JP2008134227A - Microchip and microchip inspection system - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a compact microchip capable of rapidly mixing a reagent when used with no scattering of a stored reagent. <P>SOLUTION: The microchip includes a reaction section in which at least one of a reagent and a sample from a passage is provided and a reaction thereof is performed by heating, in which a storage section for previously storing a reagent in the reaction section and a reagent previously stored in the storage section are sealed with a matter that changes a phase from a solid to a liquid between a storage temperature and a reaction temperature. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a microchip and a microchip inspection system thereof.

  In recent years, by making full use of micromachine technology and ultrafine processing technology, devices and means (for example, pumps, valves, flow paths, sensors, etc.) for performing conventional sample preparation, chemical analysis, chemical synthesis, etc. have been miniaturized. A system integrated on a chip attracts attention. This is also called μ-TAS (Micro Total Analysis System), and a reagent solution and a sample solution (for example, DNA extracted by processing a urine, saliva, blood, and sample of a subject to be examined) in a member called a microchip. This is a method for investigating the characteristics of a specimen by merging treated extraction solutions and the like and detecting the reaction.

  A microchip is based on a photolithographic process (a method of creating a groove by etching a pattern image with a chemical) on a substrate made of a resin material or a glass material, or by using a laser beam to form a groove. Various patterns have been proposed, in which a fine flow path through which a sample liquid and a reagent liquid can flow and a liquid reservoir for storing the reagent are provided.

  Furthermore, this μ-TAS is expected to be applied in the medical examination, diagnostic field, environmental measurement field, and agricultural production field. In reality, as is seen in genetic testing, when complicated processes, skilled techniques, and equipment operations are required, automation, acceleration, and simplification of the micro-analysis system make it costly and necessary. The analysis can be performed not only on the test amount and the time required, but also on any time and place, and the benefits are great.

In various types of analysis and inspection, importance is attached to the quantitativeness of analysis, the accuracy of analysis, and the economy of these microchips. For this purpose, a microchip having a simple configuration, high accuracy, and excellent reliability is required. The present inventors have already proposed a micropump system and a control method thereof suitable for this (Patent Documents 1 to 4).
JP 2004-28589 A JP 2001-322099 A JP 2004-108285 A JP 2004-270537 A

  In the analysis using the μ-TAS, it is desirable that a reagent is preliminarily sealed in a flow path formed on the microchip in order to perform analysis and inspection quickly. In this case, in an analysis that requires the use of a large number of reagents, it is necessary to provide a large number of flow paths for accommodating the reagents on the microchip. As a result, the microchip becomes large.

  In addition, when the reagent is sealed in advance in the microchip, the reagent is prevented from being scattered during storage before use, and the reagent is prevented from leaking from the storage part containing the reagent to the flow path communicating with the reagent. Is required. Furthermore, it is required that the reagent is quickly mixed at the time of use and that the reagent is smoothly flowed out from the storage unit containing the reagent to the subsequent flow path.

  An object of the present invention is to provide a microchip that is compact, does not scatter scattered reagents, and can quickly mix reagents when used.

The above object can be solved by the following configuration.
1. In a microchip having a reaction part in which at least one of a reagent and a sample is supplied from a flow path and a reaction is performed by heating,
The reaction unit has a storage unit for storing the reagent in advance,
The microchip, wherein the reagent stored in advance in the storage unit is sealed with a substance that undergoes a phase transition from solid to liquid between the storage temperature and the reaction temperature.
2. 2. The microchip according to 1, wherein the phase transition material is paraffin.
3. In a microchip having a reaction part in which at least one of a reagent and a sample is supplied from a flow path and a reaction is performed by heating,
The reaction unit has a storage unit for storing the reagent in advance,
The reagent stored in advance in the storage unit includes a substance that undergoes a phase transition from a solid to a liquid between a storage temperature and a reaction temperature.
4). 4. The microchip according to 3, wherein the phase transition material is gelatin.
5. The microchip according to any one of 1 to 4, wherein the storage unit is formed by providing a recess in a part of the reaction unit.
6.1 A microchip according to any one of 5 to 5;
A microchip inspecting device having a microchip accommodating portion that accommodates the microchip, and a heating unit that heats the reaction portion of the microchip when the microchip is accommodated in the microchip accommodating portion,
A microchip inspection system comprising:

  According to the present invention, since the reaction part is used as a reagent storage part, a compact microchip is obtained. In addition, during storage, the reagent in the reaction unit can be immobilized in the storage unit, and the reagent can be easily released from immobilization during use, so there is no scattering of the reagent during storage, and the reagent can be removed during use. Can be mixed quickly.

  An embodiment of the present invention will be described. In addition, although this invention is demonstrated based on embodiment of illustration, this invention is not restricted to this embodiment. In addition, the following assertive description in the embodiment of the present invention shows the best mode, and does not limit the meaning or technical scope of the terms of the present invention.

Hereinafter, embodiments of the present invention will be described with reference to the drawings.
(Device configuration)
FIG. 1 is an external view of an inspection apparatus 80 using the microchip according to the present embodiment. The inspection device 80 is a device that automatically reacts a sample and a reagent previously injected into the microchip 1 and automatically outputs a reaction result.

  The casing 82 of the inspection device 80 is provided with an insertion port 83 for inserting the microchip 1 into the device, a display unit 84, a memory card slot 85, a print output port 86, an operation panel 87, and an external input / output terminal 88. It has been.

  The person in charge of inspection inserts the microchip 1 in the direction of the arrow in FIG. 1 and operates the operation panel 87 to start the inspection. Inside the inspection device 80, the reaction in the microchip 1 is automatically inspected, and when the inspection is completed, the result is displayed on the display unit 84. The inspection result can be output from the print output port 86 or stored in a memory card inserted into the memory card slot 85 by operating the operation panel 87. Further, data can be stored in the personal computer or the like from the external input / output terminal 88 using, for example, a LAN cable. After completion of the inspection, the inspection person takes out the microchip 1 from the insertion port 83.

  FIG. 2 is a configuration diagram of an inspection apparatus 80 using the microchip according to the present embodiment. FIG. 2 shows a state where the microchip is inserted from the insertion port 83 shown in FIG. 1 and the setting is completed.

  The inspection apparatus 80 includes a driving liquid tank 10 that stores a driving liquid 11 for feeding a specimen and a reagent previously injected into the microchip 1, a micropump 5 for supplying the driving liquid 11 to the microchip 1, a micro pump A pump connection unit 6 that connects the pump 5 and the microchip 1 so that the driving liquid 11 does not leak, a temperature control unit 3 that controls the temperature of a necessary part of the microchip 1, and a temperature control unit 3 that prevents the microchip 1 from shifting. And a chip pressing plate 2 for closely contacting the pump connecting portion 6, a pressing plate driving unit 21 for raising and lowering the chip pressing plate 2, a regulating member 22 for accurately positioning the microchip 1 with respect to the micropump 5, and a microchip 1 includes a light detection unit 4 that detects a reaction state between the sample and the reagent in the sample 1.

  The chip pressing plate 2 is retracted upward from the position shown in FIG. 2 in the initial state. Thereby, the microchip 1 can be inserted / removed in the direction of the arrow X, and the person inspecting inserts the microchip 1 from the insertion port 83 (see FIG. 1) until it comes into contact with the regulating member 22. Thereafter, the chip pressing plate 2 is lowered by the pressing plate driving unit 21 and comes into contact with the microchip 1, and the lower surface of the microchip 1 is in close contact with the temperature adjustment unit 3 and the pump connection unit 6.

  The temperature control unit 3 includes a Peltier element 31 and a heater 32 on a surface facing the microchip 1. When the microchip 1 is set in the inspection device 80, the Peltier element 31 and the heater 32 are in close contact with the microchip 1. It is like that. The part containing the reagent is cooled by the Peltier element 31 so that the reagent is not denatured, or the part where the sample and the reagent react is heated by the heater 32 installed in the heating unit to promote the reaction. .

  The light detection unit includes a light emitting unit 4a and a light receiving unit 4b. The light detecting unit irradiates the microchip 1 with light from the light emitting unit 4a, and detects light transmitted through the microchip 1 with the light receiving unit 4b. The light receiving portion 4b is integrally provided inside the chip pressing plate 2. The light emitting part 4a and the light receiving part 4b are provided so as to face a detected part 148 (FIG. 3) of the microchip 1 described later.

  The micropump 5 is located on the pump chamber 52, the piezoelectric element 51 that changes the volume of the pump chamber 52, the first throttle channel 53 located on the microchip 1 side of the pump chamber 52, and the driving fluid tank 10 side of the pump chamber. The second throttle channel 54 is formed. The first throttle channel 53 and the second throttle channel 54 are narrow and narrow channels, and the first throttle channel 53 is longer than the second throttle channel 54.

  In the case where the driving liquid 11 is fed in the forward direction (direction toward the microchip 1), first, the piezoelectric element 51 is driven so as to rapidly reduce the volume of the pump chamber 52. Then, a turbulent flow is generated in the second throttle channel 54 that is a short throttle channel, and the channel resistance in the second throttle channel 54 is relatively larger than that of the first throttle channel 53 that is a throttle channel. growing. As a result, the driving liquid 11 in the pump chamber 52 is predominantly pushed toward the first throttle channel 53 and fed. Next, the piezoelectric element 51 is driven so that the volume of the pump chamber 52 is gradually increased. Then, the driving liquid 11 flows from the first throttle channel 53 and the second throttle channel 54 as the volume in the pump chamber 52 increases. At this time, since the length of the second throttle channel 54 is shorter than that of the first throttle channel 53, the channel resistance of the second throttle channel 54 is smaller than that of the first throttle channel 53. Thus, the driving liquid 11 flows predominantly from the second throttle channel 54 into the pump chamber 52. When the piezoelectric element 51 repeats the above operation, the driving liquid 11 is fed in the forward direction.

  On the other hand, when the driving liquid 11 is fed in the reverse direction (direction toward the driving liquid tank 10), first, the piezoelectric element 51 is driven so that the volume of the pump chamber 52 is gradually reduced. Then, since the length of the second throttle channel 54 is shorter than that of the first throttle channel 53, the channel resistance of the second throttle channel 54 is smaller than that of the first throttle channel 53. . As a result, the driving liquid 11 in the pump chamber 52 is predominantly pushed out toward the second throttle channel 54 and fed. Next, the piezoelectric element 51 is driven so that the volume of the pump chamber 52 is rapidly increased. Then, the driving liquid 11 flows from the first throttle channel 53 and the second throttle channel 54 as the volume in the pump chamber 52 increases. At this time, turbulent flow is generated in the second throttle channel 54, which is a short throttle channel, and the channel resistance in the second throttle channel 54 is relatively larger than that of the first throttle channel 53, which is a throttle channel. Become bigger. As a result, the driving liquid 11 flows into the pump chamber 52 predominantly from the first throttle channel 53. When the piezoelectric element 51 repeats the above operation, the driving liquid 11 is fed in the reverse direction.

The pump connection portion 6 is formed of a close contact surface made of a resin having flexibility (elasticity, shape followability) such as polytetrafluoroethylene or silicone resin in order to ensure necessary sealing performance and prevent leakage of driving fluid. It is preferred that Such a close contact surface having flexibility may be, for example, due to the constituent substrate of the microchip itself, or may be a separate additional having flexibility attached around the flow path opening in the pump connection portion 6. It may be due to a member.
(Configuration of microchip)
FIG. 3 shows an example of the configuration of the microchip 1 according to the present embodiment, and the present invention is not limited to this.

  The microchip 1 is provided with a flow path and a flow path element for mixing and reacting a liquid sample (specimen) on the microchip 1 as well as a liquid reagent. An example of processing performed in the microchip 1 by these flow paths and flow path elements will be described. The microchip 1 includes a groove forming substrate and a coated substrate that covers the groove forming substrate. FIG. 3 schematically shows the arrangement of the flow paths and flow path elements with the coated substrate removed. Yes. Further, in FIG. 3, an arrow indicates an insertion direction in which the microchip 1 is inserted into the inspection apparatus 80.

  Reference numeral 133 denotes a reagent storage unit that stores a reagent, and reference numeral 137 denotes a sample storage unit that stores a sample. In addition, openings 132 a and 132 b opened from one surface of the microchip 1 to the outside are provided upstream of the respective accommodating portions. These openings 132a and 132b are aligned with the flow path openings provided on the connection surface of the micropump 5 when the microchip 1 is connected to the micropump 5 through the pump connection portion 6 and connected to the micropump 5. The pump 5 communicates.

  A reaction unit 139 that mixes and reacts the reagent from the reagent storage unit 133 and the sample from the sample storage unit 137 is provided downstream of the reagent storage unit 133 and the sample storage unit 137.

  A detected part 148 is provided downstream of the reaction part 139, and a waste liquid part 60 is provided further downstream.

  The reagent stored in the reagent storage unit 133 flows into the reaction unit 139 by the driving liquid sent from the micropump 5 communicating with the opening 132a. On the other hand, the sample accommodated in the sample accommodating part 137 flows into the reaction part 139 by the driving liquid sent from the separate micro pump 5 communicating with the opening 132b. Thereby, in the reaction part 139, the reagent sent from the reagent storage part 133 and the sample sent from the sample storage part 137 are mixed.

The reagent and the sample mixed in the reaction unit 139 are heated by the heater 32 provided in the inspection apparatus 80 and the reaction is started. The liquid after the reaction is sent to the detected part 148. In the detected part 148, the target substance is detected by an optical detection method, for example. The liquid detected by the detected part 148 is sent to the waste liquid part 60.
(Configuration of the present invention)
Here, in the case where the reaction is performed by mixing with another reagent in addition to the reagent and the sample merged in the reaction unit 139, the flow path for injecting the reagent is insufficient. The first embodiment will be described with reference to FIG. FIG. 4 is a side sectional view of the reaction section 139. A recess is provided in a part of the reaction unit 139 to form a storage unit 150, and another reagent is stored in the recess. The reagent stored in the storage unit 150 is sealed with a sheet-like substance 151 that undergoes a phase transition from solid to liquid between the storage temperature and the reaction temperature.

  The phase transition sheet-like substance 151 is a paraffin having a melting point of about 20 to 60 ° C. and is an aliphatic hydrocarbon. Can be raised. Further, it may be a film of a compound that undergoes a sol-gel transition near 40 ° C., such as gelatin.

  A state in which a polymer exists in a colloidal form in a solution is called a sol. A gel indicates a state in which a polymer forms hydrogen bonds in an aqueous solution, and is formed by overcoming the Brownian motion of the polymer. As such a polymer exhibiting a sol-gel phase transition, gelatin and natural polysaccharides such as agarose are known. By cooling after dissolving in hot water, the liquid sol is changed into a soft solid gel. Phase transition.

  Since the sol-gel phase transition temperature varies depending on the type, when storing a reactive reagent, gelatin that undergoes sol-gel phase transition around 40 ° C. or low molecular weight agarose that undergoes sol-gel phase transition around 55 ° C. is preferably used as the reagent. be able to. When starting the reaction at a high temperature such as hot start PCR, high molecular weight agarose that undergoes a sol-gel phase transition around 80 ° C. can also be used. When the reagent is gelled, the reagent and sol can be mixed and gelled, but the sol is placed in the storage unit in advance, and after gelation, the reagent is added to diffuse the reagent into the gel. It is also possible to make it gel. In the latter case, the reagent is not brought to a high temperature state during the preparation of the reagent, which is a preferable form from the viewpoint of storage stability.

  Next, a second embodiment is shown in FIG. FIG. 5 is a side cross-sectional view of the reaction unit 139 showing a state in which gelatin dissolved at 50 ° C. is placed in a storage unit, gelled, a reagent is added, and the reagent is stored in the storage unit 150 in a gel state. FIG.

  In this case, after the liquid to be reacted with the gelled reagent is filled in the reaction part, the reaction part is heated to 40 ° C. or higher to make the gel into a sol, thereby mixing and reacting. For example, in the case of a PCR reaction, the temperature can be suddenly raised to 98 ° C. to start the reaction.

  Furthermore, in addition to the second embodiment, the third embodiment may be sealed with a sheet material 151 (not shown).

  As described above, the microchips of Embodiments 1 to 3 were manufactured, inserted into the inspection apparatus 80, and verified whether or not the reagent and the sample reacted in the reaction section at the heating temperature of 55 ° C. As a result, it was confirmed that there was no abnormality and it was functioning normally.

It is an external view of the test | inspection apparatus using the microchip which concerns on this embodiment. It is a block diagram of the test | inspection apparatus using the microchip which concerns on this embodiment. It is a block diagram of the microchip which concerns on this embodiment. It is side surface sectional drawing of the microchip which shows 1st Embodiment. It is side surface sectional drawing of the microchip which shows 2nd Embodiment.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Microchip 4 Photodetection part 5 Micropump 60 Waste liquid part 80 Inspection apparatus 132 Opening 133 Reagent storage part 137 Sample dissolution part 139 Reaction part 144 Reagent inlet part 145 Reagent storage part 147 Sample inlet part 148 Detected part 150 Storage part 151 Phase transition sheet material

Claims (6)

In a microchip having a reaction part in which at least one of a reagent and a sample is supplied from a flow path and a reaction is performed by heating,
The reaction unit has a storage unit for storing the reagent in advance,
The microchip, wherein the reagent stored in advance in the storage unit is sealed with a substance that undergoes a phase transition from solid to liquid between the storage temperature and the reaction temperature. The microchip according to claim 1, wherein the phase transition material is paraffin. In a microchip having a reaction part in which at least one of a reagent and a sample is supplied from a flow path and a reaction is performed by heating,
The reaction unit has a storage unit for storing the reagent in advance,
The reagent stored in advance in the storage unit includes a substance that undergoes a phase transition from a solid to a liquid between a storage temperature and a reaction temperature. The microchip according to claim 3, wherein the phase transition material is gelatin. The microchip according to claim 1, wherein the storage unit is formed by providing a recess in a part of the reaction unit. The microchip according to any one of claims 1 to 5,
A microchip inspecting device having a microchip accommodating portion that accommodates the microchip, and a heating unit that heats the reaction portion of the microchip when the microchip is accommodated in the microchip accommodating portion,
A microchip inspection system comprising:

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