sequentially added. The temperature during the reaction was controlled at 5 ° C to 15 ° C. After 12 hours, the reaction was stopped with 1.6 g of methanol.
The reaction solution was added to 20 times volume of methanol to carry out reprecipitation and recovery, and the polymer was obtained by drying under reduced pressure to obtain a glass transition temperature (T
g) was measured. Conversion rate of CHD is 93.1%, α-M
S was 83.3%. Tg was measured at two points and was -44 ° C on the low temperature side and 157 ° C on the high temperature side. The number average molecular weight was 68700.

Then, this polymer solution 1300 was dried under dry nitrogen.
g, and 5% palladium-alumina powder 55 manufactured by NE Chemcat Co., Ltd. dispersed in 1350 g of cyclohexane.
0 g (average particle size 40 μm) was mixed and added to a 5 dm ³ high pressure reactor. The inside gas was sufficiently replaced with high-purity nitrogen and then with high-purity hydrogen, and the inside of the reactor was adjusted to 7.85 MPa. Then, while maintaining the hydrogen pressure, the internal temperature of the reactor was gradually increased, and after reaching 160 ° C., the reaction was continued for 8 hours.
After cooling the internal temperature to room temperature, 0.2 μmPT under nitrogen
The palladium alumina powder catalyst was removed with a pressure filter using an FE membrane to obtain a transparent polymer solution. Pour the polymer solution after filtration into 4 times the total volume of acetone,
After precipitation, collection was performed. The recovered polymer is dried in a vacuum dryer to remove the residual solvent, CHD hydrogenation rate 98%, α-
A recovered polymer having an MS hydrogenation rate of 92% and a Bd hydrogenation rate of 100% was obtained.

Two glass transition temperatures were observed, indicating that the polymer after hydrogenation maintained the phase-separated structure. The glass transition temperature on the low temperature side was −44 ° C., and the glass transition temperature on the high temperature side was 209 ° C. 30 g of the hydrogenated polymer was dissolved in 70 g of toluene, coated on glass with a doctor blade, and dried at room temperature (25 ° C.) for 3 days. Then, it was peeled from the glass plate and dried in a vacuum dryer at 180 ° C. for 8 hours to give a thickness of 0.030 ± 0.
A flat film in the range of 002 mm (hereinafter referred to as a 0.03 mm flat film) and a flat film in the range of 0.10 ± 0.01 mm (hereinafter referred to as a 0.1 mm flat film) were obtained.

Synthetic Example 2 (Norbornene Ring-Opening Polymer) 30 g of an alternating copolymer of cyclopentane and ethylene (Zeonor 1600R manufactured by Nippon Zeon Co., Ltd.) which is a ring-opening polymer of norbornene which is a commercially available transparent hydrocarbon polymer. Was dissolved in 70 g of toluene, coated on glass with a doctor blade, and dried at room temperature (25 ° C.) for 3 days. After that, peel it off from the glass plate and dry it in a vacuum dryer at 160 ° C for 8 hours.
A flat membrane with a thickness in the range of 0.03 ± 0.002 mm (hereafter
It is called a flat film of 0.03 mm. ) And 0.100 ± 0.01
A flat film in the range of mm (hereinafter referred to as 0.1 mm flat film) was obtained.

Example 1 Using the flat film having a thickness of 0.1 mm obtained in Synthesis Example 1, a channel was first formed by UV-laser ablation to obtain a lower substrate. UV laser (Lambda P
hysik LPX 205i, made in Germany) and X, Y stepper (Microcontrol, made in France) were used. The flat film was fixed on X and Y steppers, and a UV-laser pulse of about 150 nm was irradiated at a condition of 200 mj / pulse and 50 Hz through a photomask and a 10: 1 lens. The flat membrane is 0.15 mm / s or 0.20 in the horizontal direction.
Moves at a speed of mm / s, length 40 mm, maximum width 40 μ
m and a depth of 40 μm, and a double T-shaped flow path was formed. The microscope picture of the flow path of the lower substrate formed in FIG. 1 is shown. A rectangular flow path with high surface smoothness was formed by laser ablation. On the other hand, the 0.03 mm-thick flat film obtained in Synthesis Example 1 was used as the upper substrate, and the lower substrate and the upper substrate were combined so as to be a transparent hydrocarbon polymer having the same composition. Plasma stripper (MICRO-ONDES)
TEPLA300), pressure 101 Pa, oxygen gas flow rate 400 ml / min, power 500 W, time 15 sec
Oxygen plasma treatment was performed under the condition of c. Temperature 1 soon after
Laminator (RLM4 at 30 ° C, speed 0.2m / min)
19P, manufactured by Switzerland), and the two substrates were thermocompression bonded so that the surfaces subjected to the oxygen plasma treatment were aligned with each other. It was confirmed by observing the cross section of the flow channel with a microscope that good adhesion can be achieved at a temperature of several tens of degrees Celsius lower than Tg. 2A to 2C show micrographs of the cross section of the flow path after thermocompression bonding of the upper substrate. PE
It was found that an extremely clean flow path was formed without the protrusion of the melted adhesion layer because the adhesion layer was not used. The surface before and after the oxygen plasma treatment is X-ray photoelectron spectroscopy (hereinafter referred to as XPS) (Ult manufactured by Kratos Axis).
ra), the oxygen / carbon ratio was 9.7 / 96.
This was a large increase from 9 to 23.8 / 76.2, confirming surface modification.

Example 2 Using the microchip obtained in Example 1, electroosmotic flow was measured. Electroosmotic flow is 6.56 × 10 ^-8 m ² s ^-1 V
^−1, and an electroosmotic flow of a microchip produced by using a PE adhesion layer in Comparative Example 2 to be described later 5.22 × 10 ⁻⁸ m
^The value was higher than ² s ^-1 V ^-1 . It is considered that this is because the electroosmotic flow is high due to the modification of the microchip surface by the oxygen plasma treatment.

Example 3 The results of the fluorescence measurement of the microchip obtained in Example 1 are shown in FIG. 3B. The results of Comparative Example 1 are shown in FIG. The microchip prepared by using PET and the PE adhesion layer in Comparative Example 1 looks black in the background due to the fluorescence caused by PET, whereas the microchip obtained in Example 1 looks white. From this, it can be seen that there is almost no background due to fluorescence. As a result, the fluorescent material can be analyzed with higher sensitivity.

Example 4 The results of electrophoretic analysis of the microchip obtained in Example 1 are shown in FIG. Fluorescein (manufactured by Fluka) and fluorescein biotin (manufactured by Sigma) were used as test samples. A 10 mM phosphate buffer was used as a buffer. In addition, a 230 μm double T-shaped flow path was adopted. As a result of electrophoresis, it was confirmed that the above two components could be separated very well and could be separated in an extremely short time.

Example 5 Thickness 0.1 mm and thickness 0.03 mm obtained in Synthesis Example 2
The electroosmotic flow was measured using a microchip prepared by the same method as in Example 1 using the flat membrane of Example 1. The electroosmotic flow was 6.32 × 10 ⁻⁸ m ² s ⁻¹ V ^−1, and the electroosmotic flow of the microchip produced using the PE adhesion layer in Comparative Example 2 described later was 4.72 × 10 ^−8. m ² s ^-1 V ^- were higher than ^1. It is considered that this is because the electroosmotic flow is increased due to the effect of modifying the surface of the microchip by the oxygen plasma treatment.

Example 6 A microchip was prepared in the same manner as in Example 1 except that the flat film having a thickness of 0.1 mm and 0.03 mm obtained in Synthesis Example 1 was used and the electric power was set to 200 W under plasma processing conditions. Created. 4. The electroosmotic flow was measured in the same manner as in Example 2, and 5.
A value of 50 × 10 ^-8 m ² s ^-1 V ^-1 was obtained.

Example 7 A microchip was prepared in the same manner as in Example 5 except that the flat film having a thickness of 0.1 mm and 0.03 mm obtained in Synthesis Example 2 was used and the electric power was 200 W under the plasma processing conditions. Created. 4. The electroosmotic flow was measured in the same manner as in Example 2, and 5.
A value of 28 × 10 ^-8 m ² s ^-1 V ^-1 was obtained.

Example 8 Using the 0.1 mm thick flat membranes obtained in Synthesis Examples 1 and 2,
A flow path was formed by the same method as in Example 1 to obtain a lower substrate. This lower substrate was subjected to oxygen plasma treatment under the same conditions as in Example 1, and a 0.03 mm-thick PET flat film was used as the upper substrate, and a 0.03-mm-thick PE flat film was used as the adhesion layer. A microchip was obtained by thermocompression bonding using a laminator at 125 ° C. and a speed of 0.2 m / min. As a result of measuring the electroosmotic flow, the microchip prepared from Synthesis Example 1 was 5.98 × 10 ⁻⁸ m ² s ⁻¹ V ⁻¹ and the microchip prepared from Synthesis Example 2 was 5.60 × 10 ⁻⁸ m. ² s ^-1 V ^-1
The value of was shown.

Example 9 A microchip was prepared by the same method as in Example 1 and the variation between lots was observed. The results are shown in Table 1. It can be seen that there is almost no variation in the time required to transfer a liquid sample having a channel length of 2 cm, and reproducibility is high.

Example 10 The microchip obtained in Example 1 was used as a mass spectrometer (Fi
A sample of 1 mg / ml myoglobin was measured by a device (nanospray) connected to Nnigan. The result is shown in FIG. The peak intensities of all signals are as high as 10 ⁸ (the background is 10 ⁵ from the measurement result of the sample to which myoglobin is not added, as shown in FIG. 6), and stable measurement results are obtained.

Comparative Example 1 Using PET with a thickness of 0.1 mm, a flow path was formed by the same method as in Example 1 to obtain a lower substrate. Using a 0.03 mm thick PET film as the lower substrate and the upper substrate and a 0.03 mm thick PE film as the adhesion layer, thermocompression bonding was performed using a laminator at a temperature of 125 ° C. and a speed of 0.2 m / min. Got a chip. The photograph which observed the cross section of the flow path with the microscope is shown in FIG. 2D. Although the adhesion was achieved, the protrusion of the melted adhesion layer in the channel was observed.
Further, the result of fluorescence measurement performed by the same method as in Example 3 is shown in FIG. 3A.

Comparative Example 2 Using 0.1 mm thick flat membranes obtained in Synthesis Examples 1 and 2,
A flow path was formed by the same method as in Example 1 to obtain a lower substrate. Using a PET flat film having a thickness of 0.03 mm as the lower substrate and the upper substrate and a PE flat film having a thickness of 0.03 mm as the adhesion layer, thermocompression bonding was performed in the same manner as in Comparative Example 1 to obtain a microchip. As a result of measuring the electroosmotic flow, the microchip prepared from Synthesis Example 1 was 5.22 × 10 ^-8 m ² s ^-1 V
^-1 is 4.72 × for the microchip prepared from Synthesis Example 2.
The value was 10 ^-8 m ² s ^-1 V ^-1 .

[0093]

[Table 1]

[0094]

Industrial Applicability According to the present invention, it is possible to provide a resin-made microchip which is transparent, has a small absorption in the ultraviolet region, has a fine flow path having a good surface smoothness and a high dimensional accuracy. Moreover, since the electroosmotic flow can be increased by the oxygen plasma treatment, the liquid sample can be quickly transferred.

[Brief description of drawings]

FIG. 1 is a photomicrograph from the upper right of a channel formed by UV-laser ablation of Example 1.

2A is a micrograph of a cross section of a channel formed in a lower substrate of Example 1, and FIG. 2B is a cross section of a channel formed by thermocompression bonding after the oxygen plasma treatment of Example 1. A micrograph, (C) an enlarged micrograph of (A), and (D) a micrograph of a cross section of a channel formed by thermocompression bonding with a PET flat film as an upper substrate using the PE adhesion layer of Comparative Example 1. is there.

FIG. 3 is a photomicrograph of fluorescence measurement of the microchips obtained in Examples 1 and 2 and Comparative Example 1. (A)
Comparative Example 1, (B) Example 1.

FIG. 4 is a graph diagram of an electrophoretic analysis of the microchip obtained in Example 1.

5 is a graph showing measurement results of 1 mg / ml myoglobin of the microchip mass spectrometer (nanospray) obtained in Example 10. FIG.

FIG. 6 is a graph showing the measurement results (background) of a sample of the microchip mass spectrometer (nanospray) obtained in Example 10 in which myoglobin is not added.

Front Page Continuation (51) Int.Cl. ⁷ Identification code FI Theme Coat (reference) G01N 27/26 331K (72) Inventor Hubert Et Jiro, CH-888 Switzerland, Switzerland (72) Inventor Natori Shizuoka prefecture Fuji Sameshima second address of 1 Asahi Kasei Co., Ltd. in the F-term (reference) 4F073 AA01 BA06 BA07 BB02 BB03 CA01 HA11 4J100 AA02Q AA03Q AA04Q AA17Q AA20Q AB01S AB02S AB03S AB04S AB07S AB08S AB13S AB16S AQ12S AR04P AR05P AR09P AR10P AR11P AR15R AR17R AR18R AS01T AS02T AS03T AS04T BA05S BC43S BC69S CA03 CA04 CA05 CA06 DA36 DA62 DA65 HE29 JA17 JA24

Claims (5)

[Claims] 1. A microchip for performing electrical and / or optical measurement of a liquid sample, the microchip having at least a lower substrate having a fine channel through which a liquid sample flows and an upper substrate. The substrate is made of a transparent hydrocarbon-based polymer represented by the following formula (1), and at least the surface of the lower substrate on the side having the flow path is subjected to oxygen plasma treatment. A microchip obtained by thermocompression bonding at a temperature lower than the glass transition temperature (Tg) of a polymer. [Chemical 1] [Formula (1) represents the composition formula of the polymer. (A)-(F)
Each represents a repeating unit constituting a polymer main chain. 1 to q each represent wt% of the repeating unit contained in the polymer main chain. (A): At least one repeating unit derived from a cyclic conjugated diene monomer. (B): At least one repeating unit derived from a cyclic olefin-based monomer. (C): At least one repeating unit derived from a vinyl aromatic monomer. (D): At least one repeating unit derived from a chain conjugated diene monomer. (E): At least one repeating unit derived from ethylene or an α-olefin monomer. (F): At least one repeating unit derived from other copolymerizable monomers. However, l + m + n + o + p + q = 100, 50 ≦ m +
p ≦ 100, the number average molecular weight of the polymer is 20,000 or more and 5
It is 00000 or less. ] 2. The microchip according to claim 1, wherein the transparent hydrocarbon polymer represented by the formula (1) contains at least one cyclohexane ring in its main chain and / or side chain. 3. In the formula (1), an ethylene monomer, a propylene monomer, an ethylethylene monomer as (E),
It contains a repeating unit derived from at least one selected from isopropylethylene monomer and 2-ethylpropylene monomer, and the content of the repeating unit is 5 wt% or more and 70 wt% or less with respect to the total amount of the transparent hydrocarbon polymer. The microchip according to claim 1 or 2, which is 4. The microchip according to claim 1, 2 or 3, wherein the fine channel through which the liquid sample flows is a channel formed by laser ablation. 5. An apparatus for analyzing a predetermined component of a liquid sample flowing through a channel of the microchip, which is obtained by connecting the microchip according to claim 1, 2, 3 or 4 to a mass spectrometer.