JP5215712B2 - Microchip - Google Patents

Description

  The present invention relates to a microchip.

  A microchip (microchannel chip) is known in which a flow path or the like through which a sample solution flows is formed on a substrate made of glass, silicon, resin, or the like using a micro-machining technology (for example, a patent) Reference 1).

  In a conventional microchip, a liquid feed port and a discharge port are provided in the microchip, and a sample solution is fed from a liquid feed port side by a drive source such as a pump. Alternatively, the sample solution is fed using the capillary force of the channel formed in the microchip.

(1) Disadvantages of pumping a sample solution with a pump or other drive source When using a pump, a drive source such as a pump is required. For each microchip, pump connection, pump removal, and pump output It is necessary to perform operations such as adjustment of the operation, which takes time.

(2) Disadvantages when Sample Solution is Delivered Utilizing Capillary Force of Flow Channel It is difficult to design a flow path, and in particular, it is difficult to manufacture a microchip having a complicated flow path pattern.

JP 2004-74339 A

  An object of the present invention is to provide a microchip that can be used easily and can be easily designed (manufactured) in various patterns.

Such an object is achieved by the present inventions (1) to (7) below.
(1) one or more sample supply ports;
One or more reaction vessels coated with reagents ;
One or more liquid supply paths connecting the sample supply port and the reaction tank or the reaction tanks;
One or more closure parts filled in the liquid feeding path,
The space including the reaction vessel formed by the closed portion is closed and depressurized,
When the closed part is damaged by irradiating light to the closed part and at least a part of the liquid feeding path penetrates, a sample is introduced into the reaction vessel with a pressure difference inside and outside the space. To microchip.

(2) one or more sample supply ports;
One or more reaction vessels coated with reagents ;
One or more suction tanks provided downstream of the reaction tank corresponding to the reaction tank;
Two or more liquid feed paths connecting the sample supply port and the reaction tank, the reaction tank and the suction tank;
One or more closed portions filled in a liquid feeding path connecting the reaction tank and the corresponding suction tank among the liquid feeding paths;
The space formed by the closing part and including the suction tank is closed and decompressed,
When the closed part is damaged by irradiating light to the closed part and at least a part of the liquid feeding path penetrates, a sample is placed in the reaction tank located upstream of the closed part due to a pressure difference inside and outside the space. Is introduced into the microchip.

  (3) The microchip according to (1) or (2), wherein the closing portion includes a light-absorbing substance.

(4) One of the microchip according to any one of the closure, characterized in that the Azukasuru the formation between a plurality of pre Kisora (1) to (3).

  (5) The microchip according to any one of (1) to (4), wherein the liquid supply path is branched and connected to the plurality of sample supply ports or the plurality of reaction vessels.

  (6) The microchip according to any one of (1) to (5) above, wherein a capturing unit that captures a specific component in the sample is provided in the liquid feeding path.

  (7) The microchip according to any one of (1) to (6), wherein the sample supply port is positioned above the reaction tank.

  According to the present invention, since the sample is transferred by the pressure difference, a driving source such as a pump is unnecessary. For this reason, operation is easy when using. In particular, a large number (large amount) of microchips can be processed (used) easily and quickly.

  In addition, the channel design is easier than when the sample is transferred using the capillary force of the channel. For this reason, for example, microchips having various patterns such as a microchip having a complicated flow path pattern can be easily manufactured.

  Moreover, since the space including the reaction tank formed by the closed part and the space including the suction tank formed by the closed part are closed and depressurized, it is difficult for changes with time to occur.

Embodiments of the present invention will be described with reference to the drawings.
First, the structure of the microchip according to the embodiment of the present invention will be described. FIG. 1 is a structural diagram of a microchip according to an embodiment of the present invention. FIG. 1A is a diagram (top view) of the microchip as viewed from above, and FIG. It is a cross-sectional view along arrow line AA ′ shown in FIG. FIG. 2 is an explanatory diagram of a sample inhalation method in the microchip of FIG.

  The microchip 1 mainly includes a substrate 2 and a cover plate 3. In the first microchip 1, a liquid feeding path 6 is formed in a groove shape on the substrate 2, and a hollow reaction tank 5 is formed by being connected to the liquid feeding path 6. The reaction tank 5 is coated with a reagent according to the purpose for detecting the reaction with the sample. In addition to the reagents, for example, nucleic acids, antigens, antibodies, and substances (for example, beads) that capture specific components in the sample can be applied to the reaction tank 5.

  In addition, the liquid supply path 6 is provided with a closed portion 7 in a form that occupies a part of the liquid supply path 6 so as to block the inflow of the sample. An inlet (sample supply port) 4 for injecting a sample is formed in the lid plate 3 so as to penetrate in the thickness direction.

  The microchip 1 has a structure in which the substrate 2 and the cover plate 3 are adhered and bonded so that the injection port 4 of the cover plate 3 is connected to the liquid feeding path 6 of the substrate 2. As a result, the closing portion 7 is in close contact with the substrate 2 and prevents the sample injected from the injection port 4 from flowing into the reaction vessel 5.

  In addition, the space closed by the closing portion 7, that is, a part of the liquid supply path 6 and the reaction tank 5 are depressurized and kept at a pressure lower than the atmosphere. In particular, the space is preferably in a vacuum or a pressure state close thereto. The closing part 7 contains a light-absorbing substance, and when the laser beam irradiated from the outside is absorbed, the closing part 7 is melted and damaged.

  Next, the manufacturing process of the microchip 1 will be described. First, the reaction tank 5 and the liquid feeding path 6 are formed on the substrate 2, and a closed portion 7 is filled in a part of the liquid feeding path 6. The reaction tank 5 and the liquid supply path 6 may be formed by etching, for example. A reagent is applied to the reaction tank 5 in advance.

  Next, the lid plate 3 on which the injection port 4 is formed is adhered and adhered to the substrate 2 at a position where the injection port 4 and the liquid supply path 6 are connected. In order to create a vacuum in the space from the liquid supply path 6 closed by the closing portion 7 to the reaction tank 5, the bonding process between the substrate 2 and the cover plate 3 is a work space maintained in a vacuum state or an environment close thereto. For example, in a vacuum chamber. In the bonding step, various methods such as adhesion using an ultraviolet curable adhesive, thermocompression bonding, or electrostatic bonding can be used.

  In the microchip 1 inhalation method, a sample injected from the injection port 4 is introduced into the reaction vessel 5, and as shown in FIG. 2, a laser beam emitted from a laser irradiation device (not shown) is emitted by the lens 10. The light is condensed and irradiated through the lid plate 3 toward the closing portion 7. When the closed portion 7 absorbs the laser beam and melts, and the liquid supply path 6 that has been divided by the closed portion 7 partially penetrates, the atmosphere is drawn into the reaction tank 5 due to the pressure difference from the atmospheric pressure. The sample is also sucked into the reaction tank 5 through the liquid feeding path 6. The sample sucked together with the air causes a chemical reaction with the reagent applied in the reaction tank 5, and the result is used for the component analysis of the sample.

  The microchip of the present invention does not have a sample outlet after the reaction, and is assumed to be disposable after one sample analysis. As a use of the microchip of the present invention, a method in which a reagent that reacts with the substance is previously applied to the reaction tank 5 according to the substance to be analyzed is selected and used.

  According to the microchip 1 of FIG. 1, the sample is sucked using the pressure difference between the closed space in the decompressed chip and the atmosphere, so that a driving source for sample suction such as a pump becomes unnecessary. Sample analysis can be performed with a simple configuration and operation.

  In the microchip 1, the material used for the closing portion 7 is a resin that contains a light-absorbing and heat-generating substance, for example, a pigment-based absorbing dye (such as carbon black) or a pigment-based absorbing dye and a dye-based absorbing dye. Is mentioned. On the other hand, the cover plate 3 is made of a light-transmitting substance (various glasses, polystyrene, polypropylene, polyamide, etc.). On the other hand, the substrate 2 does not necessarily need to use a light-transmitting substance, and a variety of materials such as various resins such as polyimide and silicon can be used as usual. Further, the cover plate 3 may be made of a light-transmitting material only in the portion irradiated with the laser beam, and a conventional material may be used in the other portions.

  As the laser beam, for example, a YAG (Yttrium Aluminum Garnet) laser or a laser diode can be used.

  Further, in FIG. 2, the laser beam is irradiated from the lid plate 3 side toward the closing part 7, but the irradiation direction of the laser beam may be from another direction, for example, from the substrate 2 side. Irradiation from a direction perpendicular to In this case, it is desirable that at least the material through which the laser beam passes is made of a light-transmitting substance.

  Furthermore, in order to smoothly inject the sample, the liquid supply path 6 may be formed so as to match the shape of the injection port 4 at a position connected to the injection port 4 in the substrate 2.

  Further, in the microchip 1 according to the embodiment of the present invention, a liquid feeding path 6 may be filled with a means (capturing means) that can capture a specific substance in a sample instead of a reagent, for example, beads. The beads are particles used for detection of a specific component in a sample, and have a structure in which a substance that binds to the specific component can be applied to the periphery. When the closed portion 7 is melted after the sample is injected, the beads are combined with specific components in the sample that has flowed in and pass through the liquid feeding path 6. If a member for damming the beads is provided in the liquid supply path 6, since the beads combined with the specific component are collected by the above member, whether or not the specific substance is contained in the sample by a reaction process using a colored substance or not. Can be determined.

  Next, a modification of the microchip according to the embodiment of the present invention will be described. FIG. 3 is a top view of a first modification of the microchip according to the embodiment of the present invention. Hereinafter, the microchips of a plurality of types of modified examples to be described later will be described mainly with respect to differences from the microchip of FIG. 1, and the same or corresponding parts as those of the microchip 1 of FIG. Description is omitted.

  The microchip 1 shown in FIG. 3 can detect a reaction of a sample in two stages, and has a configuration in which a substance reaction process can be continuously performed on one sample.

  The microchip 1 of FIG. 3 is provided with two reaction vessels, the first reaction vessel 51 is connected to the inlet 4 by a first liquid supply path 61, and the second reaction vessel 52 is a second reaction vessel 52. It connects with the 1st reaction tank 51 by the liquid feeding path 62. FIG. A reagent for detecting a reaction with the sample is applied to each of the first reaction tank 51 and the second reaction tank 52.

  Further, the first closing part 71 is filled in the first liquid supply path 61 and the second closing part 72 is filled in the second liquid supply path 62 in a part of each liquid supply path. Is provided. And the space (a part of the 1st liquid feeding path 61, the 1st reaction tank 51, a part of the 2nd liquid feeding path 62) closed by closed part 71 and closed part 72, and the 2nd closure The spaces closed by the section 72 (a part of the second liquid feeding path 62, the second reaction tank 52) are each decompressed to a pressure lower than the atmosphere.

  In the inhalation method of the microchip 1 of FIG. 3, in order to first introduce the sample injected from the injection port 4 into the reaction vessel 5, the laser beam is condensed and irradiated to the first closing part 71 through the cover plate. To do. When the first closing part 71 absorbs and melts the laser beam and the first liquid feeding path 61 partially penetrates, the atmosphere is drawn into the space from the first closing part 71 to the first reaction tank 51. Accordingly, the sample is also sucked into the first reaction tank 51 through the first liquid feeding path 61. The sample introduced into the first reaction tank 51 causes a chemical reaction with the reagent applied in the first reaction tank 51.

  After detecting the reaction in the first reaction tank 51, the laser beam is condensed and irradiated toward the second closing part 72 through the lid plate in order to introduce the sample into the second reaction tank 52. When the second closing portion 72 is melted and the second liquid feeding path 62 partially penetrates, the sample introduced into the first reaction tank 51 passes through the second liquid feeding path 62 together with the atmosphere to the second. The chemical reaction is caused with the reagent applied in the second reaction tank 52.

  As a matter of course, the microchip 1 shown in FIG. 3 can have a configuration in which three or more reaction vessels are connected in series through a liquid feed path. Also in this case, a reagent according to the purpose of analysis is applied to each reaction tank, the liquid supply path to each reaction tank is filled with a closed portion, and the space closed by each closed portion has a lower pressure than the atmosphere. The pressure is reduced to At the time of sample analysis, an operation of irradiating and melting each closed portion with laser in order close to the inlet 4 and introducing the sample into each reaction tank and reacting with the reagent are repeated.

  FIG. 4 is a top view of a second modification of the microchip according to the embodiment of the present invention. The microchip 1 in FIG. 4 has a structure in which the liquid feeding path is branched into two branches and each branch is connected to a corresponding reaction tank. Specifically, the liquid supply path 6 is connected to the inlet 4 through the main stream 60 and further branched into the tributaries 62a and 62b and connected to the reaction vessels 51 and 52, respectively. Further, the closing part 7 is provided so as to be filled in a part of the main stream 60 of the liquid supply path 6, and is a space closed by the closing part 7 (a space from a part of the main stream 60 to the reaction tanks 51 and 52. ) Is depressurized to a lower pressure than the atmosphere.

  The microchip 1 of FIG. 4 is melted by irradiating the closed portion 7 with laser, and when the main flow 60 of the liquid feeding path 6 partially penetrates, the sample from the inlet 4 passes through the main flow 60 together with the atmosphere to the tributaries 62a and 62b. And flow into the reaction tanks 51 and 52, respectively, and react with the reagents applied to the reaction tanks 51 and 52, respectively.

  The microchip 1 shown in FIG. 4 is suitable for performing two types of component analysis on an injected sample at the same time. For example, it is conceivable to apply different types of reagents to the reaction vessels 51 and 52. Of course, three or more reaction vessels may be provided, and the liquid feeding path 6 may be branched according to the number of reaction vessels.

  The liquid feed path 6 of the microchip in FIG. 4 is for branching and flowing the sample injected from one injection port into the two reaction vessels, but at the site of sample analysis, it differs from the plurality of injection ports. An analysis is also performed in which a sample is injected and the reaction is observed by flowing it into one or a plurality of reaction vessels at a time. Even in this analysis, the present invention can be applied by using a liquid feeding path branched toward the inlet side so that it can be connected to a plurality of inlets, and further providing a closing part upstream from the branch point. it can.

  FIG. 5 is a top view of a third modification of the microchip according to the embodiment of the present invention. In addition to the reaction tank 5, the microchip 1 of FIG. 5 is provided with a suction tank 8 corresponding to the reaction tank 5, and the reaction tank 5 is connected to the inlet 4 by a first liquid feeding path 61. The suction tank 8 is connected to the reaction tank 5 by the second liquid feeding path 62.

  In the microchip 1 of FIG. 5, the closing portion 7 is provided in a form that fills a partial section of the second liquid feeding path 62, and the space (the second liquid feeding path 62 of the second liquid feeding path 62 is closed). In part, the suction tank 8) is depressurized to a pressure lower than the atmosphere. A reagent for detecting a reaction with the sample is applied to the reaction tank 5.

  In the microchip 1 of FIG. 5, when the second liquid feeding path 62 partially penetrates the closed portion 7 by laser irradiation, the sample from the inlet 4 is sucked together with the atmosphere, and the first liquid feeding path It passes through 61 and reaches the reaction tank 5 and reacts with the reagent applied to the reaction tank 5. Further, the reagent passes through the second liquid feeding path 62 and finally reaches the suction tank 8.

  The microchip 1 in FIG. 5 is provided closer to the inlet 4 than the space in which the reaction tank 5 is decompressed, and flows into the reaction tank 5 in the process of sucking the sample into the suction tank 8 and is applied to the reaction tank 5. Reaction with the selected reagent occurs. When the closed portion 7 is irradiated with laser, impurities are generated as the closed portion 7 is melted and may be mixed into the sample. However, in the microchip 1 of FIG. Flow into 8. For this reason, reaction detection can be realized with a sample that does not contain impurities in the reaction vessel 5, and highly accurate sample analysis can be performed.

  FIG. 6 is a structural diagram of a fourth modification of the microchip according to the embodiment of the present invention. The microchip 1 in FIG. 6 has a rectangular parallelepiped shape, and a sample inlet 4 is opened on the upper surface, and is connected to a reaction tank 5 formed inside the microchip 1 through a liquid feeding path 6. . The reaction tank 5 is positioned vertically below the inlet 4.

  Moreover, the closed part 7 is provided in the form filled with the one part area of the liquid feeding path 6, and the space (part of the liquid feeding path 6, reaction tank 5) closed by the closed part 7 is more than air | atmosphere. Depressurized to low pressure. A reagent for detecting a reaction with the sample is applied to the reaction tank 5.

  When the microchip 1 in FIG. 6 is melted by irradiating the closed portion 7 with laser, the sample from the inlet 4 passes through the liquid feeding path 6 and reaches the reaction tank 5 by the suction force accompanying the gravity and pressure difference. Then, it reacts with the reagent applied to the reaction vessel 5.

  The microchip in FIG. 6 has a configuration in which the inlet 4 is positioned above the reaction tank 5, and therefore, when the closing part 7 melts and the liquid feed path 6 partially penetrates, the suction force due to the pressure difference is reduced. In addition, gravity applied to the sample itself acts and the sample flows into the reaction vessel 5. Therefore, the sample can be introduced into the reaction vessel 5 more rapidly than the case of only the suction force due to the pressure difference, and the analysis process can be speeded up.

  The manufacturing process of the microchip 1 in FIG. 6 may be performed by sealing and bonding two plate-like members in the same manner as in the case of the microchip in FIGS. You may make it form the injection port 4, the reaction tank 5, and the liquid sending path 6 by cutting the inside of a rectangular-shaped single member. In the case of the latter manufacturing process, it is preferable that after the inlet 4, the reaction tank 5, and the liquid feeding path 6 are formed, the closing portion 7 is pushed in from the inlet 4 to be brought into close contact with the liquid feeding path 6. .

  In the microchip 1 of FIG. 6, since the laser irradiates the closing part 7 from the side surface of the chip, it is desirable that the material of at least the portion through which the laser beam passes is a light-transmitting substance.

  6 may be applied to the microchip 1 shown in FIG. In particular, when the configuration of the microchip of FIG. 5 is applied, a suction tank and a second liquid feeding path connected to these are provided below the reaction tank 5, and the closing portion 7 is provided in a partial space of the second liquid feeding path. It becomes the structure provided so that it might be filled. According to this configuration, the reaction with the sample is performed at high speed and with high accuracy without the impurities generated by the laser irradiation to the closed portion 7 being mixed into the reaction vessel 5.

  Although the microchip according to the embodiment of the present invention has been described above, the above-described embodiment is merely an example, and the scope of the present invention is not limited to this. For example, the configuration of each part can be replaced with an arbitrary configuration having the same function, and other arbitrary configurations may be added.

  In addition, the present invention may be a combination of any two or more configurations (features) of the embodiment and its modifications.

1 is a structural diagram of a microchip according to an embodiment of the present invention. It is explanatory drawing of the inhalation method of the sample in the microchip of FIG. It is a top view of the 1st modification of the microchip concerning an embodiment of the invention. It is a top view of the 2nd modification of the microchip which concerns on embodiment of this invention. It is a top view of the 3rd modification of the microchip concerning an embodiment of the invention. It is a structural diagram of the 4th modification of the microchip which concerns on embodiment of this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Microchip 2 Substrate 3 Cover plate 4 Inlet 5 Reaction tank 51 1st reaction tank 52 2nd reaction tank 6 Liquid feed path 60 Main stream 61 1st liquid feed path 62 2nd liquid feed path 62a, 62b 7 Closing part 71 First closing part 72 Second closing part 8 Suction tank 10 Lens

Claims (7)

One or more sample supply ports;
One or more reaction vessels coated with reagents ;
One or more liquid supply paths connecting the sample supply port and the reaction tank or the reaction tanks;
One or more closure parts filled in the liquid feeding path,
The space including the reaction vessel formed by the closed portion is closed and depressurized,
When the closed part is damaged by irradiating light to the closed part and at least a part of the liquid feeding path penetrates, a sample is introduced into the reaction vessel with a pressure difference inside and outside the space. To microchip. One or more sample supply ports;
One or more reaction vessels coated with reagents ;
One or more suction tanks provided downstream of the reaction tank corresponding to the reaction tank;
Two or more liquid feed paths connecting the sample supply port and the reaction tank, the reaction tank and the suction tank;
One or more closed portions filled in a liquid feeding path connecting the reaction tank and the corresponding suction tank among the liquid feeding paths;
The space formed by the closing part and including the suction tank is closed and decompressed,
When the closed part is damaged by irradiating light to the closed part and at least a part of the liquid feeding path penetrates, a sample is placed in the reaction tank located upstream of the closed part due to a pressure difference inside and outside the space. Is introduced into the microchip.   The microchip according to claim 1, wherein the closing part includes a light-absorbing substance. The microchip according to any one of claims 1 to 3 one of the closure, characterized in that the Azukasuru the formation between a plurality of pre Kisora.   The microchip according to any one of claims 1 to 4, wherein the liquid supply path is branched and connected to the plurality of sample supply ports or the plurality of reaction vessels.   The microchip according to any one of claims 1 to 5, wherein a capturing means for capturing a specific component in the sample is provided in the liquid feeding path.   The microchip according to any one of claims 1 to 6, wherein the sample supply port is located above the reaction tank.

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