JP5137010B2 - Microchip manufacturing method - Google Patents

Description

  The present invention relates to a microchip useful as a micro-TAS (Micro Total Analysis System) used for biochemical tests such as DNA, proteins, cells, immunity and blood, chemical synthesis, and environmental analysis, and a method for producing the same. .

  In recent years, the importance of detecting, detecting or quantifying biological substances such as DNA (Deoxyribo Nucleic Acid), enzymes, antigens, antibodies, proteins, viruses and cells, and chemical substances in fields such as medicine, health, food, and drug discovery There have been proposed various biochips and microchemical chips (hereinafter collectively referred to as microchips) that can be easily measured.

  Microchips can perform a series of experiments and analysis operations performed in the laboratory in a small-sized chip, so that only a small amount of sample and reagent are required, cost is low, reaction rate is high, and high-throughput inspection is performed. It has many advantages such as being able to obtain test results immediately at the site where the sample is collected, and is suitably used for biochemical tests such as blood tests.

Such a microchip includes a fluid circuit therein, and the fluid circuit is suitable for a specimen (substance to be examined / analyzed, such as blood) introduced into the fluid circuit. Various parts are provided so that processing can be performed, and these parts are appropriately connected by fine flow paths (see, for example, Patent Document 1 as an example of a microchip having a fluid circuit therein). .
JP 2006-239538 A

  The microchip including the fluid circuit includes, for example, a first substrate, a second substrate on which a pattern groove forming the fluid circuit is formed, and a third substrate, the first substrate and the third substrate. Thus, the second substrate can be sandwiched and bonded. At this time, the groove forming surface of the second substrate is previously provided with a surface such as a water repellent agent in order to improve the controllability of the liquid moving in the fluid circuit and prevent the liquid from remaining on the inner wall surface of the fluid circuit. A treatment agent may be applied.

  Here, as a method of bonding the three substrates, a welding method in which bonding is performed by melting the bonding contact surface of at least any one of the substrates can be suitably used. If the surface treatment agent is applied to the bonding contact surfaces of the substrate with the first and third substrates, it may be difficult to weld the substrates, and therefore the surface treatment agent layer on the bonding contact surface. Need to be removed. As a method for removing the surface treatment agent layer, for example, after applying the coating solution containing the surface treatment agent to both surfaces that are the groove forming surface of the second substrate, the coating solution layer is applied without drying. A method of wiping off the coating solution layer on the mating contact surface is mentioned. However, this method has a problem that even the coating liquid applied to the grooves constituting the fluid circuit is wiped off, or the coating liquid applied to the grooves adheres to the bonding contact surface. there were.

  Another method for removing the surface treatment agent layer on the bonding contact surface is to apply the coating solution containing the surface treatment agent to the entire groove-forming surfaces of the second substrate, and then to apply the coating solution layer. Examples thereof include a method of drying and polishing a coating film made of a dried surface treatment agent on a bonding contact surface. However, this method has a problem that particles generated by polishing may adhere to the fluid circuit.

  The present invention has been made to solve the above problems, and an object of the present invention is to provide a microchip in which the entire groove of the second substrate constituting the fluid circuit is reliably surface-treated. Another object of the present invention is to provide a method for manufacturing a microchip in which the entire groove of the second substrate is reliably surface-treated without generating particles and with sufficiently high bonding between the substrates. It is to be.

  The present invention is a microchip in which a first substrate, a second substrate having grooves provided on both sides of the substrate, and a third substrate are bonded together in this order, and the second substrate Provided is a microchip in which the side wall surface and the bottom surface of the groove have a coating film containing a surface treatment agent. Here, in this invention, it is preferable that the side wall surface and bottom face of all the grooves which the 2nd board | substrate has are coat | covered with the coating film containing a surface treating agent.

The second substrate has a through hole penetrating in the thickness direction, and the inner wall surface of the through hole is preferably covered with a coating film containing a surface treatment agent.
Moreover, it is preferable that the contact surface with a 1st board | substrate and the contact surface with a 3rd board | substrate in a 2nd board | substrate do not have a coating film (or surface treatment agent) containing a surface treatment agent.

  In the present invention, the second substrate can be a black substrate, and the first substrate and the third substrate can be transparent substrates. Moreover, as said surface treating agent, a water repellent treating agent can be mentioned suitably, for example. The thickness of the coating film is preferably 0.01 to 10 μm.

Further, according to the present invention, the first substrate, the second substrate having the grooves provided on both surfaces of the substrate, and the third substrate are bonded together in this order, and the grooves of the second substrate are provided. There is provided a method for manufacturing a microchip in which a side wall surface and a bottom surface are coated with a coating film containing a surface treatment agent, and the method includes the following steps.
(1) The process of apply | coating the coating liquid containing a surface treating agent on both surfaces of the said 2nd board | substrate,
(2) A step of drying the applied coating solution to obtain a coating film,
(3) A step of wetting the coating film on the surface that comes into contact with the first substrate surface and the third substrate surface during the bonding of the substrates, using a liquid, among the coating films,
(4) removing the wet coating film, and
(5) A step of bonding the first substrate, the second substrate, and the third substrate.

  The liquid is preferably a coating solution containing the surface treatment agent or a solvent capable of dissolving the surface treatment agent. As the surface treatment agent, for example, a water repellent treatment agent can be preferably exemplified.

  The thickness of the coating film obtained in the step of drying the coating solution to obtain a coating film is preferably 0.01 to 10 μm.

  The step of bonding the first substrate, the second substrate, and the third substrate preferably includes a step of melting the bonding surface of at least one of the substrates, and the second substrate is a black substrate. More preferably, at least the bonding surface of the second substrate is melted. The bonding surface of the substrates can be melted using, for example, a laser.

  ADVANTAGE OF THE INVENTION According to this invention, the microchip by which the groove | channel of the 2nd board | substrate which comprises a fluid circuit was reliably surface-treated can be provided. In addition, it is possible to provide a microchip with sufficiently high bondability between substrates without generating particles.

<Microchip>
The microchip of the present invention relates to a microchip formed by bonding a first substrate, a second substrate having grooves provided on both surfaces of the substrate, and a third substrate in this order. Such a microchip of the present invention includes a first fluid circuit including a second substrate side surface of the first substrate and a groove provided on the first substrate side surface of the second substrate, and a third fluid circuit. A two-layer fluid circuit including a second substrate side surface of the substrate and a second fluid circuit composed of grooves provided on the third substrate side surface of the second substrate is provided. Here, two layers means that fluid circuits are provided at two different positions in the thickness direction of the microchip. The first channel and the second channel may be connected to each other by one or two or more through holes penetrating in the thickness direction formed in the second substrate.

  In the microchip of the present invention, the fluid circuit (the first channel and the second channel) can perform various appropriate processes on the fluid (particularly, liquid) in the fluid circuit. Various parts are provided at appropriate positions in the fluid circuit, and these parts are appropriately connected through fine flow paths.

  The site is not particularly limited, but a liquid reagent holding unit for holding a liquid reagent, a measuring unit for measuring a sample or a liquid reagent, and for mixing a measured liquid reagent and a sample And a detection unit for performing inspection / analysis (for example, detection of a specific component in the mixed solution) of the mixed solution. Further parts may be provided as necessary. Here, in this specification, “specimen” refers to a sample itself (for example, blood) to be examined / analyzed introduced into a fluid circuit or a specific component (for example, blood sample) separated from the sample in the microchip. Plasma component separated from blood).

  The measuring unit has a predetermined capacity, and a predetermined amount of the sample or liquid reagent can be measured by introducing the sample or liquid reagent into the measuring unit. A liquid reagent is a reagent that processes a sample to be tested or analyzed using a microchip, or is mixed or reacted with the sample. Usually, a fluid circuit is prepared in advance before using the microchip. Built in the liquid reagent holding unit. The object to be measured such as the liquid mixture introduced into the detection unit is not particularly limited. For example, for optical measurement such as a method of detecting the intensity (transmittance) of light transmitted to the detection unit by irradiating light. The specimen is examined and analyzed.

  Fluid processing in the fluid circuit, such as weighing of specimens and liquid reagents, mixing of specimens and liquid reagents, and introduction of the obtained mixture into the detection unit, centrifugal force in the appropriate direction with respect to the microchip Can be applied sequentially. Application of centrifugal force to the microchip is typically performed by placing the microchip on a device (centrifuge) that can apply centrifugal force thereto.

  Hereinafter, the present invention will be described in detail with reference to embodiments. 1A and 1B are outline views showing an example of a microchip according to the present invention. FIG. 1A is a top view, FIG. 1B is a side view, and FIG. 1C is a bottom view. A microchip 100 shown in FIG. 1 is formed by laminating a first substrate 101 that is a transparent substrate, a second substrate 102 that is a black substrate, and a third substrate 103 that is a transparent substrate in this order (FIG. 1). (See (b)). The vertical and horizontal lengths of these substrates are not particularly limited, but in this embodiment, the horizontal (A in FIG. 1) is approximately 62 mm × vertical (B in FIG. 1) is approximately 30 mm. In this embodiment, the thicknesses of the first substrate 101, the second substrate 102, and the third substrate 103 (C, D, and E in FIG. 1 are about 1.6 mm, about 9 mm, and about 1. However, it is not limited to these.

  The first substrate 101 has liquid reagent inlets 110 (11 in total in the present embodiment) penetrating in the thickness direction and specimen inlets 120 for introducing specimens (for example, blood) into the microchip fluid circuit. Is formed. The microchip 100 is usually put into actual use after injecting a liquid reagent from the liquid reagent inlet 110 and then sealing the liquid reagent inlet 110 with a sealing label or the like.

  The second substrate 102 is formed with grooves formed on both surfaces thereof and a plurality of through holes penetrating in the thickness direction, and by bonding the first substrate 101 and the third substrate 103 thereto, The microchip 100 in which two layers of fluid circuits are formed is manufactured. Hereinafter, a fluid circuit composed of grooves provided on the surface of the first substrate 101 on the second substrate 102 side and the surface of the second substrate 102 on the first substrate 101 side is referred to as “first fluid circuit”, A fluid circuit including a groove provided on the second substrate 102 side surface of the third substrate 103 and a groove provided on the third substrate 103 side surface of the second substrate 102 is referred to as a “second fluid circuit”. These two fluid circuits are connected by a through-hole formed in the second substrate 102 that penetrates in the thickness direction. Hereinafter, the configuration of the two-layer fluid circuit included in the microchip 100 of the present embodiment will be described in detail.

  FIG. 2 is a perspective view showing a pattern of grooves formed in the second substrate 102. FIG. 2A is the first substrate 101 side (hereinafter, sometimes simply referred to as “upper side”). FIG. 2B is a perspective view showing a pattern of grooves formed on the surface, and FIG. 2B shows a pattern of grooves formed on the surface of the third substrate 103 side (hereinafter simply referred to as “lower side”). It is a perspective view. That is, FIG. 2A shows the first fluid circuit, and FIG. 2B shows the second fluid circuit. As shown in FIG. 2, the second substrate 102 is provided with each part where the specimen, the liquid reagent, or the mixed solution thereof is processed by the groove formed on the surface and the through hole penetrating in the thickness direction. A fine flow path that appropriately connects these portions is formed. The fluid circuit structure included in the microchip of the present invention is not limited to that shown in FIG. 2, and any fluid circuit structure may be used. The microchip 100 has a fluid circuit structure that can be suitably applied as a microchip that extracts a plasma component from blood and inspects and analyzes the plasma component.

  3 and 4 show a top view and a bottom view of the second substrate 102, respectively. FIG. 3 shows an upper fluid circuit (first fluid circuit) of the second substrate 102, and FIG. 4 shows a lower fluid circuit (second fluid circuit). In FIG. 4, the lower fluid circuit of the second substrate is shown in a horizontally inverted state so that the correspondence with the upper fluid circuit shown in FIG. 3 can be clearly understood. The microchip 100 of the present embodiment is a multi-item chip that can perform six items of inspection / analysis for one specimen, and the fluid circuit includes six items so that six items of inspection / analysis can be performed. It is divided into sections (sections 1 to 6 in FIG. 3) (however, they are connected to each other in the specimen measuring section installation area (lower fluid circuit upper area)). As described above, according to the present invention, since the fluid circuit has two layers, the fluid circuit can be integrated and densified. A microchip capable of inspection and analysis can be provided.

  In each of the above sections, one or two liquid reagent holding units each containing a liquid reagent are provided in the first fluid circuit (upper fluid circuit) (the liquid reagent holding units 301a and 301b in FIG. 3). , 302a, 302b, 303a, 303b, 304a, 304b, 305a, 305b and 306a in total 11). The sample introduced from the sample introduction port 120 in FIG. 1 is distributed to each section after the blood cell component is separated and removed, and when it is weighed, one or two kinds of liquid in each section weighed separately. It is mixed with the reagent and introduced into the detectors 311, 312, 313, 314, 315, and 316, respectively. The mixed liquid introduced into each detection unit of each section is used for optical measurement such as irradiating light to the detection unit from a direction substantially perpendicular to the microchip surface and measuring the transmittance of the transmitted light. The specific component in the mixed solution is detected. In these series of processes, by applying a centrifugal force in an appropriate direction to the microchip, the liquid reagent, the specimen, or a mixture thereof is supplied to each part in the two-layer fluid circuit provided in each section. This is done by moving them in an appropriate order. Application of the centrifugal force to the microchip can be performed, for example, by placing it on a centrifuge having a microchip mounting portion for placing the microchip. Each liquid reagent holding unit is connected to the liquid reagent measuring unit through a through hole penetrating the second substrate 102. In this way, by providing two layers of fluid circuits and connecting them by through holes, even a microchip having a relatively small area is moved between the first fluid circuit and the second fluid circuit. Thus, the fluid circuit can be used efficiently, and complicated liquid movement and the like can be controlled.

  Each section includes a total of the sample measuring units (sample measuring units 401, 402, 403, 404, 405, and 406 in FIG. 4) for measuring the sample in the second fluid circuit (lower fluid circuit). 6) and a liquid reagent metering unit for metering the liquid reagent (11 total liquid reagent metering units 411a, 411b, 412a, 412b, 413a, 413b, 414a, 414b, 415a, 415b and 416a in FIG. 4) are provided. ing. Each sample measuring section is connected in series by a flow path (see FIG. 4).

  Further, as shown in FIG. 3, the microchip 100 of the present embodiment overflows from the overflowing sample storage unit 330 for storing the sample overflowing from the sample measurement unit during measurement and from the liquid reagent measurement unit during measurement. There are overflow reagent storage portions 331a, 331b, 332a, 332b, 333a, 333b, 334a, 334b, 335a, 335b and 336a for storing the liquid reagent that has come out. The overflow specimen storage part 330 is connected to the specimen measurement part 406 via the flow path 16a (see FIG. 4), the through hole 26a penetrating in the thickness direction, and the flow path 16b (see FIG. 3). In addition, each overflow reagent storage unit is connected to a corresponding liquid reagent metering unit via a flow path and a through hole. For example, in the section 1, the liquid reagent measuring unit 411a for measuring the liquid reagent stored in the liquid reagent holding unit 301a and the overflow reagent storage unit 331a for storing the overflowing liquid reagent include the flow path 11a ( 4), and connected through a through hole 21a and a flow path 11b (see FIG. 3) penetrating in the thickness direction. The same applies to other overflow reagent storage units.

  Next, an example of fluid processing using the microchip 100 of the present embodiment will be described with reference to FIGS. 5 to 11 show the state of the liquid (specimen, liquid reagent and mixed liquid thereof) and the lower surface (third substrate side surface) of the upper surface (first substrate side surface) of the second substrate 102 in each step of fluid processing. It is a figure which shows the state of the liquid of (). (A) in each figure is a figure which shows the state of the liquid of the 2nd board | substrate upper surface (1st fluid circuit), (b) is the state of the liquid of the 2nd board | substrate lower surface (2nd fluid circuit). FIG. 5 (b) to 11 (b), as in FIG. 4, the left and right are reversed so that the correspondence with the upper fluid circuit shown in FIGS. 5 (a) to 11 (a) can be clearly understood. The lower fluid circuit of the 2nd board | substrate is shown in the state made to stand. Further, in the following description, only fluid processing in the fluid circuit of section 1 will be described, but the same processing is performed in other sections, and this can be clearly understood by referring to the drawings. Can do. Furthermore, in the following, a case where the specimen is blood (hereinafter, a plasma component separated from blood as defined above may also be referred to as a specimen) will be described as an example. It is not limited to this.

(A) Plasma separation and liquid reagent weighing step First, in this step, the microchip in the state shown in FIGS. 3 and 4 is directed downward in FIG. 5 (hereinafter simply referred to as downward. The same applies to FIGS. The same applies to the other directions.) A centrifugal force is applied. As a result, blood introduced from the sample introduction port of the first substrate 101 moves to the lower fluid circuit through the through hole 20a and is introduced into the plasma separation unit 420 (see FIG. 5B). The blood 600 introduced into the plasma separation unit 420 is centrifuged by the plasma separation unit 420 and separated into a plasma component (upper layer) and a blood cell component (lower layer). The blood overflowing from the plasma separation unit 420 moves to the upper fluid circuit through the through hole 20b and is stored in the waste liquid reservoir 430 (see FIG. 5A). Further, by applying a downward centrifugal force, the liquid reagents in the liquid reagent holding portions 301a and 301b reach the liquid reagent measuring portions 411a and 411b through the through holes 21b and 21c, respectively, and are weighed (FIG. 5B). reference). The liquid reagent overflowing from the measuring part passes through the through holes 21a and 21d, respectively, and is accommodated in the overflow reagent accommodating parts 331a and 331b in the upper surface side fluid circuit (see FIG. 5A).

(B) Specimen weighing process Next, a centrifugal force is applied to the left. As a result, the plasma component separated in the plasma separation unit 420 is introduced into the sample measurement unit 401 (at the same time, also introduced into the sample measurement units 402, 403, 404 and 405, 406) and is measured (FIG. 6 ( b)). The plasma component overflowing from the measuring portion is moved into the upper fluid circuit through the through hole 26a (see FIG. 6A).

(C) First mixing step Next, a downward centrifugal force is applied. As a result, the measured liquid reagent (the liquid reagent held in the liquid reagent holding unit 301a) and the plasma component measured by the sample measuring unit 401 are mixed in the liquid reagent measuring unit 411a (first). First step of mixing process, see FIG. 7B). At this time, a part of the liquid reagent remains in the mixing portion 441a of the lower fluid circuit. Next, by applying a leftward centrifugal force, the mixed solution is further mixed with the liquid reagent remaining in the mixing unit 441a (see the second step of the first mixing step, see FIG. 8B). These first step and second step are performed a plurality of times as necessary to ensure mixing. Finally, a state similar to the state shown in FIG. 8 is obtained.

(D) Second mixing step Next, an upward centrifugal force is applied. Thereby, the mixed liquid in the mixing unit 441a reaches the mixing unit 441b through the through hole 21e, and the other measured liquid reagent (the liquid reagent held in the liquid reagent holding unit 301b) is also It passes through the through hole 21e and reaches the mixing part 441b, and these are mixed (see the second step of the second mixing step, FIG. 9A). Next, by applying a rightward centrifugal force, as shown in FIG. 10A, the mixed solution moves in the mixing unit 441b and the mixing is promoted (second step of the second mixing step, FIG. 10 (a)). Further, the liquid reagent is accommodated in the overflow reagent accommodating portion 332b by the rightward centrifugal force (see FIG. 10A). These first step and second step are performed a plurality of times as necessary to ensure mixing. Finally, a state similar to the state shown in FIG. 10 is obtained.

(E) Detection part introduction process Finally, downward centrifugal force is applied. Thus, the mixed liquid is introduced into the detection unit 311 (refer to FIGS. 11A and 11B for other mixed liquids as well). The overflow reagent storage units 331a and 331b and the overflow sample storage unit 330 are in a state where a liquid reagent or a sample (plasma component) is stored. The same applies to other overflow reagent storage units. The mixed liquid filled in the detection unit is subjected to optical measurement, and a specimen (plasma component) is examined and analyzed. For example, a specific component in the mixed liquid can be detected by irradiating light from a direction substantially perpendicular to the surface of the microchip and measuring the transmitted light.

  Although the microchip of the present invention has been described with a preferred example, the fluid circuit structure included in the microchip of the present invention is not limited to the structure shown in the above embodiment. The microchip of the present invention is not necessarily a multi-item chip, and may be a single-item chip that performs only one type of inspection / analysis. Moreover, in this invention, it is not necessary to have all the above-mentioned each site | part, and it does not need to have any 1 or more types of site | parts. Or you may have the other site | part which is not mentioned above. Further, the number of each part included in the microchip is not particularly limited.

  Here, one feature of the microchip 100 of the present embodiment is that the side wall surface and the bottom surface of the groove of the second substrate 102 are covered with a coating film containing a surface treatment agent. FIG. 12 is a cross-sectional view schematically showing the microchip of this embodiment. However, the groove pattern formed on the second substrate 102 in FIG. 12 is merely schematically shown, and does not necessarily match the groove pattern shape shown in FIGS. As shown in FIG. 12, the side wall surface and the bottom surface of the groove on both surfaces of the second substrate 102 are covered with a coating film 1210 containing a surface treatment agent. In the present invention, the surface treating agent means an agent that imparts a specific property to the inner wall surface of the fluid circuit. Although it does not specifically limit as a surface treating agent, For example, protein adsorption inhibitors, such as water repellent treating agents, such as a fluorine-type resin and a silicone resin, MPC (2-methacryloyloxyethyl phosphorylcholine) polymer, etc. can be mentioned. By forming a coating film of water repellent treatment agent on the side wall and bottom surface of the groove on both sides of the second substrate, the controllability of the liquid (specimen, liquid reagent, and mixture thereof) moving in the fluid circuit is improved. In addition, it is possible to prevent the liquid from remaining on the inner wall surface of the fluid circuit. It is preferable that the coating film of the surface treatment agent is formed on the side wall surface and the bottom surface of all the grooves of the second substrate 102 so that the effect of the surface treatment agent is exhibited over the entire fluid circuit. Of the surface of the substrate (third substrate 103 in the present embodiment) located on the bottom surface when using the microchip, a coating film containing a surface treatment agent is also formed on the surface region constituting the inner wall of the fluid circuit. Also good.

  The thickness of the coating film 1210 is not particularly limited as long as a desired effect is obtained. For example, when forming a coating film of a water repellent treatment agent, the controllability of the liquid (sample, liquid reagent, and mixture thereof) moving in the fluid circuit is improved, and the liquid adheres to the inner wall surface of the fluid circuit. The thickness of the coating film is not particularly limited as long as an effect such as prevention of residual is obtained. The thickness of the coating film 1210 containing the surface treatment agent is, for example, 0.01 to 10 μm, and preferably 0.1 to 1 μm.

  The contact surfaces 1220a, 1220b, 1220c, 1221a, and 1221b of the second substrate 102 with the first substrate 101 and the third substrate 103 preferably do not have a coating film containing a surface treatment agent. Thereby, the bondability between the first substrate 101 and the third substrate 103 can be improved. In addition, when the substrates are bonded by melting the bonding surface of the second substrate 102, it is difficult to bond the substrates, or the bondability between the substrates tends to be low.

  The materials constituting the first substrate 101, the second substrate 102, and the third substrate 103 are not particularly limited, but considering the ease of forming the grooves constituting the fluid circuit, etc., the resin For example, polyethylene terephthalate (PET), polymethyl methacrylate (PMMA), polycarbonate (PC), polypropylene (PP), polystyrene (PS), polyvinyl chloride (PVC), cyclic polyolefin (COP), etc. Can be configured. The resin constituting the first substrate 101, the second substrate 102, and the third substrate 103 includes, in addition to the resin, a monomer component constituting the resin and another monomer component copolymerizable with the monomer component. A copolymer may also be used. In the case of using a colored substrate, a pigment component such as carbon black may be added to the resin. The materials of the first substrate 101, the second substrate 102, and the third substrate 103 may be different from each other or the same material.

  In the present invention, the first substrate and the third substrate do not necessarily need to be transparent substrates, but are transparent so that the transmitted light of the incident light can be measured at least in the region constituting the detection unit. In the case where the substrates are bonded by laser welding, which will be described later, it is more preferable to make both the substrates transparent. As a method for bonding the first substrate, the second substrate, and the third substrate, a bonding method (laser welding) such as bonding by irradiating and melting light (for example, laser light) on the bonding surface of the substrate Method), the second substrate is an opaque substrate (preferably a black substrate) and the first substrate and the third substrate are transparent substrates so that incident light can be absorbed more efficiently. Is preferred. Thereby, it is easy to bond the second substrate and the third substrate by irradiating light from the first substrate and the third substrate side and melting the bonding surface of the second substrate. Can be done.

<Microchip manufacturing method>
Next, the manufacturing method of the microchip of this invention is demonstrated in detail. The method for manufacturing a microchip according to the present invention includes a first substrate, a second substrate having grooves provided on both sides of the substrate, and a third substrate bonded together in this order. It can be suitably used for producing a microchip in which the side wall surface and the bottom surface of the groove of the substrate are covered with a coating film containing a surface treatment agent.

The manufacturing method of the microchip of the present invention includes the following steps.
(1) The process of apply | coating the coating liquid containing a surface treating agent on both surfaces of the said 2nd board | substrate,
(2) A step of drying the applied coating solution to obtain a coating film,
(3) A step of wetting the coating film on the surface that comes into contact with the first substrate surface and the third substrate surface during the bonding of the substrates, using a liquid, among the coating films,
(4) removing the wet coating film, and
(5) A step of bonding the first substrate, the second substrate, and the third substrate.
Hereinafter, each step will be described in detail with reference to FIG. FIG. 13 is a schematic process diagram showing an example of the manufacturing method of the microchip of the present invention, and shows schematic cross-sectional views of the substrate constituting the microchip at several stages during the manufacturing.

(1) Surface Treatment Agent Application Step In this step, first, a second substrate 1302 is prepared (FIG. 13A), and a coating solution containing a surface treatment agent is applied to both surfaces of the second substrate 1302. A second substrate 1302 shown in FIG. 13A is, for example, a black substrate, and has grooves 1304, 1305, and 1306 forming a fluid circuit on both surfaces thereof. The number and shape (width, depth, etc.) of the grooves are not particularly limited, and can be an appropriate pattern according to the desired structure of the fluid circuit. A substrate on which such a groove is formed can be produced using a mold having such a groove pattern transfer structure. Examples of the material of the second substrate 1302 include those described above.

  The coating liquid containing the surface treatment agent contains a solvent for dissolving, dispersing or diluting the surface treatment agent in addition to the surface treatment agent. Other additives may be added as necessary. The coating liquid used in the present invention includes a case where the surface treatment agent is dissolved in a solvent to form a solution, a case where the solid surface treatment agent is in the form of a slurry dispersed in the solvent, and the like. Specific examples of the surface treatment agent include those described above.

  The coating amount of the coating liquid is a surface treatment such as improving the controllability of the liquid (sample, liquid reagent, and a mixture thereof) moving in the fluid circuit and preventing the liquid from adhering to the inner wall surface of the fluid circuit. As long as the effect imparted by the agent is obtained, the coating amount is not particularly limited, and a coating amount that is usually employed in the field is employed. The coating amount of the coating solution is adjusted so that, for example, the thickness of the coating film after drying is about 0.01 to 10 μm, preferably about 0.1 to 1 μm.

  The method for applying the coating liquid is not particularly limited, and examples thereof include spray coating and dip coating. In spray coating, after the coating liquid is spray coated on one side of the second substrate, the substrate is turned over and spray coated again to obtain a second substrate having a coating liquid layer formed on both sides. . In addition, you may provide the process of wash | cleaning the surface where a coating liquid is apply | coated before the said application | coating process. As the cleaning liquid, for example, an alternative chlorofluorocarbon-based cleaning agent such as HFE (hydrofluoroether) can be used.

(2) Drying process Next, the applied coating liquid is dried to obtain a coating film 1310 containing a surface treatment agent (FIG. 13B). Drying can be performed under conditions of 20 to 80 ° C. using, for example, a blower. When the surface treatment agent removing step described later is performed without drying, not only the surface treatment agent on the contact surface with the first substrate and the third substrate but also the side wall surface and / or bottom surface of the groove is formed. The surface treatment agent may be removed, or the surface treatment agent formed on the side wall surface or the bottom surface of the groove may adhere to the contact surface between the first substrate and the third substrate. It is.

(3) Wetting step Next, among the coating films, contact surfaces with the first substrate and the third substrate (surfaces that contact the first and third substrate surfaces when the substrates are bonded) 1320a, 1320b, The coating films on 1320c, 1321a and 1321b are wetted with a liquid (FIG. 13 (c)). Although it does not restrict | limit especially as this liquid, The coating liquid itself containing a surface treating agent or the solvent contained in this coating liquid can be used suitably. It is also possible to use other solvents that can dissolve the coating film. By using such a liquid to wet the coating on the contact surfaces of the first substrate and the third substrate, the coating is typically dissolved by the liquid. Examples of the method of wetting the coating film with a liquid include a stamp method, a roller method, and a screen printing method.

By providing such a wetting step, the following problems can be avoided, and a flat bonded contact surface that does not have a coating film containing a surface treatment agent can be easily obtained.
(I) On the contact surface with the first and third substrate surfaces that may occur when the coating liquid is removed using a wiping paper or the like without drying the coating liquid containing the coated surface treating agent. In addition to the surface treatment agent, the surface treatment agent on the groove sidewall surface and / or bottom surface is removed, or the surface treatment agent on the groove sidewall surface or bottom surface is on the contact surface with the first and third substrate surfaces. The problem of sticking to, and
(Ii) Coating film particles, etc. that may occur when the coating liquid is removed by polishing the contact surfaces with the first and third substrate surfaces using polishing paper after drying the coating liquid The problem that occurs.

(4) Surface treatment agent removing step Next, the wetted coating film 1310a (see FIG. 13C) on the contact surface with the first and third substrate surfaces is removed (FIG. 13D). Thereby, the bondability between the second substrate, the first substrate, and the third substrate is improved. The wet paint film 1310a can be removed by, for example, a method such as “wiping” in which the second substrate 1302 is pressed and moved onto a wiping paper (wiper).

(5) Substrate Bonding Step Finally, the first substrate 1301 and the third substrate 1303 are bonded to the upper surface and the lower surface of the second substrate 1302, respectively (FIG. 13E). The first substrate 1301 and the third substrate 1303 do not necessarily have a flat plate shape, and grooves that can form a fluid circuit may be formed on the surfaces of these substrates. Examples of the material constituting the first substrate 1301 and the third substrate 1303 include those described above.

  The method for bonding the substrates is not particularly limited. For example, welding is performed by melting and bonding the bonding surfaces of at least one of the first substrate, the second substrate, and the third substrate. Method: Examples include a method of bonding using an adhesive. In consideration of suppression of flow path deformation at the time of bonding, protrusion of the adhesive into the fluid circuit, and adverse effects due to the reaction between the adhesive and the reagent, bonding is preferably performed by a welding method. Examples of the welding method include thermal welding, ultrasonic welding, and the like in addition to laser welding in which a bonded surface of a substrate is melted using a laser. Among these, it is preferable to use a laser welding method because only the vicinity of the bonding surface can be melted.

  In the laser welding method, typically, first, a first substrate 1301 and a second substrate 1302 that are transparent substrates are stacked while being aligned, and then laser light is emitted from the first substrate 1301 side. Is applied toward the substrate bonding surface, and the generated heat causes the bonding surface of one or both of the substrates at the interface between the substrates to melt. At this time, if the second substrate 1302 is a colored substrate, preferably a black substrate, the light absorption rate of the second substrate 1320 is higher than that of the first substrate 1301. The bonding surface of the substrate 1302 is melted. Next, the first substrate 1301 and the second substrate 1302 are bonded to the laminate of the first substrate 1301 by applying pressure from the upper and lower surfaces of the laminate through a glass substrate or the like. A bonded body with the second substrate 1302 is obtained. Next, after a third substrate 1303 is stacked on the second substrate 1302 of the bonded body, alignment is performed, and then laser light is irradiated from the third substrate 1303 side, whereby laser is similarly emitted. Welding is performed. Next, a microchip is obtained by applying pressure to the laminated body composed of the three substrates through a glass substrate or the like from the upper and lower surfaces of the laminated body.

  According to the above-described method of the present invention, the microchip having a high bondability between the substrates and having the surface treated reliably in the grooves of the second substrate can generate dust or dirt derived from the coating film. It becomes possible to manufacture without.

  The embodiment disclosed this time should be considered as illustrative in all points and not restrictive. The scope of the present invention is defined by the terms of the claims, rather than the description above, and is intended to include any modifications within the scope and meaning equivalent to the terms of the claims.

It is an outline view showing an example of a microchip concerning the present invention. It is a perspective view which shows an example of the pattern of the groove | channel formed in the 2nd board | substrate of the microchip based on this invention. It is a top view which shows an example of the 2nd board | substrate of the microchip based on this invention. It is a bottom view which shows an example of the 2nd board | substrate of the microchip which concerns on this invention. It is a figure which shows the state of the liquid of the upper surface (1st board | substrate side surface) of a 2nd board | substrate and the state of the liquid of a lower surface (3rd board | substrate side surface) in plasma separation and a liquid reagent measurement process. It is a figure which shows the state of the liquid of the upper surface (1st board | substrate side surface) of a 2nd board | substrate in the sample measurement process, and the state of the liquid of a lower surface (3rd board | substrate side surface). It is a figure which shows the state of the liquid of the upper surface (1st substrate side surface) of a 2nd board | substrate in the 1st step of a 1st mixing process, and the state of the liquid of a lower surface (3rd substrate side surface). It is a figure which shows the state of the liquid of the upper surface (1st substrate side surface) of a 2nd board | substrate in the 1st mixing process 2nd step, and the state of the liquid of a lower surface (3rd substrate side surface). It is a figure which shows the state of the liquid of the upper surface (1st substrate side surface) of a 2nd board | substrate in the 2nd mixing process 1st step, and the state of the liquid of a lower surface (3rd substrate side surface). It is a figure which shows the state of the liquid of the upper surface (1st substrate side surface) of a 2nd board | substrate in the 2nd step of a 2nd mixing process, and the state of the liquid of a lower surface (3rd substrate side surface). It is a figure which shows the state of the liquid of the upper surface (1st substrate side surface) of a 2nd board | substrate in the detection part introduction | transduction process, and the state of the liquid of a lower surface (3rd substrate side surface). It is sectional drawing which shows typically an example of the microchip which concerns on this invention. It is a schematic process drawing which shows an example of the manufacturing method of the microchip of this invention.

Explanation of symbols

  100 microchip, 101, 1301 first substrate, 102, 1302 second substrate, 103, 1303 third substrate, 110 liquid reagent inlet, 120 sample inlet, 130 recess, 301a, 301b, 302a, 302b, 303a, 303b, 304a, 304b, 305a, 305b, 306a Liquid reagent holding unit, 311, 312, 313, 314, 315, 316 detection unit, 330 overflow specimen storage unit, 331 a, 331 b, 332 a, 332 b, 333 a, 333 b, 334a, 334b, 335a, 335b, 336a Overflow reagent storage unit, 401, 402, 403, 404, 405, 406 Sample weighing unit, 411a, 411b, 412a, 412b, 413a, 413b, 414a, 414b, 415a, 415b, 416aBody reagent measurement unit, 420 Plasma separation unit, 430 Waste liquid reservoir, 441a, 441b Mixing unit, 11a, 11b, 16a, 16b Flow path, 20a, 20b, 21a, 21b, 21c, 21d, 21e, 26a Through hole, 510 Overflow Liquid container, 600 blood, 1210, 1310 Coating containing surface treatment agent, 1220a, 1220b, 1220c, 1221a, 1221b, 1320a, 1320b, 1320c, 1321a, 1321b Contact with first substrate and third substrate Surface, 1304, 1305, 1306 Groove, 1310a Wet coating.

Claims (7)

A first substrate, a second substrate provided with grooves provided on both sides of the substrate, and a third substrate are bonded together in this order, and the side wall surface and the bottom surface of the groove of the second substrate are A method for producing a microchip coated with a coating film containing a surface treatment agent,
Applying a coating solution containing a surface treatment agent on both surfaces of the second substrate;
A step of drying the applied coating solution to obtain a coating film;
Of the coating film, a step of wetting the coating film on the surface that comes into contact with the first substrate surface and the third substrate at the time of bonding of the substrate using a liquid;
Removing the wet coating;
Bonding the first substrate, the second substrate, and the third substrate;
A microchip manufacturing method including: The method for producing a microchip according to claim 1 , wherein the liquid is a coating solution containing the surface treatment agent or a solvent capable of dissolving the surface treatment agent. The surface treatment agent, a microchip manufacturing method according to claim 1 or 2 which is water repellent. The microchip manufacturing method according to any one of claims 1 to 3 , wherein a thickness of the coating film obtained in the step of drying the coating liquid to obtain a coating film is 0.01 to 10 µm. Step of bonding the first substrate and the second substrate and the third substrate, according to any one of claims 1 to 4, comprising the step of melting the bonding surface of at least one of the substrates Microchip manufacturing method. The method for manufacturing a microchip according to claim 5 , wherein the second substrate is a black substrate, and at least a bonding surface of the second substrate is melted. The method of manufacturing a microchip according to claim 5 or 6 , wherein the bonding surface of the substrate is melted by using a laser.

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