JP2009121985A - Microchannel chip, and apparatus and method for processing microchannel chip using microchannel chip - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a microchannel chip, and an apparatus and method for processing a microchannel chip, wherein the reaction is treated in a properly sealed condition, the sealing condition is maintained after reaction, and is not unlocked carelessly, and a space around an opening is secured even in a series of steps from introduction of a solution to reaction treatment. <P>SOLUTION: This method comprises: mounting a sealing lid 25 on a microchannel chip mounting stage 50 in an open state; introducing substances to be mixed by moving supply means 53, 54 of the substances closely to the opening in the open state; moving the sealing lid 25 to a sealing position of a fixing member 26 by a lid moving means 51 after the introduction of the substances is completed; ensuring the sealing of the fixing member 26 by performing the same operation on the sealing lid 25 in the sealed position as when the supply means 53, 54 of the substances have been moved closely to the opening; and processing the reaction and detection in a treatment position of a microchannel chip 100 by a processing means. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a microchannel chip used for analysis using various samples, a chemical reaction, and the like, a microchannel chip processing apparatus, and a microchannel chip processing method.

  In recent years, a method using a micro-channel chip has been proposed as a system that realizes analysis of a small amount of sample and chemical reaction processing inexpensively and quickly.

  Such a microchannel chip is applied to an inspection apparatus that supplies liquid to the chip and executes an inspection. As this inspection apparatus, for example, there is a biochemical processing apparatus disclosed in Patent Document 1, a stage on which a biochemical reaction cartridge (microchannel chip) having a chamber and a flow path communicating between the chambers is mounted, The moving means for moving the liquid through the path, the detecting means for detecting the presence / absence or amount of the liquid in the chamber, and the result of the liquid movement based on the information on the liquid in the chamber detected by the detecting means By providing the determination means, the sample preliminarily processed in the microchannel is guided into the chamber, and the sample is analyzed from the chemical reaction or biochemical reaction between the test reagent and the sample in the chamber. .

  Among such microchannel chips, in Patent Document 2, in a reaction vessel provided with a groove portion as an internal channel for solution, the reaction groove portion is designed for the purpose of suppressing evaporation loss of the solution introduced into the groove portion. A configuration in which the opening is provided with a lid has been proposed.

JP 2006-170654 A JP 2006-345813 A

By the way, when such a micro-channel chip is used for an inspection using an amplification reaction, it is possible that the user opens the lid after the reaction. In this case, the internal container may be blown out or spilled. There is a problem that contamination is high and contamination of the entire apparatus is caused.
In addition, the lids are individually closed, so that it is difficult to fix the position of the introduced liquid, and it cannot be used for reactions that require such solution position fixing.
In particular, in the case of Patent Document 2, since a space such as an opening connection port for introducing a solution is occupied by a lid, a series of actions as automation requiring a pump or the like cannot be taken.

  Therefore, the object of the present invention is to solve the above-mentioned problems, the reaction is processed in a reliably sealed state, and after the reaction, the sealed state is maintained, without being inadvertently opened, Also, by providing a microchannel chip that can secure a space around the opening in a series of steps from solution introduction to reaction processing, a microchannel chip processing apparatus, and a microchannel chip processing method. is there.

The above-described problems of the present invention are solved by the following configuration.
(1) A microchannel chip for mixing at least two substances to be mixed in a microchannel,
At least two openings provided on the same surface of the chip and communicating with the microchannel;
A microchannel chip comprising: a sealing lid having a fixing member that irreversibly seals after each mixed substance is introduced into each of the openings.

  According to the above configuration, the sealing lid having the fixing member that is irreversibly sealed after the introduction of the substance to be mixed is provided for each of at least two openings that are provided on the same surface of the microchannel chip and communicate with the microchannel. Therefore, the sealing lid does not open even during the reaction process, and since it is irreversibly sealed, the sealed state is maintained even after the reaction process, and it is opened carelessly even if the user is careless. Without contamination, it is possible to prevent device contamination and contamination.

(2) The microchannel chip according to (1), wherein the fixing member is a fixing hook provided in one sealing lid.
According to the above configuration, the fixing hooks that are the respective fixing members for the plurality of openings are provided on one sealing lid, so that at the time of sealing, the openings are simultaneously sealed, and the liquid is introduced into the microchannel. In this case, the simultaneous sealing hardly changes the internal pressure, and the liquid position at the time of sealing is maintained as it is. Therefore, this is particularly effective when the liquid position in the microchannel needs to be maintained at a fixed position.

(3) The microchannel chip according to (1) or (2), further including a sealing member that makes the opening watertight when the sealing lid is sealed.
According to the above configuration, the gap between the sealing lid and the opening can be completely closed by the sealing member that makes the opening watertight, and it is possible to more reliably maintain the internal pressure at the time of sealing, and the liquid leakage from the microchannel Can be eliminated without fail. Furthermore, in the case of heat treatment or the like, liquid leakage can be prevented to some extent even if the internal pressure increases.

(4) The microchannel chip according to any one of (1) to (3), wherein the sealing lid includes a hinge serving as a rotation shaft at an edge of the microchannel chip.
According to the above configuration, the rotation hinge of the sealing lid is the edge of the microchannel chip, so that the position when the sealing lid is opened is the lateral position of the microchannel chip and the surface on which the opening is arranged Can be placed at a distance from each other. Therefore, a sufficient space near the opening can be secured, and movement of equipment for introducing the substance to be mixed can be performed.

(5) The microchannel chip according to any one of the above (1) to (4), comprising a sheet that closes each opening and is peeled off when the substance to be mixed is introduced.
According to the above configuration, by providing a sheet that closes each opening and is peeled off when the substance to be mixed is introduced, contamination of the opening from the opening is prevented when handling the state before the reaction treatment, and further, the sealing lid is sealed before the treatment. Inadvertently sealing the opening with the fixing member can be avoided.

(6) A microchannel chip processing apparatus using the microchannel chip according to any one of (1) to (5) above,
A microchannel chip mounting table for mounting the sealing lid in an open state;
A mixed substance supply means capable of moving and closely contacting the opening position of the microchannel chip on the mounting table and capable of supplying a substance to be mixed from the opening;
Lid moving means that is provided in the mounting table and moves the sealing lid to the sealing position of the fixing member;
Processing means for performing a reaction and detection process on a processing position of the microchannel chip;
A microchannel chip processing apparatus comprising:

  According to the above configuration, the microfluidic chip mounted on the microfluidic chip mounting table is provided with the mixed substance supply unit, the lid moving unit, and the processing unit as described above, so that The reaction processing by the road chip can be performed efficiently and accurately while suppressing the risk of contamination and device contamination.

(7) The microchannel according to (6), wherein the mixed substance supply means includes a supply port connected to the opening and a vertical movement holding block that holds the supply port and is separated from and connected to the opening position of the microchannel chip. Chip.
According to the above configuration, the mixed substance supply means has the supply port connected to the opening and the vertical movement holding block, so that each movement of the mixed substance supply means when the microchannel chip is mounted and when the mixed substance is supplied This makes it possible to automate the supply of mixed substances.

(8) The microchannel chip according to the above (6) or (7), wherein the lid moving means is a rotary lever provided with a protrusion that contacts the lid.
According to the above configuration, after mounting the microchannel chip on the mounting table, the sealing lid can be moved to the sealing position by the rotation lever provided with the protrusion that contacts the lid, and in cooperation with the above (7), It becomes possible to automate the mixed substance supply and the opening sealing.

(9) A microchannel chip processing method using the microchannel chip or the microchannel chip processing apparatus according to any one of (1) to (8),
Place the sealing lid on the microchannel chip mounting table in an open state,
The mixed substance supply means is moved and brought into close contact with the open opening to introduce the mixed substance,
After the introduction of the mixed material, the sealing lid is moved to the sealing position of the fixing member by the lid moving means,
Secure the sealing of the fixing member by the same operation as the movement and adhesion of the mixed substance supply means on the sealing lid at the sealing position,
A microchannel chip processing method in which a reaction and detection process is performed on a processing position of a microchannel chip by the processing means.

  According to the above configuration, after placing the microchannel chip on the mounting table, introducing the substance to be mixed by the mixed substance supply means, and moving the sealing lid to the sealing position of the fixing member, in particular, the sealing position The sealing lid can be reliably sealed by pushing the sealing lid into the fixing member by the same operation as the movement / contact of the mixed substance supply means. Thereby, the process of the reaction and detection of the microchannel chip by the processing means can be performed efficiently and accurately while suppressing the risk of contamination and device contamination.

(10) When the microchannel chip according to (5) is mounted on the microchannel chip mounting table in a state where the sealing lid is opened, the sheet covering each opening is peeled off ( 9) The microchannel chip according to the above.

  According to the microchannel chip according to the present invention, the sealing lid does not open even during the reaction process, and since it is irreversibly sealed, the sealed state is maintained even after the reaction process, It is possible to prevent device contamination and contamination without being inadvertently opened even by the carelessness of the user. Further, the micro-channel chip processing apparatus and the micro-channel chip processing method can efficiently and accurately execute the reaction process and the sealing process while preventing the apparatus contamination and contamination.

Hereinafter, preferred embodiments of a microchannel chip, a microchannel chip processing apparatus, and a microchannel chip processing method according to the present invention will be described in detail with reference to the drawings.
FIG. 1 is an exploded perspective view of a microchannel chip according to the present invention, and FIG. 2 is a plan view illustrating the microchannel chip in a top view (a) and a bottom view (b).
As shown in FIG. 1, the microchannel chip 100 includes a channel substrate 21 and a lid member 23 attached to one surface (lower surface) 22 of the channel substrate 21. The flow path substrate 21 is manufactured by injection molding of a thermoplastic polymer. The polymer used is not particularly limited, but is preferably optically transparent, highly heat resistant, chemically stable, and easily injection-molded. COP, COC, PMMA, etc. are preferred. . Optically transparent means high transparency at the wavelength of excitation light or fluorescence used for detection, small scattering, and little autofluorescence. The chip 100 has translucency that enables detection of fluorescence. For example, cyber green is used as a detection reagent, and fluorescence emitted by intercalating into double-stranded DNA amplified by reaction is generated. It becomes possible to measure. Thereby, the presence or absence of the target gene sequence can be detected.

  The microchannel chip 100 is set and used on a mounting table (FIG. 3 to be described later) of the inspection apparatus, and is discarded for each use. In the present embodiment, blood (whole blood) as a specimen is injected into the microchannel chip 100. When the microchannel chip 100 is set in a testing device, a sample liquid is handled by a physical action force from the outside of the chip, for example, a single nucleotide polymorphism multiple target gene is tested. As shown in 2005-160387, a reaction for specifically amplifying a target sequence nucleic acid isothermally and its detection can be realized on the chip 1. This makes it possible to amplify and detect the target nucleic acid specific to the pathogen causing the infection, for example, by amplifying the target nucleic acid and detecting it, and determine the presence or absence of the pathogen in the sample. It becomes.

  In the present embodiment, the physical acting force is a pneumatic acting force (pneumatic driving force) generated by air supply or air suction from the port part PT provided at the start point and end point of the liquid flow path. Accordingly, the liquid supplied into the liquid flow path can be controlled to move to a desired position in the liquid flow path by air supply or air suction applied to the start point and end point of the liquid flow path. At this time, the liquid is held in a state of being sandwiched by the gas interposed between the start point and the front end portion and between the rear end portion and the end point, and is not divided in the middle by the action of the pulling force.

  The flow path substrate 21 includes ports, cells, flow paths, and the like for performing operations necessary for the liquid. That is, the flow path substrate 21 is provided with a first port PT-A for introducing a specimen liquid containing biological cells and a pretreatment reagent (first liquid), and a second port PT for introducing a reaction amplification reagent (second liquid). -D, the third port PT-B for supplying air pressure into the flow path, the fourth port PT-C at the end of the flow path to be depressurized, and the sample liquid introduced from the first port PT-A and pretreatment A first flow path (specimen mixing unit) A that mixes the reagent to generate a first mixed solution, and a second that extracts the DNA from living cells by heating the first mixed solution and decomposes it into single strands The flow path (heated part) B, the third flow path (reagent merging part) C for joining the reaction amplification reagent to the first mixed solution processed in the heated part B, and the reagent merging part C were joined. A fourth flow path (enzyme holding unit) D in which the enzyme (first solid) that is dissolved by passing the second mixed solution is solidified and mounted; and enzyme holding A fifth channel (enzyme mixing unit) E for promoting the mixing of the enzyme into the second mixed solution treated in D, and a primer (second unit) connected to the enzyme mixing unit E and solidified and mounted in the channel A plurality of sixth flow channels (reaction units) F that perform DNA amplification by heating, DNA amplification by heating, and detection of DNA amplification at the same position, and the enzyme mixing unit E connected to the flow channel of the reaction unit F A seventh flow path (quantitative dispensing flow path) G for quantitatively dispensing the second mixed liquid to each of the plurality of reaction detection cells 27 of the reaction section F is provided.

  Excavations 29 and 31 are formed in the other surface (upper surface) 28 of the flow path substrate 21, and the excavations 29 and 31 are located corresponding to the heated part B and the reaction part F. As shown in FIG. 2, the lower surface 22 of the flow path substrate 21 communicates with the first port PT-A, the second port PT-D, the third port PT-B, and the fourth port PT-C. Openings 33, 35, 37, and 39 are provided. The flow path substrate 21 has an outer shape of 55 × 91 mm in length and width W2, W1, and a thickness t of about 2 mm, for example.

Furthermore, a hinge 25a is provided on the side of the first port PT-A, the second port PT-D, the third port PT-B, and the fourth port PT-C at the edge of the upper surface 28 of the flow path substrate 21, and a sealing lid 25 is rotatably held. As the material, a sheet-like polymer that is optically transparent, has high heat resistance, and is chemically stable is used, like the channel substrate. The upper surface 28 side position of the sealing lid 25 is in a state of covering all of the first port PT-A, the second port PT-D, the third port PT-B, and the fourth port PT-C, and the first port PT-A, A seal member 25R that can seal each of the openings 33, 35, 37, and 39 in a watertight manner is disposed at a position facing each of the second port PT-D, the third port PT-B, and the fourth port PT-C. ing. Further, a plurality of fixing hooks 26 that are fixing members are arranged on the sealing lid 25, and hook fixing holes 27 are arranged at positions of the upper surface 28 that each of the fixing hooks 26 reaches.
As described above, since the fixing hook 26 and the hook fixing hole 27 are irreversibly sealed, the sealed state is maintained even after the reaction process, and the device is not opened carelessly even if the user is careless. Contamination and contamination can be prevented.

  Further, when the microchannel chip 100 is not used, a sheet 24 covering all of the first port PT-A, the second port PT-D, the third port PT-B, and the fourth port PT-C is attached and used. Sometimes taken away. With this sheet 24, it is possible to avoid fixing errors caused by the fixing hooks 26 when not in use.

  The lid member 23 is a member for covering ports, cells, and channels (grooves) formed on the channel surface (lower surface 22) of the channel substrate 21, and the lid member 23 and the channel substrate 21 are adhesives. And bonded with adhesive. As the lid member 23, a sheet-like high molecular polymer that is optically transparent, has high heat resistance, and is chemically stable is used, like the flow path substrate. In this embodiment, a PCR plate seal having a thickness of 100 μm is used (adhesive is applied to a plastic film). Furthermore, the width of the flow path is 1 mm, and there are some parts such as a mixing part that are thicker than that.

Next, the microchannel chip processing method of the present invention will be described below.
FIG. 3 is a schematic side view of the microchannel chip processing apparatus according to the present invention, and FIGS. 4A to 4F are side views showing processing steps in which only the microchannel chip is extracted.
The microchannel chip 21 with the sealing lid 25 opened is placed on the microchannel chip mounting table 50 of the microchannel chip processing apparatus 200. This position is determined in advance by a predetermined stopper or the like. A rotation lever 51 serving as a lid moving means is disposed on the mounting table 50, and the rotation shaft 51 a is pivotally supported by the mounting table 50 in the vicinity of the axis of the hinge 25 a of the microchannel chip 21. Then, the projection is not applied to the outer surface of the sealing lid 25 of the micro-channel chip 21, and the sealing lid is rotated by the rotation of the rotary lever 51 as indicated by an imaginary line (two-dot difference line). 25 is rotated to the sealing position of the microchannel chip 21.

  Above the micro-channel chip mounting table 50, a mixed substance supply unit including a supply port 54 and a vertical movement holding block 53 is supported by the frame 52. Further, a reaction detection sensor 55 is held by a holding frame 56 in the frame 52, and moves simultaneously with the vertical movement holding block 53 by the frame 52. The supply port 54 is provided corresponding to each of the first port PT-A, the second port PT-D, the third port PT-B, and the fourth port PT-C. The supply port 54 supplies a necessary material or gas to be mixed to each port.

  First, as shown in FIG. 4A, the microchannel chip 21 placed on the microchannel chip mounting table 50 has an open state in which the sealing lid 21 is disposed immediately beside the microchannel chip 21. It has become. At this time, the sheet 24 is stuck in a state where the ports PT-A, PT-D, PT-B, and PT-C are closed. Then, the sheet 24 is peeled off, and as shown in FIGS. 4B and 4C, the frame 52 of the mixed substance supply means is lowered, and the vertical movement holding block 53 and the supply port 54 correspond to the respective ports PT-A. , PT-D, PT-B, and PT-C, and supply the solution or sample L1. Next, in FIG. 4D, when the supply from the vertical movement holding block 53 and the supply port 54 is completed, the supplied internal liquid L2 reaches the processing position L3 through the flow path, and the frame 52 rises. The rotary lever 51 shown in FIG. 3 moves the sealing lid 25 by rotation, and rotates the sealing lid 25 to a position where the microchannel chip 21 is sealed. Then, the fixing hook 26 is inserted into the hook fixing hole 27.

  Here, when the fixing hook 26 is inserted into the hook fixing hole 27 by the rotating lever 51, the two members are weakly fixed and are not completely engaged. Therefore, as shown in FIG. 4E, the frame 52 is lowered again, and the sealing lid 25 is pushed down by the vertical movement holding block 53. By this operation, the engagement between the fixing hook 26 and the hook fixing hole 27 is completed. At this time, the reaction and detection at the processing position L3 are performed.

  FIG. 4F shows a state in which all the processes have been completed, and the engagement between the fixed hook 26 and the hook fixing hole 27 is a nonreciprocal engagement, and is discarded while this state is maintained.

  As described above, after the microchannel chip 100 is mounted on the mounting table 50, the substance to be mixed is introduced by the mixed substance supply means 53 and 54, and the sealing lid 25 is moved to the sealing position 27 of the fixing member 26. In particular, as shown in FIGS. 4D and 4E, the sealing lid 25 at the sealing position is pushed by the fixing member 26 by the same operation as the movement / adherence of the mixed substance supply means 53, 54, thereby sealing. The sealing of the lid can be ensured. Thereby, the process of the reaction and detection of the microchannel chip 100 by the processing means can be performed efficiently and accurately while suppressing the risk of contamination and device contamination.

It is a disassembled perspective view of the microchannel chip concerning the present invention. It is the top view which represented the top view of the microchannel chip | tip to (a), and the bottom view was represented to (b). It is a schematic side view of the microchannel chip processing apparatus according to the present invention. (A)-(f) is a side view which shows the process process which extracted only the microchannel chip | tip.

Explanation of symbols

21 flow path substrate 22 upper surface 23 lid material 24 sheet 25 sealing lid 26 fixing hook 27 hook fixing hole 28 lower surface 29, 31 digging 100 microchannel chip PT-A first port PT-B third port PT-C second 4 port PT-D 2nd port

Claims (10)

A microchannel chip for mixing at least two substances to be mixed in a microchannel,
At least two openings provided on the same surface of the chip and communicating with the microchannel;
A sealing lid having a fixing member for irreversibly sealing after introduction of the substance to be mixed for each of the openings;
A microchannel chip comprising:   The microchannel chip according to claim 1, wherein the fixing member is a fixing hook provided in one sealing lid.   The microchannel chip according to claim 1, further comprising a sealing member that seals the opening when the sealing lid is sealed.   The microchannel chip according to claim 1, wherein the sealing lid includes a hinge that serves as a rotation shaft at an edge of the microchannel chip.   The microchannel chip according to any one of claims 1 to 4, further comprising a sheet that closes the openings and is peeled off when the substance to be mixed is introduced. A microchannel chip processing apparatus using the microchannel chip according to any one of claims 1 to 5,
A microchannel chip mounting table for mounting the sealing lid in an open state;
A mixed substance supply means capable of moving and closely contacting the opening position of the microchannel chip on the mounting table and capable of supplying a substance to be mixed from the opening;
Lid moving means that is provided in the mounting table and moves the sealing lid to the sealing position of the fixing member;
Processing means for performing a reaction and detection process on a processing position of the microchannel chip;
A microchannel chip processing apparatus comprising:   The microchannel chip according to claim 6, wherein the mixed substance supply unit includes a supply port connected to the opening and a vertical movement holding block that holds the supply port and is separated from and contacted at the opening position of the microchannel chip. The microchannel chip according to claim 6 or 7, wherein the lid moving means is a rotary lever provided with a protrusion that contacts the lid. A microchannel chip processing method using the microchannel chip or the microchannel chip processing apparatus according to any one of claims 1 to 8,
Place the sealing lid on the microchannel chip mounting table in an open state,
The mixed substance supply means is moved and brought into close contact with the open opening to introduce the mixed substance,
After the introduction of the mixed material, the sealing lid is moved to the sealing position of the fixing member by the lid moving means,
Secure the sealing of the fixing member by the same operation as the movement and adhesion of the mixed substance supply means on the sealing lid at the sealing position,
A microchannel chip processing method in which a reaction and detection process is performed on a processing position of a microchannel chip by the processing means.   The microchannel chip according to claim 5 is peeled off from the sheet covering each opening when the sealing lid is placed on the microchannel chip mounting table in an open state. Channel chip.

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