JP2009250684A - Microchip - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a microchip capable of properly controlling the injection position of the liquid reagent in a liquid reagent holding part. <P>SOLUTION: The microchip is equipped with a first substrate and the second substrate being the transparent substrate laminated on the first substrate and contains a fluid circuit including a cavity part constituted of the groove formed to the surface of the first substrate and the surface of the second substrate or the cavity part constituted of the surface of the first substrate and the groove formed to the surface of the second substrate. The fluid circuit includes the liquid reagent holding part for housing the liquid reagent and the liquid reagent holding part includes a liquid reagent discharge port for leading out the liquid reagent and a projection part is installed on the base of the groove of the first substrate which constitutes the base of the liquid reagent holding part or on the surface of the first substrate in the vicinity of the liquid reagent discharge port. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a microchip useful as a micro-TAS (Micro Total Analysis System) suitably used for biochemical tests such as DNA, proteins, cells, immunity and blood, chemical synthesis, and environmental analysis. Relates to a microchip with a built-in liquid reagent, in which a liquid reagent for mixing or reacting with a sample to be tested or analyzed is previously incorporated in the microchip.

  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 within a chip of several centimeters square and several millimeters in thickness. However, it has many advantages such as being able to perform high-throughput, high-throughput testing, and obtaining test results immediately at the site where the sample is collected, and is suitably used for biochemical tests such as blood tests.

  The microchip usually has a fluid circuit inside thereof, and the fluid circuit is used to measure the sample introduced into the fluid circuit, to mix the sample (for example, blood, etc.) and the reagent. Various fluid treatments are performed. Such fluid treatment can be performed by applying a centrifugal force in an appropriate direction to the microchip.

  Among the microchips described above, the liquid reagent built-in microchip is a microchip in which a liquid reagent for mixing or reacting with a sample or a specific component in the sample is held in advance in the fluid circuit. One or a plurality of liquid reagent holding units for holding the liquid reagent are provided (see, for example, Patent Document 1 for a microchip having a liquid reagent holding unit). The liquid reagent built-in microchip has one or more liquid reagent injection ports penetrating to the liquid reagent holding part for injecting the liquid reagent into the liquid reagent holding part on one surface thereof. The liquid reagent inlet is sealed by, for example, sticking a sealing label (seal) or the like on the surface of the microchip after the liquid reagent is injected.

  Here, if the liquid reagent is located in the vicinity of the discharge port of the liquid reagent holding unit, for example, the liquid reagent may leak from the discharge port into the fluid circuit due to a minute change in temperature and pressure. There is a risk of adversely affecting the mixing and reaction, and adversely affecting the test / analysis results of the mixture of the specimen and the liquid reagent.

Therefore, when the liquid reagent is injected from the liquid reagent injection port, it has been desired to guide the liquid reagent to a position as far as possible from the vicinity of the discharge port in the liquid reagent holding unit.
JP 2007-285792 A

  The present invention has been made to solve the above problems, and an object of the present invention is to provide a microchip capable of appropriately controlling the injection position of the liquid reagent in the liquid reagent holding section.

  The present invention includes a first substrate and a second substrate that is a transparent substrate laminated on the first substrate, and includes a groove formed on the surface of the first substrate and a surface of the second substrate. Or a microchip including a fluid circuit composed of a cavity formed of a first substrate surface and a groove formed on the second substrate surface, the fluid circuit containing a liquid reagent A liquid reagent holding portion for conducting the liquid reagent, the liquid reagent holding portion having a liquid reagent discharge port for leading out the liquid reagent, and a groove bottom surface of the first substrate constituting the bottom surface of the liquid reagent holding portion. The present invention relates to a microchip in which a protrusion is provided on the surface of one substrate and in the vicinity of a liquid reagent outlet.

  Further, in the microchip of the present invention, the protrusion portion is a second bottom surface constituting the ceiling surface of the liquid reagent holding portion from the groove bottom surface of the first substrate constituting the bottom surface of the liquid reagent holding portion or the first substrate surface. It is preferable to have a sloped surface that slopes in the direction of the liquid reagent discharge port toward the substrate surface or the groove bottom surface of the second substrate.

  In the microchip of the present invention, the second substrate further includes a liquid liquid reagent injection port for injecting a liquid reagent, and the gradient surface is disposed below the introduction port in the thickness direction of the microchip. It is preferable.

  In the microchip of the present invention, it is preferable that the inclined surface has a larger area than the liquid reagent inlet.

  The microchip in the present invention can appropriately control the injection position of the liquid reagent in the liquid reagent holding part.

  The microchip of the present invention is a chip capable of performing various chemical synthesis, inspection / analysis, and the like using a fluid circuit included in the microchip. In one preferred embodiment, the first substrate and the first substrate It consists of the 2nd board | substrate which is a transparent substrate laminated | stacked and bonded together. The first substrate and the second substrate having grooves on the substrate surface are bonded together so that the groove-forming surface of the first substrate faces the second substrate. Grooves are formed on the first substrate side surface of the second substrate. Therefore, the microchip composed of the two substrates has the grooves provided on the second substrate surface and the first substrate. And a fluid circuit comprising a cavity formed from the surface of the substrate facing the second substrate.

  In another preferred embodiment, the microchip of the present invention includes a third substrate, a first substrate having grooves provided on both surfaces of the substrate, and a laminated and bonded structure so as to narrow the first substrate. The second substrate and the third substrate, which are transparent substrates formed. Such a microchip composed of three substrates is composed of a groove provided on the surface of the second substrate facing the first substrate and the surface of the first substrate facing the second substrate. And a groove provided on the surface of the third substrate facing the first substrate and the surface of the first substrate facing the third substrate. And a second fluid circuit composed of a hollow portion and a two-layer fluid circuit. Here, “two layers” means that fluid circuits are provided at two different positions in the thickness direction of the microchip. The 1st fluid circuit and the 2nd fluid circuit may be connected by one or two or more penetration holes penetrated in the thickness direction formed in the 1st substrate.

  The method for bonding the substrates together is not particularly limited. For example, among the substrates to be bonded, at least one of the bonded surfaces of the substrates is melted and welded (welding method), and bonded using an adhesive. And the like. Examples of the welding method include a method in which the substrate is heated and welded; a method in which light is emitted from a laser or the like and the material is welded by heat generated during light absorption; and a method in which ultrasonic waves are used.

  The size of the microchip of the present invention is not particularly limited, and can be, for example, about several cm in length and width and about several mm to 1 cm in thickness.

  The material of each substrate constituting the microchip of the present invention is not particularly limited. For example, polyethylene terephthalate (PET), polybutylene terephthalate (PBT), polymethyl methacrylate (PMMA), polycarbonate (PC), polystyrene (PS) ), Polypropylene (PP), polyethylene (PE), polyethylene naphthalate (PEN), polyarylate resin (PAR), acrylonitrile-butadiene-styrene resin (ABS), vinyl chloride resin (PVC), polymethylpentene resin (PMP) Organic materials such as polybutadiene resin (PBD), biodegradable polymer (BP), cycloolefin polymer (COP), and polydimethylsiloxane (PDMS); inorganic materials such as silicon, glass, and quartz can be used.

  In the case where the microchip is composed of two sheets of the first and second substrates, the second substrate that is laminated on the first substrate and has a groove on the surface is preferably a transparent substrate. This point will be described in detail later with reference to an embodiment. The first substrate may be a transparent substrate, or may be a colored substrate such as a substrate made of a resin and a black substrate formed by adding carbon black or the like to the resin. It is preferable to use a black substrate. By using the first substrate as a colored substrate, a laser welding method using light such as a laser can be used. In addition, when bonding substrates by laser bonding, the bonded surface of the colored substrate is mainly melted and bonded, minimizing the deformation of the grooves formed on the transparent substrate. Can do.

  In the case where the microchip is composed of three sheets of the first substrate, the second substrate, and the third substrate, similarly, the second substrate stacked on the first substrate is a transparent substrate. The third substrate bonded to the lower side of the first substrate (the side opposite to the second substrate) is preferably a transparent substrate. As a result, as a part of the fluid circuit, a cuvette composed of a through hole penetrating the first substrate in the thickness direction and the transparent second and third substrate surfaces can be formed. Introduce a liquid mixture of the specimen and liquid reagent to be inspected and analyzed, and irradiate the cuvette with light in the direction perpendicular to the microchip surface from the upper surface (or lower surface) side of the microchip, and the opposite Optical measurement such as detecting the intensity (transmittance) of light transmitted from the liquid can be performed on the mixed solution. The first substrate positioned between the second substrate and the third substrate is preferably a colored substrate, and more preferably a black substrate.

  A method for forming grooves (flow path patterns) constituting a fluid circuit on the first substrate surface or the second substrate surface is not particularly limited, and an injection molding method using a mold having a transfer structure, an in- The printing method etc. can be mentioned. In the case of forming a substrate using an inorganic material, an etching method or the like can be used.

  In the microchip of the present invention, the fluid circuit (the first fluid circuit and the second fluid circuit in the case where two fluid circuits are provided) performs various processes appropriate for the liquid in the fluid circuit. In order to be able to do so, various parts arranged at appropriate positions in the fluid circuit are provided, and these parts are appropriately connected via fine flow paths.

  The microchip of the present invention is a microchip with a built-in liquid reagent in which a liquid reagent is previously held in the chip, and its fluid circuit is a liquid for holding the liquid reagent as one of the components constituting this. A reagent holding part is provided. There may be only one liquid reagent holding part, or two or more liquid reagent holding parts. A “liquid reagent” is a liquid substance for mixing or reacting with a specimen to be examined / analyzed. Only one type of liquid reagent may be incorporated in one microchip, or two or more types of liquid reagents may be incorporated. The “specimen” means a substance (for example, blood) to be tested and analyzed introduced into the fluid circuit itself or a specific component (for example, a plasma component) in the substance.

  The microchip of the present invention is usually a liquid reagent that is a through-hole penetrating to the upper surface (that is, the second substrate surface) up to the internal liquid reagent holding portion (penetrating the second substrate in the thickness direction). An inlet is provided. Such a microchip of the present invention usually has a label or seal for sealing the liquid reagent inlet on the microchip surface (second substrate surface) after the liquid reagent is injected from the liquid reagent inlet. Is affixed for use.

  In the present invention, the fluid circuit may include a part other than the liquid reagent holding part, such as a separation part for taking out a specific component from the specimen introduced into the fluid circuit; Sample measuring unit for measuring specific components. The same applies hereinafter.) Liquid reagent measuring unit for measuring liquid reagent; Mixing unit for mixing sample and liquid reagent; Examples include a detection unit (a cuvette for performing optical measurement) for performing inspection / analysis (for example, detection or quantification of a specific component in a liquid mixture). The microchip of the present invention may have all of these exemplified portions, or may not have any one or more. Moreover, you may have site | parts other than these illustrated site | parts. These portions are arranged at appropriate positions in the fluid circuit so as to perform a desired fluid treatment, and are connected through fine flow paths.

  The liquid mixture finally obtained by mixing the specimen and the liquid reagent is not particularly limited. For example, the liquid mixture that transmits the light by irradiating the portion (for example, the detection unit) containing the liquid mixture with light is transmitted. It is used for optical measurement such as a method for detecting intensity (transmittance), and inspection and analysis are performed.

  In a fluid circuit such as extraction of a specific component from a sample (separation of unnecessary components), measurement of a sample and / or a liquid reagent, mixing of a sample and a liquid reagent, introduction of the obtained mixture into a detection unit, etc. Various fluid treatments can be performed by sequentially applying a centrifugal force in an appropriate direction to the microchip. The application of the centrifugal force to the microchip can be performed by placing the microchip on a device (centrifuge) that can apply the centrifugal force. The centrifuge has a rotatable stage on a rotor (rotor). By placing a microchip on the stage and rotating the stage, the angle of the microchip relative to the rotor is arbitrarily certified. A centrifugal force can be applied in any direction.

  Here, the microchip of the present invention is provided with a protrusion in order to appropriately control the liquid reagent injection position in the liquid reagent holding part. Hereinafter, an embodiment will be shown and a microchip provided with a projection of the present invention will be described in detail.

<Embodiment>
FIG. 1 is a schematic top view showing an enlarged liquid reagent holding part in a microchip according to an embodiment of the present invention. FIG. 2 is a cross-sectional view taken along the line II-II in FIG. FIG. 3 is a schematic cross-sectional view showing the operation of the fluid in the microchip of the present embodiment. Hereinafter, description will be given with reference to FIGS. Further, in the present embodiment, “up in the thickness direction” means a direction on the second substrate 102 side in the thickness direction, and “down in the thickness direction” means on the first substrate 101 side in the thickness direction. It shall be the direction.

  First, the structure will be described with reference to FIGS. The microchip 100 of this embodiment is formed by bonding a second substrate 102, which is a transparent substrate, on a first substrate 101 having a groove on the surface, and the surface of the second substrate 102 on the first substrate 101 side. A fluid circuit including a groove formed on the surface of the first substrate 101 is included. FIG. 1 is an enlarged view showing the periphery of the liquid reagent holding unit 106 in the fluid circuit. Both the first substrate 101 and the second substrate 102 are plastic substrates made of an organic material as described above. The liquid reagent holding unit 106 is provided with a liquid reagent discharge port 104, and the liquid reagent in the liquid reagent holding unit 106 can be discharged by application of centrifugal force. In addition, a liquid reagent injection port 105 penetrating the second substrate 102 in the thickness direction is provided above the liquid reagent holding unit 106, and the liquid reagent is injected through the liquid reagent injection port 105. Note that the microchip 100 of the present embodiment may be a microchip including three substrates in which a third substrate (not shown) is bonded to the lower surface of the first substrate 101.

  Here, in the microchip 100, the protrusion 103 is provided on the bottom surface of the groove of the first substrate 101 constituting the bottom surface of the liquid reagent holding unit 106 and in the vicinity of the liquid reagent discharge port 104. More specifically, the protrusion 103 has a triangular cross section as shown in FIG. The vicinity of the liquid reagent discharge unit 106 can be set as appropriate, and the distance and the like are not particularly limited as long as the operation described later is guided. However, the distance L3 between the end portion of the liquid reagent discharge port 104 and the end portion of the liquid reagent discharge port 104 of the protrusion 103 can be set to, for example, 0.5 to 3 mm.

  1 has a length L1 in a first direction parallel to the liquid reagent discharge port 104 and a length L2 in a second direction which is a direction perpendicular to the second direction is 1 to 3 mm. Preferably there is. However, the length can be appropriately set to guide an operation described later.

  In the present embodiment, the protrusion 103 is formed on the surface of the second substrate 102 that forms the ceiling surface of the liquid reagent holding unit 106 from the groove bottom surface of the first substrate 101 that forms the bottom surface of the liquid reagent holding unit 106. Toward the liquid reagent discharge port 104.

  In the present embodiment, “gradient in the direction of the second substrate” means, for example, that the angle θ1 is less than 90 degrees. In the present embodiment, the angle θ1 is preferably 30 to 60 degrees. Furthermore, the angle θ1 is more preferably about 45 degrees. The range of the angle θ1 can be appropriately derived from the operation described later. The shape of the gradient surface itself is not particularly limited, and for example, it may be spherical.

  Here, the inclined surface is preferably inclined in the direction of the liquid reagent discharge port 104. Inclination in the direction of the liquid reagent discharge port 104 means, for example, the side far from the liquid reagent discharge port 104 in the cross section of the protrusion 103 in the first direction parallel to the liquid reagent discharge port 104 on the inclined surface (FIG. 2 (left side in 2) is at the lowest position, and the side closer to the liquid reagent discharge port 104 is at the highest position.

  In this embodiment, the second substrate 106 is provided with a liquid reagent injection port 105 for injecting a liquid reagent, but the gradient surface of the projection 104 is an introduction port in the thickness direction of the second substrate 102. It is installed under. Here, the inclined surface has a larger area than the liquid reagent introduction port 105. By having such a configuration, as will be described later, the liquid reagent injected from the liquid reagent inlet 105 can smoothly move away from the liquid reagent outlet 104 in the liquid reagent holding unit 106. It is.

  Next, the operation of the fluid in the present embodiment will be described based on FIG. First, the liquid reagent 202 is injected from the liquid liquid reagent inlet 105 using the nozzle 107. At this time, the liquid reagent 202 passes through the slope surface of the protrusion 103 and is guided in the direction opposite to the liquid reagent discharge port 104, that is, the left side in FIG. The liquid reagent 202 is maintained at an appropriate position in the liquid reagent holding unit 106.

  Further, even when the liquid reagent 202 is moved to the vicinity of the liquid reagent discharge port 104 of the liquid reagent holding unit due to an impact, a minute temperature or pressure change when the microchip is carried, the protruding portion 103 does not have the liquid reagent 202. Can be prevented from moving to the liquid reagent discharge port 104. That is, the length L3 in FIG. 1 is small enough to prevent the liquid reagent 202 from moving to the liquid reagent discharge port 104, and a centrifugal force was applied when measuring the sample with the microchip. At this time, the liquid reagent 202 needs to have a size that can move to the liquid reagent discharge port 104. The lengths L1 and L2 preferably have such a size that the protrusion 103 can cover the area of the liquid reagent inlet 105 immediately below the liquid reagent inlet 105 in the thickness direction. This is because the injected liquid reagent 202 can be guided and moved to a desired position. By the operation as described above, the amount of liquid reagent in the liquid reagent holding unit can be appropriately held.

  The microchip of the present embodiment can be variously modified without departing from the gist of the present invention. For example, the following modifications (1) to (2) can be performed. 4 and 5 are schematic cross-sectional views showing modifications of the microchip of the present embodiment.

  (1) Like the microchip 400 shown in FIG. 4, the grooves constituting the fluid circuit including the liquid reagent holding unit 406 may be formed not on the first substrate 401 but on the second substrate surface 402. . In FIG. 4, the fluid circuit including the liquid reagent holding unit 406 is formed by a groove formed on the surface of the second substrate 402 and the surface of the first substrate 401 substrate. The other configuration is the same as that of the microchip 100 shown in FIG. 1, and the liquid reagent holding unit 406 is provided with a projection 403 on the surface of the first substrate 401 and in the vicinity of the liquid reagent discharge port. . Then, the protrusion 403 extends from the surface of the first substrate 401 constituting the bottom surface of the liquid reagent holding unit 406 toward the groove bottom surface of the second substrate 402 constituting the ceiling surface of the liquid reagent holding unit 406. It has an inclined surface that is inclined in the direction of the discharge port 404. The second substrate 402 is formed with a liquid reagent inlet 405 that is a through hole penetrating in the thickness direction.

  (2) As in the microchip 600 shown in FIG. 5, the length in the thickness direction of the liquid reagent holding portion 606 on the first substrate 601 may be different from the length in the thickness direction at the liquid reagent discharge port 604. By taking such a structure, the liquid reagent can be further prevented from leaking to the liquid reagent discharge port 604 side. Since other structures (projections, etc.) are the same, description will not be repeated.

  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 a schematic top view which expands and shows the liquid reagent holding | maintenance part in the microchip of one Embodiment of this invention. It is sectional drawing along the II-II line in FIG. It is typical sectional drawing which shows the operation | movement of the fluid in the microchip of one Embodiment of this invention. It is typical sectional drawing which shows the modification of the microchip of one Embodiment of this invention. It is typical sectional drawing which shows the modification of the microchip of one Embodiment of this invention.

Explanation of symbols

  100, 400, 600 Microchip, 101, 401, 601 First substrate, 102, 402, 602 Second substrate, 103, 403 Protrusion, 104, 404, 604 Liquid reagent outlet, 105, 405, 605 Liquid Reagent introduction port, 106, 406, 606 Liquid reagent holding part, 107 nozzle, 202 Liquid reagent.

Claims (4)

A first substrate, and a second substrate that is a transparent substrate laminated on the first substrate,
It is comprised from the cavity formed from the groove | channel formed in the said 1st substrate surface, and the said 2nd substrate surface, or the groove | channel formed in the said 1st substrate surface and the said 2nd substrate surface. A microchip including a fluid circuit composed of a hollow portion,
The fluid circuit has a liquid reagent holding part for containing a liquid reagent,
The liquid reagent holding part has a liquid reagent outlet for leading out the liquid reagent,
A microchip in which a protrusion is provided on the bottom surface of the groove of the first substrate constituting the bottom surface of the liquid reagent holding unit or on the surface of the first substrate and in the vicinity of the liquid reagent discharge port.   The projecting portion extends from the groove bottom surface of the first substrate constituting the bottom surface of the liquid reagent holding portion or the surface of the first substrate, the second substrate surface constituting the ceiling surface of the liquid reagent holding portion, or 2. The microchip according to claim 1, wherein the microchip has a sloped surface that slopes in the direction of the liquid reagent outlet toward the bottom surface of the groove of the second substrate. A liquid reagent injection port for injecting a liquid reagent into the second substrate;
The microchip according to claim 1, wherein the inclined surface is installed below the introduction port in a thickness direction of the microchip.   The microchip according to claim 3, wherein the inclined surface has a larger area than the liquid reagent introduction port.

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