JP2002027984A - Microreactor chip, method for testing chemical reaction, and thin film material for microreator chip - Google Patents

Abstract

PROBLEM TO BE SOLVED: To simplify production work to enhance the productivity, and to enhance the production accuracy for a microreactor chip, for a method for testing a chemical reaction, and for a thin film material for the microreactor chip. SOLUTION: This microreactor chip has a chip substrate 1 and a thin film material 2 laminated on the chip substrate 1, wherein the thin film material 2 has an opening 3 for containing a specimen in cooperation with the chip substrate 1 in the laminated state of the thin film material 2 on the chip substrate 1.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a microreactor chip, a method for testing a chemical reaction, and a thin film member for a microreactor chip for causing a predetermined chemical reaction with a small amount of sample.

[0002]

2. Description of the Related Art Conventionally, a chip (microreactor chip) for detecting a biological sample or performing chemical synthesis.
Have been developed. Hereinafter, each conventional technique in such a chip for biological sample detection and a chip for chemical synthesis will be described. First, a chip for detecting a biological sample will be described. As a chip for detecting a biological sample, for example, D
There are a chip (DNA chip) used for analyzing the base sequence of NA (deoxyribonucleotide) and a diagnostic chip utilizing an antigen-antibody reaction or an enzyme reaction.

[0003] DNA is composed of four types of bases, and the bases to which these bases bind are determined. In the hybridization method, such a base sequence is analyzed by utilizing this principle. That is, a DNA whose base sequence to be tested is unknown is mixed with a plurality of types of DNAs whose base sequences to be compared are known, and the DNA to be tested is mixed.
By investigating whether or not the NA and the DNA to be compared are bound, the DNA that can bind to the DNA to be tested is specified. This makes it possible to determine that the sequence complementary to the specified base sequence of the DNA is the base sequence of the test target DNA or a part of the base sequence of the test target DNA.

[0004] Specifically, a plurality of types of D to be compared are compared.
A solution containing NA (probe DNA) (hereinafter, also referred to as a probe solution) is attached to a slide glass in a dot shape (this is called spotting) and fixed. After a solution containing the DNA to be tested is dropped onto a slide glass (= DNA chip) to which the probe DNA is attached, hybridization is performed under predetermined conditions under which a DNA binding reaction can occur. The DNA chip is then washed, and the DNA solution that has not bound to the probe DNA is washed away. Since the unknown DNA to be inspected is fluorescently labeled in advance, only the portion where the DNA to be inspected and the probe DNA to be compared are bound becomes fluorescently labeled.

[0005] Therefore, by scanning the DNA chip with laser light and measuring the amount of fluorescence at each position on the slide glass, it is possible to quickly determine whether or not each probe DNA has bound to the DNA to be tested. The base sequence of the DNA to be tested can be analyzed based on the determination result. A DNA chip used for such a hybridization method has a predetermined size (for example, 25 mm × 7 mm).
5mm) on a chip substrate (eg, slide glass)
It is created by spotting a large number (tens to tens of thousands) of probe DNAs.

In order to efficiently perform the hybridization method, it is necessary to increase the spot density (the number of probe DNA spotted per unit area) and increase the number of probe DNAs on a chip substrate (DNA chip). . In a commercially available general DNA chip, a single silica coat is applied to the surface of the chip substrate,
The number of spots on the substrate is increased by adjusting the degree of spread of the droplet by the surface tension of the silica coat.

However, simply applying a single coating process to the surface of the chip substrate as described above requires only the probe D
It is not possible to sufficiently restrict the NA from spreading around, and the size of the probe DNA per spot (hereinafter, referred to as spot area) becomes relatively large. For this reason, there is a limit in increasing the spot density by reducing the distance between adjacent probe DNAs, and a further increase in the spot density is required in order to increase the efficiency of the analysis operation.

[0008] As a method for increasing the spot density,
It is known to form a frame for accommodating probe DNA using a photosensitive resin on the surface of a chip substrate such as glass. Since the spot area can be regulated by accommodating the probe DNA in the frame, the distance between adjacent probe DNAs can be reduced, and the spot density can be increased.

As a technique for forming a frame on such a chip substrate, Japanese Patent Application Laid-Open No. 11-187900 discloses a technique in which a matrix pattern having a frame structure having recesses arranged in a matrix is formed on a solid layer. A technique for forming a matrix pattern is disclosed. As a technique for forming a matrix pattern, there is a method in which a photoresist is coated on a resin coated on a chip substrate surface, and after patterning, the resin is patterned by a process such as etching. A method of coating a photosensitive resin on the surface of a chip substrate, exposing and developing the photosensitive resin itself by a photolithography process using a photomask, and, if necessary, further curing and patterning. ing.

Next, a microreactor chip for chemical synthesis will be described. In chemical synthesis, a method of containing a predetermined sample in a container such as a test tube or a beaker and mixing the sample in the container to perform a predetermined synthesis reaction is generally performed. Research for chemical synthesis using a microreactor chip has been conducted for the purpose of realization.

As one example of such a microreactor chip for chemical synthesis, a plurality of microchannels for flowing a plurality of types of samples are formed on a chip substrate, respectively, and these microchannels are formed on the chip substrate. Some are formed so as to merge on a substrate. in this case,
Since a predetermined synthesis reaction is performed in the merged flow channel only by flowing a small amount of a plurality of types of samples into the micro flow channel, the synthesis reaction proceeds while the samples flow through the flow channel. It is possible to observe the progress of the synthesis reaction.

[0012] Such a microreactor chip for chemical synthesis can perform a synthesis reaction with a very small amount of sample (reaction solution), so that the synthesis reaction can be performed efficiently.
When only a small amount of sample is available, as in biochemical reactions,
It is valid. In addition, the same kind of reaction can be performed in a large amount as in screening using a synthetic robot or the like, which is efficient. Furthermore, in recent years, it has attracted particular attention because of its ability to safely carry out a synthetic reaction accompanied by a violent exothermic reaction and its easy handling.

As such a microreactor chip, for example, JP-A-10-337173 discloses a microreactor chip in which a plurality of independent reaction chambers are formed on the surface of a silicon substrate (chip substrate). The reaction chamber is composed of a plurality of inlet ports, outlet ports, and a channel connecting these inlet ports and outlet ports.
It is formed on the surface of a silicon substrate (chip substrate) by anisotropic etching.

[0014]

However, in the above-mentioned prior art, in the case of a DNA chip, a photoresist or a photolithography technique is used to form a frame structure on the surface of a chip substrate. For this purpose, it is necessary to coat the surface of the chip substrate with a resin, and further, it is necessary to perform patterning and etching on the coated resin, which poses a problem that the production is extremely troublesome.

[0015] Similarly, a chip for chemical synthesis also has a problem in that a microchannel is formed on the surface of a silicon substrate (chip substrate) by anisotropic etching, so that it takes a very long time to manufacture. Further, providing a micro flow path on the surface of the chip substrate, that is, forming a groove in the chip substrate, has a problem that such groove processing has relatively low processing accuracy. That is, in the groove processing, a slight difference may occur in the depth and edge shape of the groove depending on the material of the chip substrate and the processing conditions.

SUMMARY OF THE INVENTION The present invention has been made in view of the above-described problems, and has been made in consideration of the above-described problems, and has been made in view of the above-mentioned circumstances, and has been made in view of the above-mentioned circumstances. An object of the present invention is to provide a chemical reaction test method and a thin film member for a microreactor chip.

[0017]

According to the first aspect of the present invention, there is provided a microreactor chip comprising: a chip substrate;
An opening for accommodating a sample in cooperation with the chip substrate in a state in which the thin film member is laminated on the chip substrate; It is characterized by having.

In this case, the thickness of the thin film member is 3 μm to 5 μm.
It is preferably in the range of 00 μm (claim 2). Further, the opening provided in the thin film member may be constituted by a plurality of independent fine holes. in this case,
The pore size of the micropores is preferably in the range of 10 μm to 500 μm (claim 4).

Alternatively, the opening provided in the thin film member may be formed as a flow path for flowing the sample. In this case, the width dimension of the flow path is 10 μm.
It is preferably in the range of m to 1000 μm (claim 6). Preferably, the sample is an oligonucleotide or a peptide (claim 7).

According to a eighth aspect of the present invention, there is provided a chemical reaction test method, wherein a chemical reaction test is performed using the microreactor chip according to any one of the first to seventh aspects. According to a ninth aspect of the present invention, there is provided the thin film member for a microreactor chip, wherein an opening portion used for accommodating a sample is provided, divided and laminated on the chip substrate.

[0021]

Embodiments of the present invention will be described below with reference to the drawings. (A) Description of First Embodiment First, in a first embodiment of the present invention, an opening provided in a thin film member is constituted by a plurality of independent small holes. A DNA chip (microreactor chip), a hybridization method (chemical reaction test method) using the DNA chip, and a thin film member used for the DNA chip. Hereinafter, these will be described. 1 to 5 are views showing a DNA chip of the present embodiment.

The DNA chip of the present invention is shown in FIG.
As shown in FIG. 1B, the semiconductor device includes a chip substrate 1 and a hydrophobic thin film member 2 that is stacked and attached to the chip substrate 1. The thin film member 2 has a plurality of (for example, 1,000 to 80,000) minute holes (opening portions) 3.
Is formed in a concave portion 3a formed from the opening 3 and the surface of the chip substrate 1, and a DN as a biological sample is provided.
A water-soluble probe solution (probe DNA) containing A is accommodated.

Hereinafter, the chip substrate 1, the thin film member 2 and the opening 3 will be described. First, the chip substrate 1 will be described. Here, the size of the chip substrate 1 is approximately the same as that of a generally used slide glass (for example, 25 mm).
The thickness is set to about 1 mm (about 0.7 mm to 1.6 mm), although it depends on the setting of an apparatus for detecting DNA and the like. Further, as described above, the concave portion 3a for accommodating the probe solution is formed by the surface of the chip substrate 1 and the opening 3 of the thin film member 2, and the surface of the chip substrate 1 on the side for accommodating the probe solution is formed. Preferably, a hydrophilic treatment is performed. Here, as such a hydrophilic treatment, for example, a treatment is performed such that a coating agent for immobilizing DNA in the probe solution is applied to form the coating film 1a as described later.

The material of the chip substrate 1 may be glass or resin. When using resin, this resin (base resin)
May be thermoplastic, thermosetting, or radical curable. Further, any of a homopolymer, a copolymer, a block polymer, and a graft polymer may be used. Those having excellent optical properties and exhibiting almost no absorption in the wavelength region of 400 nm or more are preferable, and those having no background noise upon fluorescence detection are particularly preferable. For example, acrylic resin such as polymethyl methacrylate and its copolymer, polystyrene or its copolymer, MS resin (random copolymer of methyl methacrylate and styrene), polycarbonate, polyalkylene terephthalate, aliphatic or alicyclic Formula polyamide, transparent polyolefin (polymethylpentene, polyethylene (co)
Polymers, polypropylene (co) polymers, etc.), various polymers derived from cycloolefins or cycloalkanes, polyvinyl acetate, polyvinylpyrrolidone, AS
Resin and transparent such as SAN resin (copolymer of acrylonitrile and styrene), ABS resin (acrylonitrile-butadiene-styrene resin), polyvinyl chloride, polyvinyl fluoride, polyvinylidene fluoride, polyarylate, polysulfone, polyethersulfone, etc. Cured resin derived from a thermoplastic resin having properties, triacetyl cellulose or a partially saponified product thereof, a compound having a functional group having radical polymerizability or thermopolymerizability (used for lenses, optical disks, optical transparent parts, etc. Various cured products), various rubbers and elastomers having transparency, and the like, but are not limited thereto.

Among them, acrylic resin such as polymethyl methacrylate and its copolymer, styrene resin such as MS resin, polycarbonate, transparent polyolefin (polyethylene and polypropylene), alicyclic olefin and cycloalkane derivative Thermoplastic resins such as various resins having transparency derived from the above are preferred. When such a resin is used, the chip substrate 1 is molded by a usual molding method according to the characteristics of each resin. For example, molding by injection molding such as injection molding, extrusion molding, compression molding, injection compression molding, transfer molding, calendar molding, or cast molding can be exemplified, but is not limited thereto.

As described above, the surface of the chip substrate 1 on the side for containing the probe solution is provided with the D in the probe solution.
It is preferable that a coating agent for immobilizing NA is applied, and as such a coating agent,
(1) a coating agent having a positive charge as a surface functional group, which is suitable for immobilizing DNA by ionic bonding;
(2) a coating agent having a surface functional group suitable for immobilizing DNA by hydrogen bonding, or (3) (modification,
(Oligo) A coating agent having, as a surface functional group, a functional group suitable for immobilizing a DNA terminal amino group by covalent bond is preferable.

Typical examples of the functional group (1) include a quaternary amino group (ammonium group), a phosphonium group, a sulfonium group, a biguanide group, and a betaine group (typically, a quaternary amino group and a COO group). And a quaternary amino group is particularly preferable. Representative examples of the functional group (2) include a urethane group, a urea group, a hydrazide group, an amide group,
Examples thereof include an amino group, a hydroxyl group, a carboxylic acid group, a sulfonic acid group, and a phosphoric acid group. In view of easiness of introduction, durability of coating, effects, and the like, a urethane group, a hydroxyl group, and the like are particularly preferable.

Typical examples of the functional group (3) include a ketone group, a carbonyl group which forms a Schiff base with an amino group such as an aldehyde group, an epoxy group which is a group which undergoes an addition reaction with the amino group, Such as an azlactone group or an episulfide group, and an acryloyl group, a methacryloyl group, an acrylamide group, a methacrylamide group, or a maleimide group, which forms a covalent bond by performing an addition reaction such as Michael addition with an amino group. Thinking,
Particularly preferred are carbonyl, epoxy, acryloyl and methacryloyl groups.

The coating method of such a coating agent is as follows.
Typically, a substrate is coated with a coating device such as a dip coating method, a spin coating method, a flow coating method, a spray coating method, a bar coating method, and a gravure coating, a roll coating, a blade coating, and an air knife coating. The coating is performed by drying the solvent (and irradiating with active energy rays, if necessary) to obtain a smooth coating film of 0.1 μm to 50 μm, preferably 0.2 μm to 5 μm on the surface of the substrate.

When active energy ray curing is required,
In order to cross-link and cure the applied coating composition layer, ultraviolet light emitted from a light source such as a xenon lamp, a low-pressure mercury lamp, a high-pressure mercury lamp, an ultra-high-pressure mercury lamp, a metal halide lamp, a carbon arc lamp, a tungsten lamp, or an electron beam of usually 20 to 2,000 kV An active energy ray such as an electron beam, α-ray, β-ray, or γ-ray taken out of the accelerator is irradiated and cured to form a coating film.

Next, the thin film member 2 and the opening 3 will be described. The thin film member 2 is made of resin and has hydrophobicity. The resin used may be a thermoplastic resin or a thermosetting resin. In particular, a polyolefin resin such as polyethylene or polypropylene, an acrylic resin such as polymethyl methacrylate, a polystyrene resin, or a polycarbonate resin may be used. Such a thermoplastic resin is preferred. These may be homopolymers, copolymers, block polymers, and graft polymers.

The thickness T1 of the thin film member 2 is 3 μm to 3 μm.
It is set in the range of 500 μm. Thickness T1 of thin film member 2
Is the depth of the recess 3a for accommodating the sample (probe solution) formed by the opening 3 of the thin film member 2 and the chip substrate 1, and the depth is set within the above range. Thus, the sample can be stably accommodated, and the amount of the accommodated sample can be minimized.

The resin molding method is the same as the molding method applied when a resin is used as the material of the chip substrate 1. The opening 3 is formed by penetrating the thin film member 2 having a predetermined thickness by laser processing or punching. Here, each opening 3 is formed in a perfect circular shape, and the diameter (opening dimension) D1 is not particularly limited as long as it is 10 μm or more, but is preferably set in the range of 10 μm to 500 μm. Accordingly, spotting into the opening 3 becomes relatively easy, and the number of spots of the probe solution on the chip substrate 1 can be sufficiently secured.

The shape of the opening 3 is not limited to a perfect circle, and may be, for example, an ellipse, a quadrangle, or another polygon. However, a perfect circle is preferable in terms of ease of processing. In the case of an ellipse, it is preferable to set the length of the major axis in the range of 10 μm to 500 μm.
It is preferable to set in the range of 0 μm to 500 μm.
That is, it is preferable to set the maximum dimension (opening dimension) in the shape of the opening 3 in plan view in the range of 10 μm to 500 μm.

Further, as in this embodiment, the size of the chip substrate 1 (= the size of the thin film member 2) is 25 mm × 75 mm.
Then, as described above, 1,000 to 80,000
The hole density is preferably such that the number of the opening portions 3 is provided (that is, the spot density). With such a spot density, the test can be performed efficiently.

Furthermore, the bonding state of the probe DNA and the sample DNA is usually after scanning with the laser, but is analyzed by performing the image processing, the opening section 3, from the viewpoint of such images processing It is preferable that they are arranged at a uniform pitch. The thin film member 2 is attached to the chip substrate 1 with an adhesive, for example, and a portion inside the concave portion 3a for accommodating the probe solution at the joint between the chip substrate 1 and the thin film member 2 joined by the adhesive. In this case, since the adhesive and the probe solution may come into contact with each other, a water-resistant adhesive is used as such an adhesive. Also,
When performing the hybridization, the temperature of the atmosphere of the DNA chip is raised to a predetermined temperature at which the DNA binding reaction is likely to occur. Therefore, an adhesive having a heat-resistant temperature of 60 to 80 ° C. or more is used. The adhesive is not particularly limited as long as it has such water resistance and heat resistance. For example, maleic anhydride is preferable.

Here, a method for manufacturing the present DNA chip will be described with reference to the process chart of FIG. First, the chip substrate 1 will be described. First, in step A10, glass or resin is formed into a predetermined shape (here, 25 mm × 75 m
m plate shape). In particular, the case where a resin material is used for the chip substrate 1 will be described. The resin material has a predetermined thickness (here, 0.7
(approximately 1.6 mm to 1.6 mm), and then cut into predetermined plane dimensions.

In step A20, a surface treatment such as a hydrophilic treatment (for example, a coating treatment for immobilizing DNA) is applied to the chip substrate 1 molded into a predetermined shape. Next, the thin film member 2 will be described. The thin film member 2 is usually manufactured in a state in which a plurality of the thin film members 2 are continuously formed in steps A30 to A50, and in step A60 immediately before attaching the thin film member 2 to the chip substrate 1, one chip is formed. It is cut into a size substantially the same as the substrate (here, about 25 mm × 75 mm). That is, first, in step A30, the resin is formed into a thin film with a predetermined thickness (here, 3 μm to 500 μm), and then, in step A40, an adhesive is applied to one surface, and then in step A50, the resin is opened in a predetermined arrangement and quantity. The hole 3 is penetrated by laser processing or punching.

At this point, as described above, the plurality of thin film members 2 are formed continuously, and here, in a step not shown, the long thin film member 2 in which the processes of steps A30 to A50 are completed. ', For example, as shown in FIG.
A role as shown in FIG. Then, in step A60, a thin film member of a predetermined length is sequentially sent out from the roll 2 ', and cut at a portion indicated by a two-dot chain line in the drawing to thereby obtain a predetermined size corresponding to one chip substrate 1. The thin film member 2 is obtained.

Then, in step A70, the chip substrate 1
The thin film member 2 cut to a predetermined size is attached to the side on which the surface treatment is performed, and further, in step A80,
A probe solution containing DNA whose sequence is known is dropped by a spotting head (not shown) into a concave portion 3 a formed by the opening 3 of the thin film member 2 and the surface of the chip substrate 1.
The manufacture of the A chip is completed.

The DNA chip according to the first embodiment of the present invention is configured as described above, and is formed by the following method (a chemical reaction test method according to the first embodiment of the present invention).
As shown in the flowchart of FIG.
Is analyzed. That is, first, in step B10, DNs containing different probe solutions are used.
A sample solution containing DNA to be tested is dropped into the concave portion 3a of the A chip.

Then, in step B20, the surrounding temperature of the DNA chip is adjusted to a predetermined temperature (around 60 to 80 ° C.) so that the DNA binding reaction easily occurs, and hybridization is performed. At this time, a surface member 4 such as glass or a film is disposed on the surface of the thin film member 2 as shown by a two-dot chain line in FIG. Thereby, the recess 3
In addition to preventing the water in the DNA solution in a from evaporating in a high-temperature atmosphere, the DNA solution to be inspected dropped on the DNA chip is completely fixed in the concave portion 3a over the entire surface of the DNA chip. Make sure that it comes in contact with the probe solution.

In this case, if an adhesive layer is further provided on the surface of the thin film member 2, the adhesion to the surface member 4 can be easily performed. Instead of attaching the surface member 4 to the thin film member 2 as described above, for example, after accommodating a DNA chip in a bag made of a film material, the bag is sealed and the inside is evacuated. Then, the DNA chip may be sealed with this bag-like material so that the sample in the concave portion 3a and the surroundings are shut off.

Since the unknown DNA to be inspected is fluorescently labeled in advance, after the surface member 4 on the upper surface of the thin film member 2 is removed in step B30 and the DNA chip is washed, the DNA to be inspected is Only the portion of the probe solution to be compared which is bound to the DNA is in a fluorescently labeled state. By measuring the amount of fluorescence of the sample in each recess 3a, the probe solution in any of the recesses 3a can
It is detected whether it has bound to A.

That is, the probe solution is fixed on the coating film 1 a formed on the surface of the chip substrate 1. When the fixed probe solution and the DNA to be inspected are bound, the DNA to be inspected is The applied fluorescent label is fixed to the coating film 1a. Therefore, when the DNA chip is washed, the DNA to be inspected that has not bound to the probe solution is washed away, but the DNA to be inspected that has bound to the probe solution is fixed to the coating film 1a together with the fluorescent label, and By measuring the amount of fluorescence of the sample in 3a, it is possible to detect which probe solution of the concave portion 3a has bound to the DNA to be inspected, and the base sequence of the DNA to be inspected is analyzed based on this.

Then, by analyzing the base sequence of such DNA, gene analysis and, consequently, diagnosis of genetic diseases and the like are performed. Therefore, in the present DNA chip and the chemical reaction test using the present DNA chip, the following advantages can be obtained. That is, since the probe solution is accommodated in the concave portion 3a, there is an advantage that the mixing of the adjacent probe solutions can be prevented, and the distance between the concave portions 3a can be narrowed to make the spot density sufficiently high.

Further, since the DNA chip has a simple structure in which the thin film member 2 provided with a large number of apertures 3 is attached to the chip substrate 1, the production can be simplified and the productivity can be improved. There are advantages. Further, since the thin film member 2 is formed of resin and has hydrophobicity, even when the supply position is slightly shifted when the water-soluble probe solution is supplied to the concave portion 3a by the spotting head, the hydrophobic thin film member 2 is formed. There is an advantage that the probe solution is supplied to the predetermined concave portion 3a as if the probe solution is repelled.

In the above-described embodiment, when manufacturing a DNA chip, as shown in FIG. 3, a roll is formed by winding a long thin film member 2 ′ in which a plurality of thin film members 2 are continuously formed. Then, a predetermined amount of the thin film member is sent out from the roll 2 ′ and cut into a size corresponding to one chip substrate 1. For example, as shown in FIG. After the thin film member 2 ″ formed on the substrate 5 is attached to the mount 5, a cut is made in only the thin film member 2 ″ to a size corresponding to one chip substrate 1 as shown by a two-dot chain line in the drawing. The thin film member 2 may be peeled off from the mount 5 and attached to the chip substrate 1.

In the above embodiment, an example using DNA as a biological sample has been described. However, the biological sample may be an oligonucleotide other than DNA, such as RNA (ribonucleotide) or PNA (peptide nucleotide). Alternatively, a peptide (including a protein) formed from a plurality of amino acids may be used. The use of a chip using RNA or PNA is for gene analysis and diagnosis of a genetic disease in the same manner as in the above-described embodiment, and the use of a chip using a peptide is a variety of methods using an antigen-antibody reaction or an enzyme reaction. Diagnosis of the disease can be considered. (B) Description of Second Embodiment Next, as a second embodiment of the present invention, an opening provided in a thin-film member is formed as a flow path. A chip for synthesis (microreactor chip), a chemical synthesis test (chemical reaction test method) using the chip for chemical synthesis, and a thin film member used for the chip for chemical synthesis. This chip for chemical synthesis is used for, for example, ordinary organic and inorganic synthesis reactions, biochemical reactions using oligonucleotides, peptides, and the like. Hereinafter, these will be described. 6 and 7 are views showing an example of the chemical synthesis chip of the present embodiment.

As shown in FIGS. 6A and 6B, the chip for chemical synthesis of the present invention (hereinafter also simply referred to as a chip) [FIG. 6 (B) is larger in the thickness direction than in the width direction. This is an enlarged view shown in the figure), and includes a chip substrate 11 and a thin film member 12 stacked and attached to the chip substrate 11. An opening 13 is formed in the thin film member 12, and the opening 13 cooperates with the surface of the chip substrate 11 to form a flow path 14 through which a sample for chemical synthesis flows. Has become.

As shown in FIG. 6A, for example, the flow path 14 includes a plurality of (here, three) branch paths 14a to 14c,
The branch passages 14a to 14c are configured to include a merging passage 14d formed as a group. With such a configuration, when different samples are injected from the sample injection sections 14A to 14C provided at the upstream ends of the respective branch paths 14a to 14c, these samples join at the junction channel 14d to perform the synthesis reaction. And the predetermined synthesis reactant is combined with the combined flow path 14d.
Can be collected from a sample collecting section 14D provided at the downstream end. The sample for chemical synthesis is appropriately selected depending on the purpose. In addition to the reagents used in ordinary organic synthesis reactions, the DN described in the first embodiment is used.
A biological sample (biological sample) such as an oligonucleotide such as A, RNA, or PNA or a peptide (including a protein) may be used.

Plane dimensions (width and length) of chip substrate 11
The thickness and the thickness are appropriately determined according to the chemical synthesis reaction. For example, if a relatively long time is required to obtain a predetermined synthesis reaction, the flow path length of the merging channel 14d in which the synthesis reaction is performed is set to be relatively large. It is also conceivable that the time taken for the sample to reach the sample collection section 14D at the downstream end of the merged channel 14d can be increased, thereby allowing the progress from the start of the synthesis reaction to the completion thereof to be merged. Observation along the road 14d is also possible.

Here, the chip substrate 11 is formed of a material that is easy to process, and has a surface on the side to which the thin film member 12 is attached (the side in contact with the sample) so as not to affect the synthesis reaction. A surface layer 11a made of a material that does not chemically react with the sample flowing through the flow path 14 (for example, a corrosion-resistant metal such as stainless steel, glass, Teflon (registered trademark), titanium, ceramics, or the like) is formed. Of course, if the chip substrate 11 itself is made of such a material that does not chemically react with the sample, it is not necessary to provide the surface layer 11a.

In addition, the material of the chip substrate 11 is as follows.
For example, if the sample is a DNA solution, D in the first embodiment
The above-described resin material exemplified as the material of the chip substrate 1 of the NA chip (see FIG. 1) can be used, and the molding method of the chip substrate 1 can be used as it is. The thin film member 12 is set to have the same plane dimensions as the chip substrate 11, and has a thickness T2 of 3 μm to 500 μm.
m. Further, the thin film member 12 is coated with an adhesive on one surface, and is adhered to the chip substrate 11 on this surface. As such an adhesive, one having resistance to a sample is appropriately selected. As the material of the thin film member 12, a material that does not chemically react with the sample similarly to the chip substrate 11 is appropriately selected.

The opening 13 is formed through the thin film member 12 by laser processing or punching. The width (flow path width) of the opening 13 (flow path 14) is appropriately set according to design conditions, and is usually 3 μm to 3000.
μm, preferably 10 μm to 100 μm
It is 0 μm, more preferably 10 to 500 μm, and by making the channel width appropriate, the flow of the sample is ensured and the synthesis reaction with a small amount of the sample becomes possible.

The flow path length and path of the opening 13 (flow path 14) are appropriately determined according to the synthesis reaction, and are limited to those shown in FIGS. 6 (A) and 6 (B). It is not something to be done. The manufacturing process of such a chip for chemical synthesis is as follows:
Although it is determined according to the material of the chip substrate 11 and the thin film member 12, for example, when the chip substrate 11 and the thin film member 12 are formed of resin similarly to the DNA chip of the above-described first embodiment, The process of the DNA chip shown in FIG. 2 is substantially the same as the process without the spotting in step A80.

However, by immobilizing a detection reagent (for example, probe DNA or the like) in the middle or at the end of the flow channel, the reaction between the immobilized sample and the sample flowing through the flow channel is generated. It is also possible to detect an object, and in this case, the spotting process of step A80 is performed as in the first embodiment. The chip for chemical synthesis as the second embodiment of the present invention is configured as described above, and is shown in the flowchart of FIG. 7 by the following method (chemical reaction test method as the second embodiment of the present invention). The reaction test is performed as follows. That is, first, predetermined samples different from each other are injected into the sample injection units 14A to 14C (step C10). Then, these samples are separated from the branch paths 14a to 14a.
The mixture flows from 14c to the joining channel 14d and is mixed to cause a synthesis reaction (Step C20). Then, the sample eventually flows into the sample collecting section 14D at the downstream end of the combined flow path 14d, and the synthesized reactant is collected from the sample collecting section 14D and analyzed (Step C30).

Therefore, in the chemical synthesis chip and the chemical reaction test using the chemical synthesis chip, the following advantages can be obtained. That is, since the present chemical synthesis chip has a simple configuration in which the thin film member 12 provided with the opening 13 is simply attached to the chip substrate 11, there is an advantage that manufacturing can be simplified and productivity can be improved. is there.

Further, when a groove is formed on a chip substrate by etching or the like as a flow path for flowing a sample as in a conventional case, there is a possibility that the groove shape may vary in edge shape at the bottom of the groove. On the other hand, in the present chemical synthesis chip, a flow path 14 is formed by the opening 13 and the surface of the chip substrate 11 by attaching the thin film member 12 having the opening 13 to the chip substrate 11. , Such opening 1
Since 3 is formed penetrating through the thin film member 12, there is an advantage that the processing is easier and the processing shape can be stabilized as compared with the groove processing. This also has the advantage of minimizing the effect of chip manufacturing errors on the synthesis test. (C) Others The microreactor of the present invention is not limited to the above embodiments, and can be variously modified without departing from the spirit of the invention.

For example, by providing a plurality of independent openings in the thin film member and laminating the thin film members on the chip substrate, a plurality of independent reaction chambers are formed between the openings and the surface of the chip substrate. You may do it.
In this case, by performing a plurality of chemical reactions simultaneously and in parallel in one microreactor, the whole can be processed and inspected under the same conditions, and the optimum conditions can be easily determined.

In the above-described embodiment, the adhesive is applied to the thin film member and the thin film member is attached to the chip substrate. However, any structure can be used as long as the thin film member can be stably laminated on the chip substrate. Instead of the adhesive, a double-sided adhesive tape may be interposed between the thin film portion and the chip substrate. Alternatively, if the thin film member and the chip substrate are made of the same material, they may be welded. The welding method may be heat fusion, hoop molding, or a solvent. When a solvent is used, since the solvent volatilizes relatively early, the solvent must be applied to the thin film member and / or the chip substrate immediately before welding the thin film member and the chip substrate.

[0062]

As described in detail above, according to the microreactor chip of the first aspect of the present invention, the hole is formed by a simple structure in which the thin film member provided with the hole is simply laminated on the chip substrate. Since the sample can be accommodated in cooperation with the unit and the chip substrate, there is an advantage that the manufacturing operation can be simplified and the productivity can be improved.

The opening is provided by penetrating the thin film member. By forming the opening by penetrating in this manner, the processing becomes stable and the manufacturing accuracy is improved. There is an advantage that can be. By setting the thickness of the thin film member, that is, the depth of the sample accommodating portion formed by the opening portion of the thin film member and the chip substrate in the range of 3 μm to 500 μm, the sample can be stably accommodated, and There is an advantage that the amount of the sample to be accommodated can be minimized (claim 2).

By forming the opening provided in the thin-film member with a plurality of independent fine holes, the sample is stably accommodated in the fine holes provided in the thin-film member. There is an advantage that the spot density can be improved by preventing mixing of the sample placed in the sample, and a chemical reaction test can be efficiently performed (claim 3).

The opening size of the micropore is 10 μm to 500 μm.
By setting the value within the range, a sufficient spot density is secured, and there is an advantage that the number of samples that can be handled by one microreactor chip can be increased and the chemical reaction test can be efficiently performed (claim 4). By forming the opening as a flow path for flowing the sample, there is an advantage that a predetermined chemical reaction can be performed only by flowing a small amount of the sample (claim 5).

By setting the width of the flow channel in the range of 10 μm to 1000 μm, there is an advantage that the flow of the sample is ensured and the synthesis reaction can be performed with a small amount of the sample (claim 6). Genetic analysis and disease diagnosis can be advantageously performed by using a sample as an oligonucleotide or a peptide (claim 7).

According to the chemical reaction test method of the present invention described in claim 8, the chemical reaction test is performed using the microreactor chip according to any one of claims 1 to 7, so that a small amount of sample can be efficiently used. There is an advantage that a test can be performed. According to the thin film member for a microreactor chip according to the present invention, the microreactor chip is constituted by forming an opening portion used for accommodating the sample, dividing the hole portion and laminating the chip portion on the chip substrate. There is an advantage that the chip manufacturing operation can be simplified and the productivity can be improved.

[Brief description of the drawings]

FIG. 1 is a diagram showing a configuration of a DNA chip as a first embodiment of the present invention, where (A) is a schematic plan view thereof,
(B) is a schematic diagram which expands and shows the XX cross section of (A).

FIG. 2 is a schematic manufacturing process diagram of a DNA chip as a first embodiment of the present invention.

FIG. 3 is a schematic perspective view showing a configuration of a roll of a thin film member on the DNA chip as the first embodiment of the present invention.

FIG. 4 is a flowchart for explaining a base sequence analysis method using a DNA chip according to the first embodiment of the present invention.

5A and 5B are diagrams for explaining a modification of the method for manufacturing a DNA chip as the first embodiment of the present invention, wherein FIG.
FIG. 2 is a schematic plan view of the thin film member, and FIG. 2B is a schematic side view of the thin film member.

FIG. 6 is a view showing a configuration of a chip for chemical synthesis as a second embodiment of the present invention, wherein (A) is a schematic plan view,
(B) is a schematic diagram which expands and shows the X1-X1 cross section of (A).

FIG. 7 is a flowchart for explaining a reaction test method using a chip for chemical synthesis according to a second embodiment of the present invention.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1, 11 Chip board 1a, 11a Coating film 2, 12 Thin film member 2 'Roll of thin film member 2 "Thin film member 3, 13 Opening 3a Depression 4 Film 5 Mount 14 Flow path 14A-14C Sample injection part 14D Sample collection part 14a to 14c Branch path 14d Merging path

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) G01N 33/566 G01N 35/02 F 35/02 37/00 102 37/00 102 C12N 15/00 F F term (Reference) 2G042 AA01 BD19 HA02 2G058 CC02 CC08 EA01 EA11 FB01 GA02 4B024 AA11 CA01 CA04 CA11 HA14 4B029 AA07 AA21 AA23 BB15 BB20 CC03 CC09 CC10 FA12 4B063 QA01 QQ42 QQ52 QR32 QR55 QS25 QS34

Claims (9)

[Claims] An opening for accommodating a sample in cooperation with the chip substrate in a state where the thin film member is laminated on the chip substrate, comprising a chip substrate and a thin film member laminated on the chip substrate. A microreactor chip, wherein a portion is provided on the thin film member. 2. The thin film member has a thickness of 3 μm to 500 μm.
The microreactor chip according to claim 1, wherein: 3. An opening provided in the thin film member,
The microreactor chip according to claim 1, wherein the microreactor chip is constituted by a plurality of independent micropores. 4. The micropore having an opening size of 10 μm to 500 μm.
3. The microreactor chip according to claim 2, wherein the diameter is in a range of μm. 5. An opening provided in the thin film member,
The microreactor chip according to claim 1, wherein the microreactor chip is formed as a flow path for flowing the sample. 6. The width dimension of said flow path is 10 μm to 1000 μm.
The microreactor chip according to claim 5, wherein m is in the range of m. 7. The microreactor chip according to claim 1, wherein the sample is an oligonucleotide or a peptide. 8. A chemical reaction test method comprising performing a chemical reaction test using the microreactor chip according to any one of claims 1 to 7. 9. A thin film member for a microreactor chip, which is provided with an opening used to accommodate a sample, divided and laminated on a chip substrate.