WO2019146102A1

WO2019146102A1 - Fluid device and use thereof

Info

Publication number: WO2019146102A1
Application number: PCT/JP2018/002692
Authority: WO
Inventors: 耕磨林; 陽介藤次; 太郎上野; 遼小林; 永波李
Original assignee: 株式会社ニコン
Priority date: 2018-01-29
Filing date: 2018-01-29
Publication date: 2019-08-01
Also published as: JPWO2019146102A1

Abstract

This fluid device is provided with a flow path, a first compartment valve and a second compartment valve which are arranged in the flow path and which define a prescribed region of the flow path by closing, and a first diaphragm member and a second diaphragm member which are arranged between the first compartment valve and the second compartment valve and which form at least a portion of a flow path wall in the flow path. In a state in which the first compartment valve and the second compartment valve are closed, at least one of the first diaphragm member and the second diaphragm member is deformable.

Description

Fluid device and use thereof

The present invention relates to fluidic devices and their use. More specifically, the present invention relates to a fluid device and a method of stirring a fluid.

In recent years, the development of μ-TAS (Micro-Total Analysis Systems) aimed at speeding up, increasing efficiency and integration of tests in the field of in vitro diagnostics and biological research, and ultraminiaturization of testing equipment, etc. Research is actively pursued worldwide.

The μ-TAS is superior to conventional testing instruments in that it can be measured and analyzed with a small amount of sample, can be carried, can be used at low cost, and can be disposable. In particular, when using an expensive reagent or when testing a small amount of many samples, it is noted as a highly useful method.

When performing enzyme reaction, antigen-antibody reaction, nucleic acid hybridization reaction, etc. in a small fluid device such as μ-TAS, the reaction solution is sufficiently agitated and detected from the viewpoint of improvement of reproducibility, enhancement of detection signal, etc. It is important to homogenize the reaction. Therefore, there is a need for a technique for stirring a small amount of solution and mixing the solution.

JP, 2014-77810, A

A fluidic device according to one embodiment includes a flow passage, a first partition valve and a second partition valve disposed in the flow passage, the first partition valve defining a predetermined region of the flow channel by closing, and the first A first diaphragm member and a second diaphragm member disposed between the first partition valve and the second partition valve and forming at least a part of the flow channel wall of the flow channel; The first diaphragm member and / or the second diaphragm member can be deformed with the compartment valve and the second compartment valve closed.

The fluidic device according to one embodiment includes a first substrate and a second substrate joined at a joining surface, and at least one of the first substrate and the second substrate flows by joining the two substrates. A groove forming a passage is provided on the joint surface, the groove includes two partition valves, and the first substrate is disposed between the two partition valves and positioned opposite to the groove The opening portion on the groove side of the first through hole has a through hole and a second through hole, and is closed by a first diaphragm member that can be deformed in a direction toward the axis of the flow path; The opening on the groove side of the through hole is closed by a second diaphragm member that is deformable in a direction away from the axis of the flow passage.

The method according to one embodiment includes a step of introducing the fluid into the flow channel of the fluid device, a step of closing the first partition valve and the second partition valve, and the first step. Deforming the diaphragm member in the direction toward the axis of the flow path and repeating the elimination of the deformation of the first diaphragm member.

It is a top view and a sectional view explaining the structure of fluid device 100 concerning one embodiment. (A) And (b) is sectional drawing explaining the structure of a diaphragm valve. (A) And (b) is sectional drawing explaining the structure of a diaphragm valve. (A) to (f) are schematic diagrams illustrating a method of stirring a fluid. (A) to (d) are schematic top views illustrating a method of quantifying and mixing two types of fluids. (A) to (d) are schematic views illustrating an analysis method according to an embodiment. (A) to (c) are photographs showing the results of Experimental Example 2. 7 is a graph showing the results of Experimental Example 3.

Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings as the case may be. In the drawings, the same or corresponding parts will be denoted by the same or corresponding reference symbols, without redundant description. In addition, the dimensional ratio in each figure has the part which exaggerates for description, and does not necessarily correspond with an actual dimensional ratio.

Fluid Device
First Embodiment
In one embodiment, the present invention provides a flow path, a first partition valve and a second partition valve disposed in the flow channel, which by closing define a predetermined area of the flow channel, and the first A first diaphragm member and a second diaphragm member disposed between the first partition valve and the second partition valve and forming at least a part of the flow channel wall of the flow channel; And a second diaphragm valve, wherein at least one of the first diaphragm member and the second diaphragm member can be deformed.

In the present fluid device, closing the first partition valve and the second partition valve defines an area including the first diaphragm member and the second diaphragm member of the flow passage, and the first The deformation of the second diaphragm member or the second diaphragm member stirs the fluid present in the defined area.

The volume of the defined area, that is, the volume of the fluid that can be agitated by the fluid device of the present embodiment is, for example, 10 μL or less, for example, 6 μL or less.

Conventionally, it has been difficult to agitate such minute volumes of fluid. In particular, in a fluid device such as the present invention, it has been difficult to agitate a minute volume of fluid over the flow path. Since the flow channel diameter of the flow channel into which a minute volume of fluid is introduced is designed to be thin, the fluid generally flows in a laminar flow in the flow channel. Therefore, different liquids and liquids of different densities introduced into the flow path are difficult to mix, and it takes a long time to stir so that the distribution becomes uniform.

On the other hand, as described later in the examples, the fluid device of this embodiment can efficiently agitate a minute volume of fluid. The fluid stirred in the fluid device of the present embodiment may be liquid or gas. In addition, when the liquid is stirred, the liquid may contain particles such as magnetic particles.

In the fluid device of the present embodiment, the fluid flow path means a flow path for fluid flow, and includes both the fluid flow state and the fluid non-flow state.

FIG. 1 is a top view and a cross-sectional view for explaining the structure of a fluid device 100 according to an embodiment. The fluid device 100 includes a flow path 110 for flowing the fluid L, a first partition valve 120, a second partition valve 130, a first diaphragm member 140, and a second diaphragm member 150. The first dividing valve 120, the first diaphragm member 140, the second diaphragm member 150, and the second dividing valve 130 are provided in the flow path 110 in this order.

The cross section of the flow channel 110 is, for example, substantially rectangular, circular, or elliptical. In the flow path 110, a part of each of the first diaphragm member 140, the second diaphragm member 150, the first partition valve 120, and the second partition valve 130 is exposed. By closing the first partition valve 120 and the second partition valve 130, a region 115 including the first diaphragm member 140 and the second diaphragm member 150 of the flow passage 110 is defined, and the first diaphragm member 140 or The deformation of the second diaphragm member 150 causes the fluid L present in the defined area 115 to be agitated.

In the example of FIG. 1, the first compartment valve 120 and the second compartment valve 130 are closed and the area 115 is defined. As used herein, "defined" refers to distinguishing the area of interest of the flow path from other areas of the flow path, and may be referred to as "shutoff", "independent", or the like.

The first diaphragm member 140 and the second diaphragm member 150 may be deformed in a direction perpendicular to the axial direction of the flow path 110 (the direction in which the fluid L flows). The direction perpendicular to the axial direction of the flow path 110 can be referred to as the direction toward the axis of the flow path 110, the thickness direction of the second substrate 220 described later, or the like.

A change in volume of the defined region 115 caused by the deformation of the second diaphragm member 150 or the first diaphragm member 140 due to the deformation of the first diaphragm member 140 or the second diaphragm member 150 Is absorbed.

More specifically, when the first diaphragm member 140 is deformed in a direction toward the axis of the flow passage 110 so as to project, the displaced fluid L moves the second diaphragm member 150 toward the outside of the flow passage 110. Push out and deform. That is, due to the deformation of the first diaphragm member 140, a pressure is applied to the fluid L from the first diaphragm member 140 toward the second diaphragm member 150, and the pressure causes the second diaphragm member 150 to be an axis of the flow passage 110. Transform in the direction away from the. Thereby, the volume of the region 115 is maintained at substantially the same volume as that before deformation of the first diaphragm member 140. Thus, when the first diaphragm member 140 or the second diaphragm member 150 is deformed, the second diaphragm member 150 or the first diaphragm member 140 is deformed to substantially change the volume of the region 115. Not to say that changes in volume are absorbed.

Similarly, when the second diaphragm member 150 deforms so as to project in the direction toward the axis of the flow passage 110, the displaced fluid L pushes the first diaphragm member 140 toward the outside of the flow passage to deform it. Let That is, due to the deformation of the second diaphragm member 150, a pressure is applied to the fluid L from the second diaphragm member 150 toward the first diaphragm member 140, and the pressure causes the first diaphragm member 140 to be an axis of the flow passage 110. Transform in the direction away from the. The volume of region 115 is maintained at substantially the same volume as before deformation of second diaphragm member 150.

From the deformation of the first diaphragm member 140 to the deformation of the second diaphragm member 150 or from the deformation of the second diaphragm member 150 to the deformation of the first diaphragm member 140, the fluid L present in the region 115 It may take some time before the influence of the change in volume of the region 115 is exerted. Therefore, there may be a time difference between the deformation of the first diaphragm member 140 and the deformation of the second diaphragm member 150 or the deformation of the second diaphragm member 150 and the deformation of the first diaphragm member 140. .

In the fluid device of this embodiment, as a result of deforming the first diaphragm member 140 in the direction toward the axis of the flow channel 110, the second diaphragm member 150 deforms in the direction away from the axis of the flow channel 110. May be Further, as a result of deforming the second diaphragm member 150 in the direction toward the axis of the flow passage 110, the first diaphragm member 140 may be deformed in a direction away from the axis of the flow passage 110. Further, as a result of deforming the first diaphragm member 140 so as to be recessed in a direction away from the axis of the flow path 110, a pressure is applied to the fluid L from the second diaphragm member 150 toward the first diaphragm member 140, and the pressure is The second diaphragm member 150 may be deformed in a direction toward the axis of the flow passage 110. Further, as a result of deforming the second diaphragm member 150 so as to be recessed in a direction away from the axis of the flow passage 110, a pressure from the first diaphragm member 140 toward the second diaphragm member 150 is applied to the fluid L. The first diaphragm member 140 may be deformed in a direction toward the axis of the flow passage 110.

In the example of FIG. 1, the first diaphragm member 140 is deformed in the direction toward the axis of the flow channel 110, and the second diaphragm member 150 is deformed in the direction away from the axis of the flow channel 110. Then, the deformation of the first diaphragm member 140 causes the change in volume of the region 115 to be absorbed by the deformation of the second diaphragm member 150, and the volume of the region 115 is the one before the deformation of the first diaphragm member 140. The same volume is maintained.

In the fluid device of the present embodiment, the amount of deformation of the first diaphragm member 140 and the amount of deformation of the second diaphragm member 150 may be substantially the same. In particular, when the shapes, sizes, materials, etc. of the first diaphragm member 140 and the second diaphragm member 150 are substantially the same, the amount of deformation of the first diaphragm member 140 and the amount of deformation of the second diaphragm member 150 are It tends to be substantially the same. Here, “substantially identical” means identical or nearly identical, and there is a difference on the order of errors that are difficult to avoid in the manufacture of fluidic devices.

In the fluid device of the present embodiment, the volume of the defined region 115 may be equal to the volume of the fluid L to be agitated. When the defined area 115 is completely filled with the fluid L, the volume of the defined area 115 and the volume of the fluid L to be agitated will be equal.

As described later, in the fluid device of the present embodiment, the first diaphragm member 140 and the second diaphragm member 150 of the flow path 110 are included by closing the first partition valve 120 and the second partition valve 130. Region 115 is defined. Also, with the region 115 defined, the first diaphragm member 140 or the second diaphragm member 150 deforms to cause the fluid L present in the defined region 115 to flow from the first diaphragm member 140. It moves in a direction toward the second diaphragm member 150 or in a direction from the second diaphragm member 150 toward the first diaphragm member 140 and is agitated.

Division valve
In the fluid device of the present embodiment, the first partition valve 120 and the second partition valve 130 are not particularly limited as long as they can define the region 115, but may be, for example, a diaphragm valve. The distance between the first partition valve 120 and the second partition valve 130 is, for example, about 0.1 to 100 mm, about 0.5 to 50 mm, and about 1 to 20 mm. The first partition valve 120 and the second partition valve 130 are disposed in the flow channel 110 and constitute a part of the flow channel 110. The fluid flowing through the flow path 110 contacts the first partition valve 120 and the second partition valve 130, and the flow of the fluid is changed by the opening and closing operation of the first partition valve 120 and the second partition valve 130.

The first partition valve 120 and the second partition valve 130 may operate so as to be able to open and close in the flow path 110. Unlike the diaphragm member described later, in particular, the surface opposite to the flow path may be in contact with the substrate so as not to deform in the direction away from the axis of the flow path, and control is performed so that the valve is deformed only in one direction It may be done.

FIGS. 2A and 2B are cross-sectional views for explaining an example of the structure of the diaphragm valve. 2A shows the diaphragm valve 200 in the open state, and FIG. 2B shows the diaphragm valve 200 in the closed state. As shown in FIGS. 2A and 2B, the diaphragm valve 200 includes a first substrate 210, an elastic member 230 made of an elastomeric material, and a second substrate 220. The second substrate 220 and the elastic member 230 are adhered in close contact with each other. Further, the space between the first substrate 210 and the diaphragm member 230 forms a flow path 110 for flowing the fluid L. In addition, a through hole 240 is provided in part of the second substrate 220. Further, in the through hole 240, the elastic member 230 is exposed.

In the open state of the diaphragm valve 200 shown in FIG. 2A, the fluid L can flow inside the flow path 110. On the other hand, as shown in FIG. 2B, when a fluid for controlling the valve is supplied from the through hole 240 of the diaphragm valve 200 and the inside of the through hole 240 is pressurized, the elastic member 230 is deformed and deformed. Part of the substrate 230 is in close contact with the first substrate 210. This state is a closed state of the diaphragm valve 200. As a result, the flow of the fluid L inside the flow path 110 is shut off.

In the example of FIG. 2B, the diaphragm valve 200 is deformed in a direction (direction toward the axis of the flow passage 110) perpendicular to the axial direction of the flow passage 110 (the direction in which the fluid L flows). Also, the diaphragm valve 200 is deformed in the thickness direction of the second substrate 220.

Here, as shown in FIGS. 2A and 2B, in order to make the closed state of the diaphragm valve 200 stronger, a convex portion is formed in the region of the first substrate 210 facing the through hole 240. 211 may be formed.

The diaphragm valve 200 may be controlled to open and close by fluid. Examples of the fluid for valve control include gases such as N ₂ gas and air, and liquids such as water and oil. The fluid for valve control can be supplied, for example, by a tube or the like connected to the through hole 240. Alternatively, opening and closing of the valve may be controlled by mechanical force or electromagnetic force.

The elastomer material forming the elastic member 230 is not particularly limited as long as it is a material that can be deformed in the axial direction of the through hole 240 according to the pressure change inside the through hole 240, for example, polydimethylsiloxane (PDMS) And silicone-based elastomers such as polymethylphenylsiloxane and polydiphenylsiloxane.

In the diaphragm valve 200 in the closed state shown in FIG. 2B, when the pressure applied to the inside of the through hole 240 is reduced, the elastic member 230 which has been deformed returns to the original shape, and again FIG. It will be in the open state shown in). As a result, the fluid L can flow again inside the flow path 110.

The structure of the diaphragm valve is not limited to the one described above. For example, the diaphragm valve 300 shown in FIGS. 3A and 3B can also be used. 3 (a) shows the diaphragm valve 300 in the open state, and FIG. 3 (b) shows the diaphragm valve 300 in the closed state.

As shown in FIGS. 3A and 3B, in the diaphragm valve 300, as compared with the diaphragm valve 200 described above, the elastic member 230 is not disposed on the entire surface of the second substrate 220. The points that are locally arranged only at the periphery are mainly different.

Since the elastic member 230 of the diaphragm valve 300 includes the anchor portion 231, the elastic member 230 may be broken and peeled off from the second substrate 220 even when the elastic member 230 is deformed by pressure. It is suppressed.

In the open state of the diaphragm valve 300 shown in FIG. 3A, the fluid L can flow inside the flow path 110. On the other hand, as shown in FIG. 3B, when the fluid for controlling the valve is supplied from the through hole 240 of the diaphragm valve 300 and the inside of the through hole 240 is pressurized, the elastic member 230 is deformed and deformed. Part of the substrate 230 is in close contact with the first substrate 210. This state is a closed state of the diaphragm valve 300. As a result, the flow of the fluid L inside the flow path 110 is shut off.

In the example of FIG. 3B, the diaphragm valve 300 is deformed in a direction (direction toward the axis of the flow path 110) perpendicular to the axial direction of the flow path 110 (the direction in which the fluid L flows). The diaphragm valve 300 is also deformed in the thickness direction of the second substrate 220.

In the diaphragm valve 300, as the fluid for valve control, the same one as the diaphragm valve 200 can be used. Alternatively, opening and closing of the diaphragm valve 300 may be controlled by mechanical force or electromagnetic force. Further, as the elastomer material forming the elastic member 230, the same one as the diaphragm valve 200 can be used.

In the diaphragm valve 300 in the closed state shown in FIG. 3B, when the pressure applied to the inside of the through hole 240 is reduced, the elastic member 230 which has been deformed returns to the original shape, and again FIG. It will be in the open state shown in). As a result, the fluid L can flow again inside the flow path 110.

When the first partition valve 120 and the second partition valve 130 are diaphragm valves, as shown in FIG. 2 (b) or FIG. 3 (b), the first partition valve 120 and the second partition valve 130 Closing may define a region 115 that includes the first diaphragm member 140 and the second diaphragm member 150 of the flow passage 110.

The first compartment valve 120 and the second compartment valve 130 may be normally closed valves or may be normally open valves. The normally closed valve is a valve that is closed in a steady state and is opened by operating the valve. In addition, the normally open valve is a valve that is open in a steady state and is closed by operating the valve.

When the first partition valve 120 and the second partition valve 130 are normally closed valves, in the steady state, the first partition valve 120 and the second partition valve 130 are in a closed state, and operate the valves. Thus, the region 115 defined by the first partition valve 120 and the second partition valve 130 is released.

Also, when the first partition valve 120 and the second partition valve 130 are normally open valves, in the steady state, the first partition valve 120 and the second partition valve 130 are open, and the valves are operated. As a result, the closed state is achieved, and a region 115 of a part of the flow channel 110 is defined.

<< diaphragm member >>
In the fluid device of the present embodiment, the first diaphragm member 140 and the second diaphragm member 150 may be formed of an elastomeric material. As the elastomeric material, the same one as described above for the elastic member of the diaphragm valve can be used. The first diaphragm member 140 and the second diaphragm member 150 may be controlled to open and close by fluid. Examples of the fluid for valve control include gases such as N ₂ gas and air, and liquids such as water and oil. Alternatively, opening and closing of the valve may be controlled by mechanical force or electromagnetic force.

The first diaphragm member 140 and the second diaphragm member 150 are disposed in the flow passage 110 and constitute a part of the flow passage wall of the flow passage 110. The fluid flowing through the flow path 110 contacts the first diaphragm member 140 and the second diaphragm member 150, and the deformation of the first diaphragm member 140 and the second diaphragm member 150 changes the flow of the fluid.

The first diaphragm member 140 and the second diaphragm member 150 are paired, and the other diaphragm member is deformed in the opposite direction to the one diaphragm member as the fluid is moved by the deformation of one diaphragm member. . Therefore, for example, the first diaphragm member 140 can be deformed in the direction toward the axis of the flow channel 110, and the second diaphragm member 150 can be deformed in the direction away from the axis of the flow channel 110. Alternatively, for example, the second diaphragm member 150 can be deformed in the direction toward the axis of the flow passage 110, and the first diaphragm member 140 can be deformed in the direction away from the axis of the flow passage 110. Alternatively, both the first diaphragm member 140 and the second diaphragm member 150 can be deformed in both the direction toward the axis of the flow passage 110 and the direction away from the axis of the flow passage 110. This point is different from the point that the first partition valve 120 and the second partition valve 130 described above may operate so as to be able to open and close in the flow path 110.

The first diaphragm member 140 and the second diaphragm member 150 may block the flow path 110 by deformation. Alternatively, the first diaphragm member 140 and the second diaphragm member 150 may not block the flow path 110 as long as the volume of the region 115 can be changed by deformation.

When the first diaphragm member 140 and the second diaphragm member 150 block the flow path 110 by deformation, the first diaphragm member 140 and the second diaphragm member 150 are the same as those described above. The diaphragm valve may be used. The first diaphragm member 140 and the second diaphragm member 150 may be the same as the first partition valve 120 and the second partition valve 130.

"Detection unit"
In the fluidic device of the present embodiment, the region 115 may be a reaction unit that performs an enzyme reaction, an antigen-antibody reaction, a nucleic acid hybridization reaction, and the like. As a result of stirring the fluid L present in the region 115, the above reaction is performed uniformly, and effects such as improvement of reproducibility and enhancement of a detection signal can be obtained.

The fluid device of the present embodiment may further include a detection unit that detects an enzyme reaction, an antigen-antibody reaction, a nucleic acid hybridization reaction, and the like. The detection unit is present outside the region 115, and the reaction solution after stirring may be sent to the detection unit to detect the reaction result. Alternatively, the detection unit may be present in the area 115.

For example, the detection unit may be disposed between the first diaphragm member 140 and the second diaphragm member 150 in the region 115.

When the detection unit is present in the area 115, detection may be performed in real time while stirring, that is, while performing an enzyme reaction, an antigen-antibody reaction, a nucleic acid hybridization reaction, etc. it can.

The detection unit may, for example, optically detect an enzyme reaction, an antigen-antibody reaction, a nucleic acid hybridization reaction or the like, or may electrically detect it. As a detector which detects optically, although a spectrophotometer, a fluorometer, etc. are mentioned, it is not limited to these.

Fluid Device
The fluidic device of the present embodiment includes a first substrate and a second substrate joined at a joining surface, and at least one of the first substrate and the second substrate is a flow path by joining the two substrates. A groove forming the joint surface, the groove comprising two partition valves, and the first substrate is a first through hole disposed between the two partition valves and in a position opposite to the groove The opening portion on the groove side of the first through hole has a hole and a second through hole, and is closed by a first diaphragm member which is deformable in a direction toward the axis of the flow passage, and the second through hole The groove side opening of the hole is closed by a second diaphragm member which is deformable in a direction away from the axis of the flow passage.

The fluidic device of the present embodiment may be a cartridge. That is, the fluid device of the present embodiment is a component that can be freely attached and detached, and may be attached to and detached from the analyzer. Alternatively, the fluidic device of the present embodiment may be part of a fluidic device incorporated in a cartridge that is attached to and detached from the analyzer.

In the fluid device of the present embodiment, at least two substrates have a stacked structure. A groove to be the flow path 110 is formed on the bonding surface of the two substrates. One substrate has at least two through holes, and a diaphragm member is provided on the bonding surface side of each of the through holes. These diaphragm members become the first diaphragm member 140 and the second diaphragm member 150. As described later, when the first partition valve 120 and the second partition valve 130 are diaphragm members, one substrate further includes two through holes, and the bonding surface side of each through hole is provided. An elastic member may be provided. These elastic members become the first partition valve 120 and the second partition valve 130.

[Method of stirring liquid]
First Embodiment
In one embodiment, the present invention is a method of agitating a fluid, comprising the steps of introducing the fluid into the flow path of the fluidic device described above, closing the first compartment valve and the second compartment valve. A method is provided, comprising the steps of: deforming the first diaphragm member in a direction toward the axis of the flow path, and repeating removing deformation of the first diaphragm member. According to the method of the present embodiment, a minute volume of fluid can be efficiently stirred.

The method according to the present embodiment is a method of stirring a fluid, wherein the step of introducing the fluid into the flow channel of the fluid device described above, the closing of the first partition valve and the second partition valve, Defining a region of the flow path including the first diaphragm member and the second diaphragm member, and deforming the first diaphragm member in a direction towards the axis of the flow passage, such that the definition Moving the fluid present in the selected area in a direction from the first diaphragm member toward the second diaphragm member, and deforming the second diaphragm member in a direction away from the axis of the flow passage; Eliminating the deformation of the first diaphragm member so that the fluid present in the region is directed from the second diaphragm member to the first diaphragm member Moving in a direction and removing deformation of the second diaphragm member, wherein the fluid present in the defined area is directed from the first diaphragm member to the second diaphragm member It may be a method of stirring by moving in the opposite direction or in the opposite direction.

Hereinafter, the fluid agitation method according to the first embodiment will be described with reference to FIGS. 4 (a) to 4 (d). FIG. 4A is a cross-sectional view showing an example of the fluid device described above. As shown in FIG. 4A, first, the fluid L is introduced into the flow path 110 of the fluid device 100. The method for introducing the fluid L into the flow channel 110 of the fluid device 100 is not particularly limited, and may be performed by a pump (not shown) or the like connected to the flow channel 110.

Subsequently, as shown in FIG. 4 (b), the first partition valve 120 and the second partition valve 130 are closed, and an area including the first diaphragm member 140 and the second diaphragm member 150 of the flow path 110. Define 115. When the area 115 is defined, the flow of the fluid L existing inside the area 115 is stopped and becomes stationary.

Subsequently, as shown in FIG. 4C, the first diaphragm member 140 is deformed in the direction toward the axis of the flow path 110 (the direction indicated by the arrow 140D1). The deformation of the diaphragm member 140 can be performed in the same manner as the control of the diaphragm valve described above. For example, the diaphragm member 140 can be deformed by supplying the same fluid as that described above as the valve control fluid in the direction of the arrow 140D1. Alternatively, the diaphragm member 140 may be deformed by mechanical force or electromagnetic force.

As a result, the fluid L present in the defined area 115 moves in the direction from the first diaphragm member 140 toward the second diaphragm member 150 (the direction indicated by the arrow LD1), and the second diaphragm member 150 flows. It deforms in the direction away from the axis of the path 110 (the direction indicated by the arrow 150D1).

The change in volume of the region 115 caused by the deformation of the first diaphragm member 140 is absorbed by the deformation of the second diaphragm member 150, and the volume of the region 115 is equal to that before the deformation of the first diaphragm member 140. The same volume is maintained.

Subsequently, as shown in FIG. 4D, the deformation of the first diaphragm member 140 is eliminated. The elimination of the deformation of the first diaphragm member 140 can be performed, for example, by stopping the supply of the valve control fluid that has been supplied in the direction of the arrow 140D1 in FIG. 4C.

As a result, the fluid L present in the region 115 moves in the direction from the second diaphragm member 150 toward the first diaphragm member 140 (the direction indicated by the arrow LD2), and the deformation of the second diaphragm member 150 is eliminated. Ru.

The change in volume of the region 115 caused by the elimination of the deformation of the first diaphragm member 140 is absorbed by the elimination of the deformation of the second diaphragm member 150, and the volume of the region 115 is the first diaphragm member 140. Is maintained in the same volume as before

The step of deforming the first diaphragm member 140 in the direction toward the axis of the flow passage 110 and the step of eliminating the deformation of the first diaphragm member 140 are preferably performed repeatedly several times.

Through the above-described steps, the fluid L present in the region 115 moves in the direction from the first diaphragm member 140 toward the second diaphragm member 150 (direction shown by arrow LD1) or in the opposite direction (direction shown by arrow LD2) It is stirred by.

In this manner, the fluid L present in the area 115 can be agitated by deforming the first diaphragm member 140 and restoring it.

Second Embodiment
The fluid agitation method according to the above-described embodiment further includes a step of repeating the step of deforming the second diaphragm member in the direction toward the axis of the flow path and eliminating the deformation of the second diaphragm member. The step of deforming the first diaphragm member to eliminate the deformation of the first diaphragm member, and the step of deforming the second diaphragm member to cancel the deformation of the second diaphragm member are alternated. May be performed.

For example, the second diaphragm member 150 is deformed in the direction toward the axis of the flow passage 110, so that the fluid L present in the defined region 115 is directed from the second diaphragm member 150 to the first diaphragm member 140. While moving in the direction (the direction indicated by the arrow LD2), the first diaphragm member 140 deforms in a direction away from the axis of the channel 110, and the deformation of the second diaphragm member 150 is eliminated. The fluid L present in 115 moves in a direction from the first diaphragm member 140 toward the second diaphragm member 150 (the direction indicated by the arrow LD1), and the deformation of the first diaphragm member 140 is eliminated; May be further provided.

While only the first diaphragm member 140 is deformed in the fluid agitation method according to the first embodiment, the fluid agitation method according to the second embodiment is not limited to the first diaphragm member 140, The second diaphragm member 150 also differs mainly in that it is deformed.

The fluid agitation method according to the second embodiment will be described with reference to FIGS. 4 (a) to 4 (f). In the fluid agitation method according to the second embodiment, the steps described above with reference to FIGS. 4A to 4D are the same as the fluid agitation method according to the first embodiment.

As described above with reference to FIG. 4D, when the deformation of the first diaphragm member 140 is eliminated, the direction of the fluid L present in the region 115 from the second diaphragm member 150 toward the first diaphragm member 140 While moving in the direction (indicated by the arrow LD2), the deformation of the second diaphragm member 150 is eliminated.

Subsequently, as shown in FIG. 4E, the second diaphragm member 150 is deformed in the direction toward the axis of the flow path 110 (the direction indicated by the arrow 150D2). The deformation of the diaphragm member 150 can be performed in the same manner as the deformation of the diaphragm member 140 described above.

As a result, the fluid L present in the defined area 115 moves in the direction from the second diaphragm member 150 toward the first diaphragm member 140 (the direction indicated by the arrow LD2), and the first diaphragm member 140 flows. It deforms in the direction away from the axis of path 110 (the direction indicated by arrow 140D2).

The change of the volume of the area 115 caused by the deformation of the second diaphragm member 150 is absorbed by the deformation of the first diaphragm member 140, and the volume of the area 115 is the same as that before the deformation of the second diaphragm member 150. The same volume is maintained.

Subsequently, as shown in FIG. 4 (f), the deformation of the second diaphragm member 150 is eliminated. The deformation of the second diaphragm member 150 can be eliminated, for example, by stopping the supply of the valve control fluid that has been supplied in the direction of the arrow 150D2 in FIG.

As a result, the fluid L present in the region 115 moves in the direction from the first diaphragm member 140 toward the second diaphragm member 150 (the direction indicated by the arrow LD1), and the deformation of the first diaphragm member 140 is eliminated. Ru.

The change in volume of the area 115 caused by the elimination of the deformation of the second diaphragm member 150 is absorbed by the elimination of the deformation of the first diaphragm member 140, and the volume of the area 115 is the second diaphragm member 150. Is maintained in the same volume as before

In this manner, the fluid L present in the region 115 is also agitated by deforming the first diaphragm member 140 and the second diaphragm member 150 in the direction toward the axis of the flow passage 110, respectively, and restoring them back. Can.

In the fluid agitation method according to the second embodiment, which of the first diaphragm member 140 and the second diaphragm member 150 is to be deformed first is not limited. In the example described above, the first diaphragm member 140 is deformed first, but the second diaphragm member 150 may be deformed first.

Third Embodiment
In the fluid agitation method according to the first embodiment and the second embodiment described above, the first diaphragm member is deformed in a direction away from the axis of the flow passage to eliminate the deformation of the first diaphragm member. And the first diaphragm member is deformed in a direction toward the axis of the flow channel to eliminate the deformation, and the first diaphragm member is moved away from the axis of the flow channel. The steps of repeating deformation and removing deformation of the first diaphragm member may be alternately performed.

For example, in the fluid agitation method according to the above-described first and second embodiments, the first diaphragm member 140 is deformed in the direction away from the axis of the flow passage 110 (the direction indicated by the arrow 140D2). While the fluid L present in the defined area 115 moves in the direction from the second diaphragm member 150 toward the first diaphragm member 140 (the direction indicated by the arrow LD 2), the second diaphragm member 150 The step of deformation in the direction toward the axis (the direction indicated by the arrow 150D2) and the deformation of the first diaphragm member 140 are eliminated. As a result, the fluid L present in the region 115 is removed from the first diaphragm member 140 While moving in the direction toward the diaphragm member 150 (the direction indicated by the arrow 150D1), the deformation of the second diaphragm member 150 is A step of erased, may further comprise a.

In the fluid agitation method according to the third embodiment, the direction and flow direction of the first diaphragm member 140 toward the axis of the flow passage 110 are compared with the fluid agitation method according to the first embodiment and the second embodiment. It mainly differs in that it deforms in both directions away from the axis of 110. In the fluid agitation method according to the first embodiment and the second embodiment, the first diaphragm member 140 is deformed only in the direction toward the axis of the flow passage 110.

Here, deforming means actively deforming the diaphragm member and does not include passive deformation of the diaphragm member to absorb changes in the volume of the region 115.

The fluid agitation method according to the third embodiment will be described with reference to FIGS. 4 (a) to 4 (f). The fluid agitation method according to the third embodiment is the same as the fluid agitation method according to the first and second embodiments in the steps described above with reference to FIGS. 4 (a) to 4 (d).

Subsequently, as shown in FIG. 4E, the first diaphragm member 140 is deformed in the direction away from the axis of the flow path 110 (the direction indicated by the arrow 140D2). The deformation of the diaphragm member 140 can be performed, for example, by sucking the diaphragm member 140 in the direction of the arrow 140D2. Alternatively, the diaphragm member 140 may be deformed by mechanical force or electromagnetic force.

As a result, the fluid L present in the defined area 115 moves in the direction from the second diaphragm member 150 toward the first diaphragm member 140 (the direction indicated by the arrow LD2), and the second diaphragm member 150 flows. It deforms in the direction towards the axis of the path 110.

In this process, the deformation direction of the diaphragm member 140, the deformation direction of the diaphragm member 150, and the movement direction of the fluid L are the same as those in the process described with reference to FIG. 4 (e) in the second embodiment. While the second diaphragm member 150 is actively deformed in the second embodiment, the third embodiment differs in that the first diaphragm member 140 is actively deformed.

The change in volume of the region 115 caused by the deformation of the first diaphragm member 140 is absorbed by the deformation of the second diaphragm member 150, and the volume of the region 115 is equal to that before the deformation of the first diaphragm member 140. The same volume is maintained. Here, the deformation of the second diaphragm member 150 is passive.

Subsequently, as shown in FIG. 4 (f), the deformation of the first diaphragm member 140 is eliminated. The elimination of the deformation of the first diaphragm member 140 can be performed, for example, by stopping the suction in the direction of the arrow 140D2 described above with reference to FIG.

As a result, the fluid L present in the region 115 moves in the direction from the first diaphragm member 140 toward the second diaphragm member 150 (the direction indicated by the arrow LD1), and the deformation of the second diaphragm member 150 is eliminated. Ru.

In this manner, the first diaphragm member 140 is deformed in the direction toward the axis of the flow path 110 and in the direction away from the axis of the flow path 110, and the fluid L present in the region 115 is also agitated by returning it. be able to.

Fourth Embodiment
In the fluid agitation method according to the first embodiment, the second embodiment, and the third embodiment described above, the second diaphragm member is deformed in a direction away from the axis of the flow passage, and the first diaphragm member The method further includes the step of repeating the elimination of the deformation, the step of deforming the first diaphragm member in the direction toward the axis of the flow path, and the step of eliminating the deformation; The method may be alternately performed in the steps of repeating deformation away from the axis and removing the deformation of the second diaphragm member.

For example, the second diaphragm member 150 is deformed in the direction away from the axis of the flow passage 110 (the direction indicated by the arrow 150D1), and as a result, the fluid L present in the defined area 115 is removed from the first diaphragm member 140 Moving in a direction (direction indicated by arrow LD1) toward the second diaphragm member 150 and deforming the first diaphragm member 140 in a direction (direction indicated by arrow 140D1) toward the axis of the flow path 110; As a result, the fluid L present in the region 115 moves in the direction (the direction indicated by the arrow LD2) from the second diaphragm member 150 toward the first diaphragm member 140, and And the step of eliminating the deformation of the one diaphragm member 140.

The fluid agitation method according to the fourth embodiment is different from the fluid agitation methods according to the first, second, and third embodiments in that the second diaphragm member 150 is used as the axis of the flow passage 110. It differs mainly in that it deforms in both directions, towards and away from the axis of the channel 110. In the fluid agitation method according to the first embodiment, the second embodiment, and the third embodiment, the second diaphragm member 150 is deformed only in the direction toward the axis of the flow path 110.

The fluid agitation method according to the fourth embodiment will be described with reference to FIGS. 4 (a) to 4 (f). The fluid agitation method according to the fourth embodiment includes the fluid agitation method according to the first embodiment, the second embodiment and the third embodiment in the steps described above with reference to FIGS. 4 (a) to 4 (b). Is the same as

As described above with reference to FIG. 4 (b), the first partition valve 120 and the second partition valve 130 are closed to include the first diaphragm member 140 and the second diaphragm member 150 of the flow passage 110. When the area 115 is defined, the flow of the fluid L existing inside the area 115 is stopped and becomes stationary.

Subsequently, as shown in FIG. 4C, the second diaphragm member 150 is deformed in the direction away from the axis of the flow path 110 (the direction indicated by the arrow 150D1). The deformation of the diaphragm member 150 can be performed, for example, by sucking the diaphragm member 150 in the direction of the arrow 150D1. Alternatively, the diaphragm member 150 may be deformed by mechanical force or electromagnetic force.

As a result, the fluid L present in the defined area 115 moves in the direction from the first diaphragm member 140 toward the second diaphragm member 150 (the direction indicated by the arrow LD1), and the first diaphragm member 140 flows. It deforms in the direction towards the axis of the path 110.

In this process, the deformation direction of the diaphragm member 150, the deformation direction of the diaphragm member 140, and the movement direction of the fluid L will be described with reference to FIG. 4C for the first embodiment, the second embodiment and the third embodiment. Are the same as in the While the second diaphragm member 150 is passively deformed in the first embodiment, the second embodiment and the third embodiment, the second diaphragm member 150 is actively deformed in the fourth embodiment. The point is different.

The change of the volume of the area 115 caused by the deformation of the second diaphragm member 150 is absorbed by the deformation of the first diaphragm member 140, and the volume of the area 115 is the same as that before the deformation of the second diaphragm member 150. The same volume is maintained. Here, the deformation of the first diaphragm member 140 is passive.

Subsequently, as shown in FIG. 4D, the deformation of the second diaphragm member 150 is eliminated. The deformation of the second diaphragm member 150 can be eliminated, for example, by stopping the suction in the direction of the arrow 150D1 described above with reference to FIG. 4C.

As a result, the fluid L present in the region 115 moves in the direction from the second diaphragm member 150 toward the first diaphragm member 140 (the direction indicated by the arrow LD2), and the deformation of the first diaphragm member 140 is eliminated. Ru.

Subsequently, as shown in FIG. 4E, the second diaphragm member 150 is deformed in the direction toward the axis of the flow path 110 (the direction indicated by the arrow 150D2). For example, the diaphragm member 150 can be deformed by supplying the same fluid as that described above as the valve control fluid in the direction of the arrow 150D2. Alternatively, the diaphragm member 150 may be deformed by mechanical force or electromagnetic force.

In this process, the deformation direction of the diaphragm member 140, the deformation direction of the diaphragm member 150, and the movement direction of the fluid L are the same as those in the process described with reference to FIG. 4 (e) in the third embodiment. In the third embodiment, the first diaphragm member 140 is actively deformed, whereas in the fourth embodiment, the second diaphragm member 150 is actively deformed.

Thus, the second diaphragm member 150 is deformed in the direction toward the axis of the flow path 110 and in the direction away from the axis of the flow path 110, and the fluid L present in the region 115 is also agitated by returning it. be able to.

The fluid agitation methods according to the first, second, third and fourth embodiments described above may be implemented in combination with one another.

For example, active deformation of the first diaphragm member 140 in the direction toward the axis of the flow passage 110 and in a direction away from the axis of the flow passage 110, and in the direction toward the axis of the flow passage 110 in the second diaphragm member 150 Active deformation of the flow path 110 away from the axis may be performed in combination. Alternatively, the movement of the fluid L in the region 115 in the direction indicated by the arrow LD1 and the direction indicated by the arrow LD2 is one cycle, and the first embodiment, the second embodiment, the third embodiment, and the fourth embodiment described above for each cycle You may implement, switching the stirring method of the fluid which concerns on form.

In the fluid agitation method according to the first, second, third, and fourth embodiments described above, the fluid L to be agitated may be a liquid or a gas. And granular materials. For example, the fluid L is a plurality of types of liquids, and may be mixed according to the stirring method of the present invention. Further, the fluid L may be a liquid or gas having uneven concentration, temperature, and distribution of components, and the distribution may be made uniform by the stirring method of the present invention. Further, the fluid L is a liquid in which the distribution state of the dispersion system such as an emulsion or the like is nonuniform, and the distribution state may be made uniform or finer by the stirring method of the present invention. The fluid L may contain particles such as magnetic particles, and the particles such as magnetic particles may be dispersed by the stirring method of the present invention.

As described later in the embodiment, even when the fluid L contains magnetic particles, the fluid L can be efficiently stirred. Further, since magnetic particles can be accumulated using magnetic force, for example, magnetic particles can be accumulated inside the region 115. Furthermore, the fluid L containing magnetic particles can be efficiently stirred by the method described above. Alternatively, for example, a magnet (not shown) may be brought close to the area 115 to exert a magnetic force, and magnetic particles may be accumulated in the area 115 in advance. Then, after the fluid L is introduced into the area 115 and the first dividing valve 120 and the second dividing valve 130 are closed to define the area 115, the magnet is released to release the magnetic particles in the area 115, The magnetic particles may be stirred into the fluid L according to the stirring method of

As a result, for example, an antibody, a nucleic acid fragment and the like can be bound to the magnetic particle, and an enzyme reaction, an antigen-antibody reaction, a nucleic acid hybridization reaction and the like can be performed on the magnetic particle.

Fifth Embodiment
The fluid agitation method according to the fifth embodiment is a method of quantifying and mixing two types of fluids. The fluid agitation method according to the fifth embodiment will be described with reference to FIGS. 5 (a) to 5 (d).

FIG. 5A is a schematic top view illustrating the structure of the fluid device 500 according to an embodiment. The fluid device 500 includes the

flow paths

110a, 110b, 110c, 110d for flowing fluid, the first partition valve 120, the second partition valve 130, the third partition valve 510, and the fourth partition valve. 520, a fifth dividing valve 530, a first diaphragm member 140, and a second diaphragm member 150.

Hereinafter, a procedure of quantifying and mixing the first fluid L1 and the second fluid L2 using the fluid device 500 will be described.

First, as shown in FIG. 5A, the third partition valve 510 is closed, and the first partition valve 120 and the fourth partition valve 520 are opened. Subsequently, the first fluid L1 is allowed to flow through the flow path 110a and the flow path 110b. Subsequently, as shown in FIG. 5B, the first partition valve 120 and the fourth partition valve 520 are closed. Thereby, in the flow path 110a, a region 115a surrounded by the first partition valve 120, the third partition valve 510, and the fourth partition valve 520 is defined, and the fluid L1 present in the region 115a is quantified. .

On the other hand, as shown in FIG. 5 (a) or (b), the third partition valve 510 is closed, and the second partition valve 130 and the fifth partition valve 530 are opened. Subsequently, the second fluid L2 is allowed to flow through the flow path 110c and the flow path 110d. Subsequently, as shown in FIG. 5C, the second partition valve 130 and the fifth partition valve 530 are closed. Thereby, in the flow path 110c, a region 115b surrounded by the second partition valve 130, the third partition valve 510, and the fifth partition valve 530 is defined, and the fluid L2 present in the region 115b is quantified. .

Subsequently, as shown in FIG. 5 (d), the third partition valve 510 is opened. As a result, the flow path 110 a and the flow path 110 b are connected, and the region 115 including the first diaphragm member 140 and the second diaphragm member 150 is defined by the first partition valve 120 and the second partition valve 130. It becomes a state. Region 115 includes fluid L1 and fluid L2 respectively quantified.

Subsequently, by deforming the first diaphragm member 140 or the second diaphragm member 150, the fluid L1 and the fluid L2 present in the defined area 115 are agitated and mixed. Here, how to deform the first diaphragm member 140 and the second diaphragm member 150 is the same as the fluid agitation method according to the first to fourth embodiments described above.

By the above operation, the fluid L1 and the fluid L2 can be quantified and mixed. Here, for example, a detector may be provided in the region 115. In this case, during or after stirring of the fluid L1 and the fluid L2, the state of the fluid in the area 115 can be detected and measured using the detector.

[Analytical method]
In one embodiment, the present invention provides an analysis method comprising the steps of stirring a fluid according to the method described above and analyzing the stirred fluid.

Hereinafter, with reference to FIGS. 6 (a) to 6 (d), a specific example of carrying out an antigen-antibody reaction according to the analysis method of the present embodiment will be described.

First, as shown in FIG. 6A, the fluid L is introduced into the flow path 110 of the fluid device 100. Here, the fluid L contains magnetic particles M. On the surface of the magnetic particle M, an antibody against a protein to be measured is bound.

Further, as shown in FIG. 6A, by setting the fluid device 100 to the magnetic stand 160, the magnetic particles M in the fluid L can be accumulated inside the region 115. Here, the area 115 is an area defined by closing the first partition valve 120 and the second partition valve 130.

Subsequently, as shown in FIG. 6B, the buffer LB for washing may be made to flow in the flow path 110 of the fluid device 100 to wash the magnetic particles M.

Subsequently, as shown in FIG. 6B, the sample containing the sample T is allowed to flow in the flow path 110 of the fluid device 100. The sample T is a detection target molecule to which an antibody bound to the surface of the magnetic particle M specifically binds.

Subsequently, as shown in FIG. 6 (c), the first partition valve 120 and the second partition valve 130 are closed to define the region 115, and the fluid device 100 is removed from the magnetic stand 160. As a result, the magnetic particle M is isolated from the sample containing the sample T inside the region 115. Also, here, the

diaphragm members

140 and 150 of the fluid device 100 are activated to stir the fluid L (the liquid containing the sample T and the magnetic particles M) inside the region 115 as described above. As a result, the fluid L in the region 115 becomes uniform, and the antibody disposed on the surface of the magnetic particle M and the sample T react efficiently with good reproducibility.

Subsequently, as shown in FIG. 6 (b), the fluid device 100 is set on the magnetic stand 160, and the magnetic particles M present in the region 115 are accumulated. Further, as shown in FIG. 6B, the washing buffer LB is caused to flow in the flow channel 110 to wash the magnetic particles M.

Subsequently, as shown in FIG. 6 (b), a buffer containing a secondary antibody (2nd Ab) is allowed to flow through the flow channel 110 of the fluidic device 100. The secondary antibody is a labeled antibody that specifically binds to sample T.

Subsequently, as shown in FIG. 6 (c), the first partition valve 120 and the second partition valve 130 are closed to define the region 115, and the fluid device 100 is removed from the magnetic stand 160. As a result, the buffer containing the secondary antibody and the magnetic particles M are isolated inside the region 115. Also, here, the

diaphragm members

140 and 150 of the fluid device 100 are activated to stir the fluid L (a liquid containing the magnetic particles M to which the secondary antibody and the sample T are bound) inside the region 115 as described above. . As a result, the fluid L inside the region 115 becomes uniform, and the sample T bound to the surface of the magnetic particle M and the secondary antibody react efficiently with good reproducibility.

Subsequently, as shown in FIG. 6B, the substrate reaction solution (S) corresponding to the label of the secondary antibody is allowed to flow through the flow channel 110 of the fluid device 100.

Subsequently, as shown in FIG. 6 (d), the first partition valve 120 and the second partition valve 130 are closed to define the region 115, and the fluid device 100 is removed from the magnetic stand 160. As a result, the substrate reaction solution S and the magnetic particles M are isolated in the region 115. Also, here, the

diaphragm members

140 and 150 of the fluid device 100 are activated, and the fluid L inside the region 115 (a fluid containing the substrate reaction solution S and the magnetic particles M bound to the secondary antibody) as described above is Stir. As a result, the fluid L in the region 115 becomes uniform, and the secondary antibody bound to the surface of the magnetic particle M and the substrate reaction solution S react efficiently with good reproducibility.

As a result, a signal is generated. The signal is, for example, fluorescence. Subsequently, the generated signal is detected. For example, as shown in FIG. 6D, the detection of the signal may be detected using a detector 170 installed adjacent to the area 115, or a detector (not shown) installed outside the fluidic device. It may be detected using

When the detector is installed outside the fluidic device, the reaction liquid after reaction in the area 115 shown in FIG. 6D is sent to move to a position detectable by the detector and moved. It is good to detect the signal.

As mentioned above, although the specific example which implements an antigen antibody reaction by the analysis method of this embodiment was demonstrated, an enzyme reaction, a nucleic acid hybridization reaction, etc. can also be performed by the same method. In the above-described example, the reaction between the antibody and the sample T, the reaction between the sample T and the secondary antibody, and the reaction between the secondary antibody and the substrate reaction liquid S are performed using the fluid device 100. The analysis method of is not limited to this, for example, only the reaction between the secondary antibody and the substrate reaction solution S may be performed using the fluid device 100, and the other reactions may be performed using another fluid device or the like.

EXAMPLES The present invention will next be described in more detail by way of examples, which should not be construed as limiting the invention thereto.

[Experimental Example 1]
(Manufacturing of fluid devices)
A fluidic device shown schematically in FIGS. 1 (a) and (b) was manufactured. In the manufactured fluid device (hereinafter referred to as "device A"), the cross section of the channel 110 is substantially rectangular, the width of the channel 110 is 0.5 to 1.0 mm, and the height is 0.2 to It was 0.5 mm. The distance between the first compartment valve 120 and the second compartment valve was 5.5 mm.

The thickness of device A was about 5 mm. The first partition valve 120 and the second partition valve 130 are diaphragm valves, the material of the diaphragm member is urethane elastomer, the thickness of the diaphragm member is 300 μm, and the diameter of the diaphragm valve is 2 mm The

Further, the first diaphragm member 140 and the second diaphragm member 150 are diaphragm valves similar to the first partition valve 120 and the second partition valve 130, and the material of the diaphragm member is a urethane elastomer, and the diaphragm The thickness of the member was 300 μm and the diameter of the diaphragm valve was 2 mm.

[Experimental Example 2]
(Stirring of magnetic particles)
The fluorescent magnetic particles were stirred using the device A manufactured in Experimental Example 1. First, the fluorescent dye Cy5 (Dojindo Chemical Laboratory), which is a biotin-labeled fluorescent dye, is immobilized by avidin-biotin binding to magnetic particles (JSR Co., Ltd.) with a diameter of 3 μm on which streptavidin is modified on the surface. Magnetic particles were prepared. The prepared fluorescent magnetic particles have a TBST (0.05% Tween 20-Tris Buffered Saline) buffer containing 0.1% BSA (bovine serum albumin) to a final concentration of 0.01% (hereinafter referred to as “0.1% It suspended in "BSA-TBST".), And used for the following experiments.

Subsequently, a 3 mm × 3 mm × 5 mm neodymium magnet was set in the device A. Subsequently, 150 μL of the fluorescent magnetic particle suspension prepared as described above was introduced into the device A, and the fluorescent magnetic particles were accumulated in the region 115 by passing through the channel 110. Thereafter, 10 μL of 0.1% BSA-TBST was introduced to fill the channel. FIG. 7 (a) is a photomicrograph of device A after introducing fluorescent magnetic particles. The first partition valve 120 and the second partition valve 130 are closed and in a partitioned state.

Subsequently, device A was activated. Specifically, after the first partition valve 120 and the second partition valve 130 were closed, the first diaphragm member 140 and the second diaphragm member 150 were operated. The first diaphragm member 140 and the second diaphragm member 150 were alternately operated with a time difference of 300 ms. The operations of the first diaphragm member 140 and the second diaphragm member 150 were both performed in the direction toward the axis of the flow passage 110.

Here, one of the first diaphragm member 140 and the second diaphragm member 150 which has not been actuated is passively deformed in the direction of absorbing the volume change of the region 115.

FIG. 7 (b) is a photomicrograph of device A after stirring. As a result, as shown in FIG. 7 (b), it was revealed that the magnetic particles were uniformly stirred in a very short time (about 1 second).

FIG. 7C is a photograph showing the result of detection of fluorescence of Cy5 labeled with magnetic particles by observing the device A after stirring with a fluorescence microscope. As a result, it was also confirmed from the fluorescence micrograph that the magnetic particles were uniformly stirred.

[Experimental Example 3]
(Detection of antigen-antibody reaction)
Detection of antigen-antibody reaction was performed using Device A manufactured in Experimental Example 1. As an antigen, cardiac troponin I (cardiaic troponin I, cTnI) was used.

First, an anti-cTnI antibody (HyTest) was attached to a magnetic particle (JSR) with a diameter of 3 μm, the surface of which was carboxyl-modified, via the amino group of the antibody. Subsequently, the magnetic particles were diluted with 0.1% BSA-TBST so that the final concentration was 0.01 w / v%.

In addition, an alkaline phosphatase was labeled to an anti-cTnI antibody (HyTest) having an epitope different from that of the anti-cTnI antibody bound to the magnetic particles, using an alkaline phosphatase labeling kit (Dojindo Laboratories, Inc.).

Subsequently, 50 μL of the diluted magnetic particles described above, 50 μL of cTnI serially diluted with 0.1% BSA-TBST to 0, 10, 100, and 1000 pg / mL, and the antibody concentration of 2 μg / mL. 50 μL of the above-mentioned alkaline phosphatase-labeled anti-cTnI antibody diluted as above was mixed and reacted at 37 ° C. for 5 minutes to form an antigen-antibody complex on the magnetic particles.

Subsequently, it was washed three times with 200 μL of 0.1% BSA-TBST while capturing magnetic particles with a neodymium magnet. Subsequently, 150 μL of 0.1% BSA-TBST was added to the washed magnetic particles to resuspend the magnetic particles. Subsequently, the resuspended magnetic particles were sent to the channel 110 of the device A, and the magnetic particles were accumulated in the area 115.

Subsequently, 10 μL of CDP-star (Thermo Fisher Scientific), which is a chemiluminescent substrate of alkaline phosphatase, is introduced into the channel 110 of device A, and the first partition valve 120 and the second partition valve 130 are closed. Region 115 was determined, and the luminescence due to the decomposition reaction of CDP-star was detected in real time. As a detector, μPMT (Hamamatsu Photonics) was used. The measurement was performed with the detector placed with the sensor adjacent to the area 115 of device A.

Three minutes after the start of the measurement, the first diaphragm member 140 and the second diaphragm member 150 were operated and agitation was performed for 90 seconds. The operation of the first diaphragm member 140 and the second diaphragm member 150 was performed in the same manner as in Experimental Example 2. Subsequently, signal comparisons were made before and after agitation.

FIG. 8 is a graph showing the results of detection of the luminescence intensity of each sample before, during and after stirring. As a result, it was confirmed that the signal was enhanced by the stirring. In FIG. 8, in the graphs of the 10000 pg / mL sample and the 1000 pg / mL sample, signals enhanced by stirring are indicated by double arrows.

DESCRIPTION OF

SYMBOLS

100, 500 ... Fluid device, 110, 110a, 110b, 110c, 110d ... Flow path, 115, 115a, 115b ... area |

region

120, 130, 510, 520, 530 ... Division valve, 140, 150 ... Diaphragm member, 160 ... Magnetic stand, 170: detection unit, 200, 300: diaphragm valve, 210: first substrate, 211: convex portion, 220: second substrate, 230: elastic member, 231: anchor portion, 240: through hole, L , L1, L2: fluid, M: magnetic particles, LB: washing buffer, 2nd Ab: secondary antibody, S: substrate reaction solution.

Claims

With the flow path,
A first compartment valve and a second compartment valve disposed in the flow path defining a predetermined area of the flow path by closing;
A first diaphragm member and a second diaphragm member disposed between the first partition valve and the second partition valve and forming at least a part of the flow channel wall of the flow channel;
A fluidic device capable of deforming at least one of the first diaphragm member and the second diaphragm member with the first partition valve and the second partition valve closed.
When the first diaphragm member or the second diaphragm member is deformed, deformation of the second diaphragm member or the first diaphragm member substantially changes the volume of the defined area. The fluidic device according to claim 1, which does not.
The said 1st diaphragm member and the said 2nd diaphragm member are deformable to bidirectional | two-way with the direction which goes to the axis | shaft of the said flow path, and the direction away from the axis | shaft of the said flow path. Fluid device.
The first diaphragm member deforms in a direction toward the axis of the flow path, and the second diaphragm member deforms in a direction away from the axis of the flow path, or the second diaphragm member moves in the axis of the flow path The fluid device according to claim 2 or 3, wherein the fluid device is deformed in the direction to move the first diaphragm member away from the axis of the flow passage.
A fluid device according to any of the preceding claims, wherein the first diaphragm member and the second diaphragm member are formed of an elastomeric material.
The fluid device according to any one of claims 2 to 5, wherein the amount of deformation of the first diaphragm member and the amount of deformation of the second diaphragm member are substantially the same.
The fluid device according to any one of claims 1 to 6, wherein the volume of the defined area is 10 μL or less.
The fluid device according to any one of claims 1 to 7, further comprising a detection unit between the first diaphragm member and the second diaphragm member in the defined area of the flow passage.
A first substrate and a second substrate bonded at a bonding surface,
At least one of the first substrate and the second substrate has a groove that forms a flow path by bonding the two substrates on the bonding surface,
The groove comprises two compartment valves,
The first substrate is
It has a first through hole and a second through hole disposed between the two partition valves and at a position facing the groove,
The opening on the groove side of the first through hole is closed by a first diaphragm member that is deformable in a direction toward the axis of the flow path,
A fluid device, wherein the opening on the groove side of the second through hole is closed by a second diaphragm member that is deformable in a direction away from the axis of the flow passage.
A method of stirring a fluid, wherein
Introducing the fluid into the flow path of the fluidic device according to any one of claims 1 to 9;
Closing the first compartment valve and the second compartment valve;
Deforming the first diaphragm member in a direction toward the axis of the flow passage, and repeating the process of eliminating the deformation of the first diaphragm member;
How to provide.
The method further includes the step of repeating the step of deforming the second diaphragm member in the direction toward the axis of the flow path and eliminating the deformation of the second diaphragm member.
The step of deforming the first diaphragm member to eliminate the deformation of the first diaphragm member, and the step of deforming the second diaphragm member to cancel the deformation of the second diaphragm member are alternated. The method according to claim 10, wherein:
The method further includes the step of repeating the step of deforming the first diaphragm member in a direction away from the axis of the flow path to eliminate the deformation of the first diaphragm member.
Deforming the first diaphragm member in a direction toward the axis of the flow channel to eliminate the deformation, and deforming the first diaphragm member in a direction away from the axis of the flow channel; The method according to claim 10, wherein the step of repeating the elimination of deformation of the diaphragm member is performed alternately.
The method further includes the step of deforming the second diaphragm member in a direction away from the axis of the flow passage, and repeating the process of removing the deformation of the first diaphragm member.
Deforming the first diaphragm member in a direction toward the axis of the flow channel to eliminate the deformation, and deforming the second diaphragm member in a direction away from the axis of the flow channel; The method according to any one of claims 10 to 12, wherein the steps of repeatedly canceling the deformation of the diaphragm member are performed alternately.
The method according to any one of claims 10 to 13, wherein the fluid comprises magnetic particles.