JP2009287971A - Microchip - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a microchip with a metering part which has a low metering error, satisfactory cutting-off between the liquid in a metering part body and the liquid discharged from the metering part body and is capable of suppressing an occupying area low. <P>SOLUTION: The microchip has a first fluid circuit containing a first substrate having a groove provided to its surface and/or having a through-hole piercing therethrough in its thickness direction and the second substrate laminated on the first substrate. The first fluid circuit includes the metering part for metering the liquid, and the metering part has the metering part body with an introducing port for introducing the liquid and a discharge port for discharging the liquid, the opening comprising the groove provided to the surface of the first substrate or the through-hole piercing the first substrate in the thickness direction thereof, and a connection flow channel for connecting the discharge port and the opening. The metering part body and the opening are arranged in opposed relationship so as to hold the connection flow channel therebetween. <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. Specifically, the present invention relates to a microchip including a measuring unit for measuring a liquid such as a specimen or a liquid reagent.

  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 cm square and a thickness of several millimeters to 1 cm. It has many advantages such as fast reaction speed, high-throughput testing, and the ability to obtain test results immediately at the site where the sample is collected. For example, it 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 or a specific component in the sample introduced into the fluid circuit, and the specific component in the sample or the sample. Various fluid treatments such as mixing with liquid reagents are performed. Such fluid treatment can be performed by applying a centrifugal force in an appropriate direction to the microchip.

When a sample to be examined or analyzed or a specific component in the sample and a liquid reagent are mixed in the microchip and the sample or the specific component is treated with the liquid reagent (or reacted with the liquid reagent), the micro The chip preferably includes a metering unit for metering the specimen or the specific component and / or a metering unit for metering the liquid reagent in order to mix them at an appropriate ratio. For example, Patent Document 1 (see in particular FIG. 3) includes a plasma measuring chamber for measuring plasma centrifuged from blood, and a buffer measuring chamber for measuring a buffer solution mixed with the plasma. A blanket is provided.
JP-A 64-25058

  An example of the structure of the measuring unit that measures the specimen (or a specific component in the specimen) and the liquid reagent that acts on the specimen (or a specific component in the specimen) is shown in FIG. The measuring unit 10 shown in FIG. 1 is connected to a measuring unit main body 20 that is a part (chamber) for measuring a liquid such as a specimen or a liquid reagent, and an opening 30 provided in the upper part of the measuring unit main body 20. A discharge channel 40 and a waste liquid reservoir 50 connected to the other end of the discharge channel 40 are provided. The waste liquid reservoir 50 stores the liquid with respect to the opening 30 so as to accommodate the excess liquid (waste liquid) that overflows when the liquid is introduced into the measurement body 20 and exceeds the capacity of the measurement body 20. It is arranged on the downstream side in the direction of the centrifugal force when introduced into 20. The liquid that has flowed from the direction of the arrow shown by applying centrifugal force to the microchip is introduced into the measuring unit main body 20 from the opening 30, and excessive liquid exceeding the capacity of the measuring unit main body 20. As waste liquid, it again flows through the discharge channel 40 through the opening 30 and is stored in the waste liquid reservoir 50. At this time, the measured liquid surface position in the measuring unit main body 20 is the illustrated position.

However, the measuring unit 10 shown in FIG. 1 has the following problems and has room for improvement.
(1) Since the opening 30 serves as both an inlet for introducing liquid and an outlet for discharging excess liquid, the opening 30 is used to satisfactorily introduce and discharge the liquid. Although it is necessary to make the width of 30 relatively wide, in this case, the surface area of the liquid surface of the weighed liquid becomes large, and the measurement error tends to increase. That is, when the surface area of the liquid surface of the measured liquid is large, a difference in the liquid amount based on the difference in the liquid surface shape appears remarkably, and the measurement error tends to increase.
(2) The liquid is introduced into the measuring unit main body 20 by application of the centrifugal force, and a part of the liquid is discharged from the opening 30 in the direction of the discharge flow path 40, but the measuring unit immediately after the application of the centrifugal force is stopped. There is a case where the liquid breakage between the liquid in the main body 20 and the liquid flowing out in the direction of the discharge flow path 40 is not good. May occur.
(3) Since the discharge flow path 40 is provided, the area occupied by the measuring portion in the fluid circuit becomes relatively large, which may be an obstacle to miniaturization of the microchip and the design of the fluid circuit structure.

  Further, a weighing unit having a structure as shown in FIG. 2 is also conceivable as a weighing unit that can solve the above problems. The metering unit 60 shown in FIG. 2 includes a metering unit main body 70 that is a part for metering liquid, and an outlet port 72, which includes an inlet 71 for introducing a liquid and a discharge port 72 for discharging the liquid. And a waste liquid reservoir 90 connected to the other end of the discharge flow path 80. The waste liquid reservoir 90 is similar to the waste liquid reservoir 50 when the liquid is introduced into the measuring unit main body 70 with respect to the discharge port 72 so that the waste liquid overflowed when the liquid is introduced into the measuring unit main body 70 can be accommodated. It is arrange | positioned in the downstream in the centrifugal force direction. The liquid that has flowed from the direction of the arrow shown by applying centrifugal force to the microchip is introduced into the measuring unit main body 70 from the introduction port 71, and an excess amount exceeding the capacity of the measuring unit main body 70. The liquid flows through the discharge channel 80 through the discharge port 72 and is stored in the waste liquid reservoir 90. At this time, the measured liquid surface position in the measuring unit main body 70 is the illustrated position.

  According to such a structure, an opening for introducing the liquid and an opening for discharging the liquid are separately provided, and the width of each opening is reduced, whereby the surface area of the liquid surface of the measured liquid can be reduced. Therefore, the weighing error can be further reduced. However, there is still a problem in terms of liquid breakage between the liquid in the measuring unit main body 70 immediately after the application of the centrifugal force and the liquid flowing in the direction of the discharge flow path 80 and the occupied area of the measuring unit. There is room for improvement.

  The present invention has been made in order to solve the above-mentioned problems, and its purpose is that the measurement error is small and the liquid breakage between the liquid in the measurement unit main body and the liquid discharged from the measurement unit main body is good. It is another object to provide a microchip including a measuring unit that can keep an occupied area low.

  The present invention includes a first substrate having a groove and / or a through-hole penetrating in the thickness direction on the surface, and a second substrate laminated on the first substrate, the groove possessed by the first substrate And a microchip having a first fluid circuit composed of a hollow portion constituted by a first substrate side surface of a second substrate. In the microchip of the present invention, the first fluid circuit includes a measuring unit for measuring the liquid, and the measuring unit is a chamber for measuring the liquid, and the liquid provided at one end thereof. An opening made up of a measuring unit body having an introduction port for introducing the liquid and a discharge port for discharging the liquid provided at the other end, and a through-hole penetrating in the thickness direction or a groove provided on the first substrate surface And a connection flow path connecting the discharge port and the opening. Here, the measuring unit main body and the opening are arranged so as to face each other with the connection channel interposed therebetween.

  In the microchip of the present invention, the depth L of the weighing unit body is preferably larger than the depth M of the connection channel, and the ratio L / M of the depth L of the measurement unit body to the depth M of the connection channel is More preferably, it is in the range of 2/1 to 9/1. Moreover, it is more preferable that the bottom surface of the measuring unit main body and the bottom surface of the connection channel are connected by an inclined surface.

  Further, in the microchip of the present invention, of the side walls of the grooves or the through holes constituting the opening, the side wall continuous to the bottom of the connection channel has an inner angle formed by the side wall and the bottom of the connection channel. It is preferable to incline with respect to the thickness direction of the first substrate so as to be less than 90 degrees.

  Moreover, it is preferable that the connection channel and the opening are connected so that the connection surface has an angle with respect to the liquid level in the connection channel of the measured liquid.

  Furthermore, in the microchip of the present invention, it is preferable that an introduction flow path for introducing a liquid to the introduction port is connected to the introduction port of the measuring unit main body. In this case, the depth N of the introduction channel is more preferably smaller than the depth L of the measuring unit main body. More preferably, the bottom surface of the measuring unit main body and the bottom surface of the introduction channel are connected by an inclined surface.

  It is preferable that the measuring unit main body has a substantially triangular shape when viewed from the stacking direction of the substrates. In this case, it is more preferable that the measuring unit main body includes an introduction port at one corner of the substantially triangular shape and a discharge port at the other corner.

  The microchip of the present invention includes a first substrate provided with grooves formed on both surfaces and / or through holes penetrating in the thickness direction, and a second substrate and a third substrate sandwiching the first substrate. And a first fluid circuit comprising a cavity formed by a groove provided on one surface of the first substrate and a first substrate side surface of the second substrate, and in the first substrate The microchip which has the 2nd fluid circuit which consists of the cavity provided by the groove | channel provided in the other surface and the 1st board | substrate side surface in a 3rd board | substrate may be sufficient. In this case, it is preferable that the opening is formed of a through hole that connects the first fluid circuit and the second fluid circuit and penetrates the first substrate in the thickness direction.

  ADVANTAGE OF THE INVENTION According to this invention, the microchip with which the occupation space of the measurement part was reduced can be provided. By reducing the space occupied by the measuring unit, it is possible to further reduce the size of the microchip and the restriction on the fluid circuit structure design. In addition, according to the present invention, it is possible to provide a microchip including a measuring unit with a small amount of measurement error, and thereby it is possible to accurately measure a liquid. Measuring liquid with high accuracy brings about improvement in accuracy and reliability of inspection / analysis using a microchip.

  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 preferable form of the present invention, the microchip is composed of a first substrate and a second substrate laminated and bonded on the first substrate, and more specifically, a groove and a surface are formed on the surface. The second substrate is bonded to the first substrate having a through hole penetrating in the thickness direction so that the groove forming surface of the first substrate faces the second substrate. Therefore, the microchip composed of the two substrates has a hollow portion formed therein with a groove provided on the surface of the first substrate and a surface of the second substrate on the side facing the first substrate. A fluid circuit comprising: The shape and pattern of the groove formed on the surface of the first substrate are not particularly limited, but an appropriate fluid circuit in which the structure of the cavity constituted by the groove and the surface of the second substrate is desired. It is determined to be a structure.

  In another preferred embodiment of the present invention, the microchip sandwiches the first substrate having a groove provided on both surfaces of the substrate and / or a through hole penetrating in the thickness direction, and the first substrate. It consists of the laminated | stacked and bonded 2nd board | substrate and 3rd board | substrate. 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 first fluid circuit and the second fluid circuit can be connected by one or two or more through-holes penetrating in the thickness direction formed in the first substrate.

  Here, 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 an adhesive is used. The method of using and adhering can be mentioned. 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 first substrate having a groove on the surface can be, for example, a transparent substrate. As a result, a detection unit composed of a transparent first substrate groove and a second substrate surface can be formed as a part of the fluid circuit, and the detection unit is an object of inspection and analysis. Optical measurement such as introducing a mixed liquid of a specimen and a liquid reagent, irradiating the detection unit with light, and detecting the intensity (transmittance) of the transmitted light can be performed on the mixed liquid. . The second 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 second substrate as a colored substrate, a welding method using light such as a laser can be used. Further, when the substrates are bonded by the laser welding method, the bonded surface of the colored substrate is mainly melted and bonded, so that the deformation of the groove formed on the transparent substrate that is the first substrate is changed. Can be minimized.

  Further, when the microchip is constituted by three sheets of the first substrate, the second substrate, and the third substrate, for example, a first provided with a groove formed on both surfaces and / or a through hole penetrating in the thickness direction. The second substrate and the third substrate that sandwich the substrate may be transparent substrates. Thereby, as a part of the fluid circuit, it is possible to form a detection unit composed of a through hole penetrating the first substrate in the thickness direction and the transparent second and third substrate surfaces, A liquid mixture of a specimen and a liquid reagent to be inspected / analyzed is introduced into the detection unit, and light in a direction perpendicular to the microchip surface is irradiated from the upper surface (or lower surface) side of the microchip to the detection unit. Optical measurement such as detecting the intensity (transmittance) of light transmitted from the opposite side 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 a groove (flow path pattern) constituting a fluid circuit on the surface of the first substrate is not particularly limited, and examples include an injection molding method using a mold having a transfer structure, an imprint method, and the like. Can do. 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.

  In the microchip of the present invention, the fluid circuit includes at least a measuring unit for measuring a sample such as a sample to be tested and analyzed and a liquid reagent mixed with the sample. The fluid circuit may include only one measuring unit or may include two or more measuring units. The measuring unit according to the present invention will be described in detail later.

  The fluid circuit may have a part other than the measuring part. Examples of such parts include a separation unit for taking out a specific component from a sample introduced into the fluid circuit; a liquid reagent holding unit for storing a liquid reagent; and a mixture for mixing the sample and the liquid reagent. Part; a detection part (for example, a cuvette for optical measurement) for performing inspection and analysis (for example, detection or quantification of a specific component in the liquid mixture) of the liquid mixture obtained by mixing the specimen and the liquid reagent Can be mentioned. 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. A plurality of these parts may be provided.

  In this specification, the “liquid reagent” is a liquid substance to be mixed with a specimen to be examined / analyzed, and the specimen is processed in the examination / analysis using a microchip, or the specimen It is a liquid substance for reacting with. Only one type of liquid reagent may be incorporated in one microchip, or two or more types of liquid reagents may be incorporated. Further, in this specification, “specimen” refers to a substance (for example, blood) to be tested / analyzed introduced into a fluid circuit itself or a specific component (for example, a plasma component, a blood cell component, etc.) in the substance. ).

  In the case where the microchip is composed of two substrates (a first substrate and a second substrate), the microchip of the present invention usually has a surface of the first substrate (this surface is usually used as a microchip). A liquid for injecting the liquid reagent into the liquid reagent holding part, which is a through-hole penetrating to the internal liquid reagent holding part (through the first substrate in the thickness direction). A reagent 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 (first substrate surface) after the liquid reagent is injected from the liquid reagent inlet. Is affixed for use. When the microchip is composed of three substrates (first substrate, second substrate, and third substrate), the liquid reagent injection port has the second substrate or the third substrate in the thickness direction. It can be provided as a through-hole penetrating through.

  The liquid mixture finally obtained by mixing the specimen and one or more liquid reagents is not particularly limited. For example, light is applied to a site (for example, a detection unit) in which the liquid mixture is stored. It is used for optical measurement such as a method for detecting the intensity (transmittance) of light transmitted through irradiation, 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 includes a rotor (rotor) that is rotatable about a centrifugal axis, and a rotatable stage that is disposed on the rotor. Place the microchip on the stage, rotate the stage to arbitrarily set the angle of the microchip with respect to the rotor, and rotate the rotor around the centrifugal axis to move the microchip in any direction. Centrifugal force can be applied.

  Hereinafter, the measuring unit provided in the microchip of the present invention will be described in detail. In the following, a description will be given mainly using a microchip composed of a first substrate and a second substrate and a third substrate sandwiching the first substrate as an example. However, as described above, the microchip of the present invention is not limited to this. It is not limited to this, and it may be composed of two substrates.

  FIG. 3 is a top view showing an example of the structure of the weighing unit provided in the microchip of the present invention. More specifically, FIG. 3 is a top view showing an example of the groove shape of the first substrate 201 forming the measuring portion included in the microchip of the present invention, and the second stacked on the first substrate 201. The substrate 202 and the third substrate 203 are omitted in FIG. FIG. 4 is a schematic cross-sectional view taken along the line III-III ′ shown in FIG. 3 (the second substrate 202 and the third substrate 203 are shown in FIG. 4). The weighing unit 100 shown in FIGS. 3 and 4 includes a weighing unit main body 101 that is a chamber for measuring a liquid and has a volume approximately equal to the amount of the liquid to be measured, and a first substrate 201. And an opening 102 formed of a through hole penetrating in the thickness direction. One end of the measuring unit main body 101 has an introduction port 103 for introducing a liquid to be weighed, and the other end has an excess liquid (waste liquid) exceeding the capacity of the measuring unit main body 101. Has a discharge port 104 for discharging the water. The discharge port 104 and the opening 102 are connected by a connection channel 105 that is a linear channel. The introduction port 103 is connected to an introduction channel 106 for guiding the liquid to be measured to the introduction port 103.

  In the measuring unit 100 shown in FIGS. 3 and 4, the measuring unit main body 101, the connection channel 105 and the introduction channel 106 are sandwiched between the first wall 110 and the second wall 120 having a V-shaped part. The bottom surfaces of the measuring unit main body 101, the connection channel 105, and the introduction channel 106 are part of the bottom surface of the groove provided on the surface of the first substrate 201.

  In the measuring unit 100 according to the present invention, as shown in FIGS. 3 and 4, the measuring unit main body 101 and the opening 102 formed of a through hole penetrating the surface of the first substrate 201 in the thickness direction are connected. It arrange | positions so that the flow path 105 may be pinched | interposed. The opening 102 has a function of causing an excessive amount of liquid (waste liquid) overflowing from the measuring unit main body 101 when the liquid is introduced into the measuring unit main body 101 to flow out to the “other part” in the fluid circuit. In the microchip shown in FIG. 3 and FIG. 4, “other part” means a waste liquid reservoir formed in the second fluid circuit formed by the first substrate 201 and the third substrate 203. is there.

  For example, in the metering unit shown in FIGS. 1 and 2 described above, when the waste liquid is introduced into the metering unit main body with respect to the discharge port, the waste liquid can be stored in the waste liquid reservoir. It is necessary to dispose downstream in the centrifugal force direction, and for this reason, the waste liquid disposed at the downstream side in the centrifugal force direction by curving the discharge flow path connecting the discharge port and the waste liquid reservoir. It must be formed to extend to the reservoir. When such a structure is adopted, a problem arises in that the space occupied by the measuring portion increases due to the long and curved discharge flow path. Such an increase in occupied space is not desirable for further miniaturization of the microchip, and may limit the degree of freedom in designing the fluid circuit structure. On the other hand, when the measuring unit main body and the opening for discharging the waste liquid are arranged so as to face each other through the connection flow path as in the measuring unit according to the present invention, there is a restriction on the place where the waste liquid reservoir is arranged. At the same time, there is no need to bend or lengthen the connection channel. As a result, the space occupied by the weighing unit can be reduced. In addition, since the liquid (waste liquid) discharged from the measuring unit main body passes through the connection channel and then drops in the thickness direction of the first substrate from the opening, the liquid is introduced into the measuring unit main body. Immediately after the application of the centrifugal force is stopped, the liquid in the measuring unit main body and the connection flow path (measured liquid) and the liquid flowing out to the opening (waste liquid) become good, and measurement errors are less likely to occur. Accurate metering of liquid can be performed.

  Here, referring to FIG. 4, the depth L of the measuring unit main body 101 is preferably larger than the depth M of the connection flow path 105. The liquid is introduced into the measuring unit main body 101 by applying a centrifugal force to the microchip, and after the liquid is measured by discharging excess liquid exceeding the capacity of the measuring unit main body 101 from the opening 102, the liquid is measured. When the application of the centrifugal force is stopped, the liquid level of the measured liquid is positioned in the connection flow path 105. At this time, if the depth M of the connection channel 105 is made smaller than the depth L of the measuring unit main body 101, the surface area of the liquid surface in the connection channel 105 becomes small. Differences are unlikely to occur and weighing errors can be further reduced.

  The ratio L / M of the depth L of the measuring unit main body 101 to the depth M of the connection channel 105 is preferably in the range of 2/1 to 9/1. When the ratio is smaller than 2/1, the volume of the measuring unit main body 101 is relatively small with respect to the volume of the connection channel 105, and as a result, the shape of the liquid level after measurement in the connection channel 105 The difference in the amount of liquid in the connection flow path 105 based on the difference between the two causes a relatively large influence on the total amount of the measured liquid, and the measurement error tends to increase. Further, if the ratio exceeds 9/1, it may be necessary to increase the thickness of the first substrate 201. Therefore, the ratio is 9 under the recent situation where miniaturization and thinning of the microchip are required. / 1 or less is preferable. Specifically, the depth M of the connection channel 105 is preferably about 0.2 to 0.5 mm, for example. If the depth M is less than 0.2 mm, the liquid in the connection flow path 105 may flow out to the opening 102 due to capillary action after measurement, which may cause a measurement error. In addition, when the depth M exceeds 0.5 mm, the surface area of the liquid surface in the connection flow path 105 becomes large, and measurement errors are likely to occur. Moreover, it is preferable that the width W1 (refer FIG. 3) of the connection flow path 105 shall be about 0.2-0.5 mm from the reason similar to the above.

  Note that the measuring unit main body 101 preferably has a capacity of about 50 to 150 times the volume of the connection channel 105. As a result, the influence of the difference in the liquid amount of the liquid in the connection channel 105 based on the difference in the shape of the liquid level after measurement in the connection channel 105 on the total amount of the measured liquid is reduced. Measurement error can be reduced.

  As described above, when the depth M of the connection channel 105 is made smaller than the depth L of the measurement unit main body 101, the bottom surface 101a of the measurement unit main body and the bottom surface 105a of the connection channel are formed by the inclined surface 107. It is preferable that they are connected (see FIG. 4). In this way, when the inclined surface is formed, when excess liquid (waste liquid) is discharged in the direction of the opening 102, air is accumulated in the corner formed by the inclined surface 107 and the bottom surface 101a of the measuring unit main body. Therefore, the measurement error can be suppressed and the liquid can be accurately measured. Referring to FIG. 4, angle θ formed between inclined surface 107 and bottom surface 101a of the measuring unit main body can be, for example, 90 ° <θ <180 °, and preferably 90 ° <θ ≦ 135 °.

  In the present invention, the shape of the measuring unit main body (the shape when viewed from the stacking direction of the substrates) is not particularly limited, but is preferably triangular or substantially triangular as shown in FIG. . In the measuring unit 100 shown in FIG. 3, such a shape is realized by a linear wall surface from the discharge port 104 of the first wall 110 to the introduction port 103 and a V-shaped wall surface of the second wall 120. Has been. As described above, when the measuring unit main body is configured using the V-shaped wall surface, when the liquid is introduced into the measuring unit main body, air hardly accumulates in the measuring unit main body, and the measuring unit main body is reliably filled with the liquid. Therefore, accurate weighing can be performed. In addition, the wall surface of the 2nd wall 120 is not limited to V shape, For example, a U shape may be sufficient. Further, the wall surface from the discharge port 104 to the introduction port 103 in the first wall 110 does not necessarily have to be linear, and may be V-shaped or U-shaped as in the second wall 120, for example. Good.

  In the case where the shape of the measuring unit main body is triangular or substantially triangular, the introduction port is disposed at one corner of the triangular shape and the discharge port is disposed at the other corner as in the weighing unit 100 of FIG. Is preferred. As a result, the entire space in the measuring unit main body is filled with the liquid.

  In the present invention, the shape of the connection channel is not particularly limited. However, from the viewpoint of ease of forming the flow path, it is preferable that the flow path is linear (having a square shape such as a square, a rectangle, or a trapezoid when viewed from the stacking direction of the substrates). . In addition, the length of the connection channel (distance from the outlet of the measuring unit main body to the opening) minimizes the amount of liquid to be measured that causes the liquid to be measured to be dragged and discharged by the waste liquid. In view of the point that it is preferable to keep the liquid surface (liquid running interface) formed in the connection channel and the measuring unit main body 101 as far as possible, it is preferable that the thickness is 1.0 mm or more. From the viewpoint of reducing the occupied space, the thickness is preferably 2.0 mm or less.

  Next, the opening 102 will be described in more detail. In the measuring unit 100 shown in FIGS. 3 and 4, the opening 102 includes a through hole that penetrates the first substrate 201 in the thickness direction. Of the liquid introduced into the measuring unit main body 101 by the application of the centrifugal force, the excess liquid (waste liquid) exceeding the capacity flows out to the opening 102 connected to the end of the connection channel 105, and the waste liquid is , And is accommodated in a second fluid circuit formed by the first substrate 201 and the third substrate 203. By such an operation, the liquid in the measuring unit main body 101, a part in the connection flow path 105, and a part in the introduction flow path 106 is filled, and the liquid is measured. As described above, the liquid that should remain in the connection channel 105 and the measurement unit main body 101 as the measured liquid and the waste liquid that should flow out to the opening 102 are sheared at the opening on the connection channel 105 side of the opening 102. Therefore, the liquid breakage between these liquids is good. That is, it is possible to effectively suppress the liquid that should remain in the connection flow path 105 and the measuring unit main body 101 from being dragged out by the waste liquid, and thus the measurement error can be reduced.

  Here, of the inner wall surface of the opening 102, the side wall surface 102a continuous to the bottom surface 105a of the connection flow path has an inner angle α formed by the side wall surface 102a and the bottom surface 105a of the connection flow path of less than 90 °. It is preferable that the first substrate 201 be inclined with respect to the thickness direction. By providing such an inclination, the shearing can be more effectively performed and the liquid breakage can be improved. In this case, the internal angle α can be set to about 80 ° ≦ α <90 °, for example. Further, not only the side wall surface 102a is inclined, but the through hole itself constituting the opening 102 is tapered such that the cross-sectional area of the opening increases as it advances from the connection flow path 105 side to the second fluid circuit side. You may have a shape. By making the taper shape, it is possible to more effectively prevent the liquid from touching the side wall surface and being dragged by surface tension or the like. Further, in order to effectively perform the shearing, the corner portion is formed as shown in FIG. 4 rather than the curved surface in the region where the side wall surface 102a and the bottom surface 105a of the connection channel intersect. preferable.

  The opening shape of the opening 102 shown in FIG. 3 (the shape when viewed from the stacking direction of the substrate) is a square shape, but is not limited to this, and for example, a circular shape, a polygonal shape, etc. Various shapes can be employed. However, as shown below, the opening shape of the opening includes the connection surface between the connection channel and the opening (the surface where the connection channel and the opening are connected), and the liquid level after the measurement. It is preferable to determine the relationship.

  FIG. 5 is an enlarged view showing the connection channel and the vicinity of the opening of the measuring unit, and is a schematic top view showing the relationship between the connection surface between the connection channel and the opening and the liquid level of the liquid after measurement FIG. The dotted line in FIG. 5 indicates the position of the liquid surface of the measured liquid when the application of the centrifugal force is stopped after the measurement. FIG. 5A shows the case of a weighing unit having the same structure as the weighing unit 100 shown in FIG. As shown in FIG. 5 (a), the orientation of the connection surface is adjusted so that the connection surface between the connection channel and the opening has an angle rather than parallel to the liquid level of the measured liquid. Since the contact between the liquid level and the opening is a point contact, the measured liquid and the waste liquid that should remain in the connection flow path and the main body of the measuring section are better disconnected, and the weighing error is reduced. It can be made smaller. On the other hand, when the opening shape of the opening is circular (FIG. 5B), or when the connection surface between the connection channel and the opening is parallel to the liquid level of the measured liquid (see FIG. 5). 5 (c)), the contact between the liquid surface and the opening is not a point contact (or is difficult to be a point contact), and the liquid breakage tends to be not as good as in the case of FIG. 5 (a). Therefore, it is preferable to connect the connection flow path and the opening so that the connection surface between the connection flow path and the opening has an angle with respect to the liquid level of the weighed liquid. It is preferable to adjust the shape and the like. The direction of the centrifugal force applied when the liquid is introduced into the measuring unit main body and the measurement is performed is the direction in which the liquid surface position is formed as shown in FIG. 5, that is, the entire space in the measuring unit main body and The direction in which a part in the connection channel and a part in the introduction channel are filled with the liquid is preferable.

  In FIG. 4, the opening 102 is a through hole that penetrates the first substrate 201 in the thickness direction, but the opening is not limited to this form in the present invention. For example, the opening may be a groove (concave portion) provided on the surface of the first substrate. In this case, the waste liquid is accommodated in the groove.

  Next, the introduction flow path 106 connected to the introduction port 103 will be described. The introduction channel 106 has a function of guiding the liquid to the introduction port 103 and also has a function of controlling the flow rate of the liquid introduced into the measuring unit main body. By restricting the flow rate of the liquid to an appropriate amount, it is possible to suppress or prevent air from being mixed with the liquid in the measuring unit main body. Here, when the depth N of the introduction flow path 106 is made smaller than the depth L of the measuring unit main body, the flow rate control function of the introduction flow path 106 can be further improved. In order to provide a good flow rate control function, it is preferable that the depth N and the width of the introduction channel 106 are about 0.2 to 0.5 mm, respectively. For the same reason as described above, it is preferable that the bottom surface 101a of the measuring unit main body and the bottom surface of the introduction channel 106 are connected by an inclined surface.

  In the present invention, as shown in FIG. 3, it is preferable to provide an air hole 130 in the measuring portion. By forming an air hole, a path for discharging the air that flows in together with the introduction of air or liquid existing in the measuring unit main body is secured, so the liquid measuring unit It is possible to smoothly move the liquid such as introduction into the main body and discharge of the waste liquid. The position of the air hole is not particularly limited. The air hole can be formed, for example, by forming a recess in a wall formed on the surface of the first substrate.

  EXAMPLES Hereinafter, although an Example is shown and this invention is demonstrated in detail, this invention is not limited to this.

  6 to 8 are plan views showing an example of the microchip of the present invention. The state of the liquid in the fluid circuit before use of the microchip and some steps of fluid processing performed using the microchip. The state of the liquid in is shown. 6-8, (a) shows the shape of a groove formed on one surface of the first substrate, and one fluid circuit (hereinafter referred to as “fluid circuit”) of the two-layer fluid circuits of the microchip. Also referred to as the upper fluid circuit.) The structure is shown. 6-8, (b) shows the shape of a groove formed on the other surface of the first substrate, and the other fluid circuit (hereinafter also referred to as the lower fluid circuit) of the microchip. .) Show the structure. The microchip 600 actually includes a second substrate and a third substrate that sandwich the first substrate, but is omitted in FIGS. Hereinafter, a part of the fluid processing operation of the microchip will be described with reference to FIGS.

  FIG. 6 is a plan view showing a state before fluid treatment (before use). As shown in FIG. 6, the microchip 600 includes liquid reagent holding units 601, 602, 603, 604, 605, and 606 for storing reagents mixed with plasma components or blood cell components in whole blood. The reagents R1, R2, R3, R4, R5, and R6 are preliminarily incorporated in these liquid reagent holders. A sample (whole blood) to be examined / analyzed is introduced into the fluid circuit from the sample inlet 607. The microchip 600 includes liquid reagent measuring units 701, 702, 703, 704, 705, and 706, which are measuring units according to the present invention. These measuring parts are parts for measuring the reagents R1, R2, R3, R4, R5 and R6, respectively.

  In a whole blood test using the microchip 600, first, whole blood collected from the sample inlet 607 is introduced. Next, referring to FIG. 7, a downward (downward in FIG. 7) centrifugal force is applied. Thereby, the whole blood is introduced into the separation unit 710 and centrifuged into a plasma component and a blood cell component (see FIG. 7B). Further, by this downward centrifugal force, each reagent moves to the lower fluid circuit through each of the through holes 801, 802, 803, 804, 805, 806 penetrating the first substrate in the thickness direction (FIG. 7 (b)).

  Next, referring to FIG. 8, a centrifugal force in the right direction (right direction in FIG. 8) is applied. Thereby, the plasma component in the separation part 710 moves to the area | region a (refer FIG.8 (b)). Reagents R1, R2, R3, R4, R5, and R6 that have moved to the lower fluid circuit, which is the fluid circuit on the side having the metering unit, are liquid reagent metering units 701, 702, 703, 704, 705 and It is introduced into 706 and weighed (see FIG. 8B). The details of the weighing operation are as described above. Each of these liquid reagent measuring units includes openings 701a, 702a, 703a, 704a, 705a, and 706a arranged so as to face the measuring unit main body with the connection channel interposed therebetween, and overflows from the connection channel. Excess reagent (waste liquid) moves through the opening to the upper fluid circuit and is accommodated (see FIG. 8A). Thus, according to the microchip 600 of the present embodiment, the reagents R1 to R6 can be simultaneously and accurately measured using the liquid reagent measuring unit. In addition, each liquid reagent measuring unit is designed to reduce the occupied area shown in the fluid circuit. Even after obtaining the state shown in FIG. 8, by applying a centrifugal force in an appropriate direction, fluid processing such as measurement of the plasma component and blood cell component, mixing of the plasma component or blood cell component and each reagent, etc. It is done, but I will omit it here.

  It should be understood that the embodiments and examples disclosed herein are illustrative and non-restrictive in every respect. 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 top view which shows an example of the structure of the measurement part with which a microchip is provided. It is a top view which shows another example of the structure of the measurement part with which a microchip is provided. It is a top view which shows an example of the structure of the measurement part with which the microchip of this invention is provided. It is a schematic sectional drawing in the III-III 'line | wire shown by FIG. It is a schematic top view which shows the relationship between the connection surface of a connection flow path and an opening part, and the liquid level of the liquid after measurement. It is a top view which shows an example of the microchip of this invention, and is a top view which shows the state before performing a fluid process (before use). It is a top view which shows the state of the liquid in one process of the fluid processing performed using the microchip shown by FIG. It is a top view which shows the state of the liquid in one process of the fluid processing performed using the microchip shown by FIG.

Explanation of symbols

  10, 60, 100 Weighing unit, 20, 70, 101 Weighing unit body, 30 opening, 40, 80 Discharge flow path, 50, 90 Waste liquid reservoir, 71 Inlet port, 72 discharge port, 101a Bottom surface of weighing unit body, 102 Opening, 102a Side wall surface continuous to bottom surface of connection flow path, 103 introduction port, 104 discharge port, 105 connection flow path, 105a bottom surface of connection flow path, 106 introduction flow path, 107 inclined surface, 110 first wall, 120 2nd wall, 130 air hole, 201 1st board | substrate, 202 2nd board | substrate, 203 3rd board | substrate, 600 microchip, 601,602,603,604,605,606 liquid reagent holding | maintenance part, 607 sample injection port 701, 702, 703, 704, 705, 706 Liquid reagent metering unit, 701a, 702a, 703a, 704a, 705a, 706a Open Department, 710 separation unit, 801,802,803,804,805,806 through hole.

Claims (10)

A first substrate having a groove and / or a through-hole penetrating in the thickness direction on the surface; and a second substrate laminated on the first substrate;
A microchip having a first fluid circuit composed of a cavity formed by the groove and the first substrate side surface of the second substrate,
The first fluid circuit includes a metering unit for metering a liquid,
The weighing unit is
A chamber for measuring the liquid, and a measuring unit main body provided with an inlet for introducing the liquid provided at one end thereof and an outlet for discharging the liquid provided at the other end; ,
An opening formed of a groove provided in the first substrate surface or a through hole penetrating in the thickness direction;
A connecting flow path connecting the outlet and the opening;
Have
The measuring part main body and the opening are microchips arranged so as to face each other with the connection channel interposed therebetween.   2. The microchip according to claim 1, wherein a depth L of the measuring unit main body is greater than a depth M of the connection channel.   3. The microchip according to claim 2, wherein a ratio L / M of a depth L of the measuring unit main body to a depth M of the connection channel is in a range of 2/1 to 9/1.   4. The microchip according to claim 2, wherein a bottom surface of the measuring unit main body and a bottom surface of the connection channel are connected by an inclined surface.   Of the side wall surfaces of the grooves or through holes that form the opening, the side wall surface that is continuous with the bottom surface of the connection channel has an inner angle formed by the side wall surface and the bottom surface of the connection channel that is less than 90 degrees. The microchip according to claim 1, wherein the microchip is inclined with respect to the thickness direction of the first substrate.   The said connection flow path and the said opening part are connected so that the connection surface may have an angle with respect to the liquid level in the said connection flow path of the said measured liquid. The microchip described. An introduction flow path for guiding the liquid to the introduction port is connected to the introduction port,
The depth N of the introduction channel is a microchip according to any one of claims 1 to 6, which is smaller than a depth L of the measuring unit main body.   The microchip according to claim 7, wherein a bottom surface of the measuring unit main body and a bottom surface of the introduction channel are connected by an inclined surface. The measuring unit body has a substantially triangular shape,
The microchip according to claim 1, wherein the introduction port is provided at one corner of the substantially triangular shape, and the discharge port is provided at the other corner. A first substrate provided with grooves formed on both surfaces and / or a through-hole penetrating in the thickness direction, and a second substrate and a third substrate sandwiching the first substrate,
A first fluid circuit comprising a cavity formed by a groove provided on one surface of the first substrate and the first substrate side surface of the second substrate; and A second fluid circuit composed of a cavity formed by a groove provided on the other surface and the first substrate side surface of the third substrate;
The micro according to any one of claims 1 to 9, wherein the opening includes a through hole that connects the first fluid circuit and the second fluid circuit and penetrates the first substrate in a thickness direction. Chip.

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