JPWO2019146102A1 - Fluid devices and their use - Google Patents

Abstract

A flow path, a first partition valve and a second partition valve that are arranged in the flow path and define a predetermined area of the flow path by closing, and the first partition valve and the second partition. A first diaphragm member and a second diaphragm member arranged between the valve and forming at least a part of the flow path wall of the flow path are provided, and the first compartment valve and the second compartment are provided. A fluid device capable of deforming at least one of the first diaphragm member and the second diaphragm member with the valve closed.

Description

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

In recent years, attention has been focused on the development of μ-TAS (Micro-Total Analysis Systems) aiming at speeding up, increasing efficiency, integration, and ultra-miniaturization of testing equipment in the field of in vitro diagnostics and biological research. Active research is underway worldwide.

μ-TAS is superior to conventional inspection equipment in that it can be measured and analyzed with a small amount of sample, that it can be carried, and that it can be disposable at low cost. In particular, it is attracting attention as a highly useful method when an expensive reagent is used or when a small amount of a large number of samples are tested.

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

Japanese Unexamined Patent Publication No. 2014-77810

The fluid device according to the embodiment includes a flow path, a first partition valve and a second partition valve arranged in the flow path, which define a predetermined area of the flow path by closing, and the first partition valve. The first diaphragm member and the second diaphragm member which are arranged between the partition valve and the second partition valve and form at least a part of the flow path wall of the flow path are provided. With the compartment valve and the second compartment valve closed, at least one of the first diaphragm member and the second diaphragm member can be deformed.

The fluid device according to one embodiment includes a first substrate and a second substrate to be joined at a joining surface, and at least one of the first substrate or the second substrate is flown by joining both substrates. A first substrate having a groove forming a path on the joint surface, the groove comprising two compartment valves, the first substrate being located between the two compartment valves and facing the groove. The opening on the groove side of the first through hole, which has a through hole and a second through hole, is closed by a first diaphragm member that is deformable in the direction toward the axis of the flow path, and the second through hole is formed. The groove-side opening of the through hole is closed by a second diaphragm member that is deformable in a direction away from the axis of the flow path.

The method for stirring the fluid according to the embodiment includes a step of introducing the fluid into the flow path of the fluid device, a step of closing the first partition valve and the second partition valve, and the first step. The diaphragm member is deformed in a direction toward the axis of the flow path, and the step of repeatedly eliminating the deformation of the first diaphragm member is provided.

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

Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings in some cases. In the drawings, the same or corresponding parts are designated by the same or corresponding reference numerals, and duplicate description will be omitted. The dimensional ratio in each figure is exaggerated for explanation and does not necessarily match the actual dimensional ratio.

[Fluid device]
(First Embodiment)
In one embodiment, the present invention comprises a flow path, a first partition valve and a second partition valve arranged in the flow path that define a predetermined region of the flow path by closing, and the first partition valve. The first diaphragm member and the second diaphragm member which are arranged between the partition valve and the second partition valve and form at least a part of the flow path wall of the flow path are provided. Provided is a fluid device capable of deforming at least one of the first diaphragm member and the second diaphragm member with the partition valve and the second partition valve closed.

In the fluid device, by closing the first partition valve and the second partition valve, a region including the first diaphragm member and the second diaphragm member of the flow path is defined, and the first partition valve is defined. By deforming the diaphragm member or the second diaphragm member, the fluid existing in the defined region is agitated.

The volume of the defined region, 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 a minute volume of fluid. In particular, in a fluid device such as the present invention, it has been difficult to agitate a minute volume of fluid on the flow path. Since the flow path diameter of the flow path into which a minute volume of fluid is introduced is narrowed by design, the fluid generally flows in a laminar flow in the flow path. Therefore, it is difficult to mix different liquids and liquids having different densities introduced into the flow path, and it takes a long time to stir so that the distribution becomes uniform.

On the other hand, as will be described later in the examples, the fluid device of the present embodiment can efficiently agitate a minute volume of fluid. The fluid to be agitated by the fluid device of the present embodiment may be a liquid or a gas. Further, when the liquid is agitated, the liquid may contain particles such as magnetic particles.

In the fluid device of the present embodiment, the flow path through which the fluid flows means a flow path for flowing the fluid, and includes both a state in which the fluid is flowing and a state in which the fluid is not flowing.

FIG. 1 is a top view and a cross-sectional view illustrating the structure of the fluid device 100 according to the 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 flow path 110 is provided with a first partition valve 120, a first diaphragm member 140, a second diaphragm member 150, and a second partition valve 130 in this order.

The cross section of the flow path 110 is, for example, substantially rectangular, circular, or elliptical. 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 in the flow path 110. By closing the first partition valve 120 and the second partition valve 130, the region 115 including the first diaphragm member 140 and the second diaphragm member 150 of the flow path 110 is defined, and the first diaphragm member 140 or the first diaphragm member 140 or Deformation of the second diaphragm member 150 agitates the fluid L present in the defined region 115.

In the example of FIG. 1, the first compartment valve 120 and the second compartment valve 130 are closed and the region 115 is defined. In the present specification, "definition" means to distinguish the target region of the flow path from other regions of the flow path, and can be referred to as "blocking", "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 a direction toward the axis of the flow path 110, a thickness direction of the second substrate 220 described later, and the like.

Changes in the volume of the defined region 115 caused by the deformation of the first diaphragm member 140 or the second diaphragm member 150, respectively, resulting in the deformation of the second diaphragm member 150 or the first diaphragm member 140, respectively. Is absorbed.

More specifically, when the first diaphragm member 140 is deformed in the direction toward the axis of the flow path 110 so as to protrude, the displaced fluid L directs the second diaphragm member 150 to the outside of the flow path 110. Extrude 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 move through the axis of the flow path 110. Deform in the direction away from. As a result, the volume of the region 115 is maintained at substantially the same volume as that before the deformation of the first diaphragm member 140. In this way, 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 and the volume of the region 115 is substantially changed. Not doing so is said to absorb changes in volume.

Similarly, when the second diaphragm member 150 is deformed so as to project in the direction toward the axis of the flow path 110, the displaced fluid L pushes the first diaphragm member 140 toward the outside of the flow path and is deformed. Let me. 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 move through the axis of the flow path 110. Deform in the direction away from. The volume of the region 115 is maintained at substantially the same volume as before the deformation of the second diaphragm member 150.

The fluid L existing in the region 115 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. It may take some time before the influence of the change in the volume of the region 115 is exerted. Therefore, a time difference may occur 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. ..

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

In the example of FIG. 1, the first diaphragm member 140 is deformed in the direction toward the axis of the flow path 110, and the second diaphragm member 150 is deformed in the direction away from the axis of the flow path 110. Then, the change in the 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 before the deformation of the first diaphragm member 140. Is maintained at the same volume as.

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, if the shape, size, material, 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 will be. It tends to be virtually the same. Here, substantially the same means that they are the same, or that they are substantially the same, and that there is a difference in the degree of error that is difficult to avoid in the manufacture of the fluid device.

In the fluid device of this embodiment, the volume of the defined region 115 and the volume of the agitated fluid L may be equal. When the defined region 115 is completely filled with the fluid L, the volume of the defined region 115 and the volume of the agitated fluid L are equal.

As will be 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. Further, when the first diaphragm member 140 or the second diaphragm member 150 is deformed while the region 115 is defined, the fluid L existing in the defined region 115 is deviated from the first diaphragm member 140. It moves in the direction toward the second diaphragm member 150 or in the direction from the second diaphragm member 150 toward the first diaphragm member 140 and is stirred.

《Partition 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 the region 115 can be defined, but may be, for example, a diaphragm valve. The distance between the first compartment valve 120 and the second compartment 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 arranged in the flow path 110 and form a part of the flow path 110. The fluid flowing through the flow path 110 comes into contact with the first partition valve 120 and the second partition valve 130, and the flow of the fluid is changed by the opening / 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 that they can be opened and closed in the flow path 110. Unlike the diaphragm member described later, the surface opposite to the flow path may be in contact with the substrate so as not to be deformed in the direction away from the axis of the flow path, and the valve is controlled to be deformed only in one direction. It may have been.

2 (a) and 2 (b) are cross-sectional views illustrating an example of the structure of the diaphragm valve. FIG. 2A shows an open state of the diaphragm valve 200, and FIG. 2B shows a closed state of the diaphragm valve 200. As shown in FIGS. 2A and 2B, the diaphragm valve 200 includes a first substrate 210, an elastic member 230 made of an elastomer material, and a second substrate 220. The second substrate 220 and the elastic member 230 are adhered to each other 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. Further, a through hole 240 is provided in a 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 valve control 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 the deformed elastic member A part of 230 is in close contact with the first substrate 210. This state is the closed state of the diaphragm valve 200. As a result, the flow of the fluid L inside the flow path 110 is blocked.

In the example of FIG. 2B, the diaphragm valve 200 is deformed in a direction perpendicular to the axial direction of the flow path 110 (the direction in which the fluid L flows) (the direction toward the axis of the flow path 110). Further, 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 opening and closing of the diaphragm valve 200 may be controlled by a fluid. The fluid valve control, N ₂ gas, a gas such as air, water, liquid such as oil and the like. The fluid for valve control can be supplied, for example, by a tube connected to the through hole 240 or the like. Alternatively, the opening and closing of the valve may be controlled by a mechanical force or an electromagnetic force.

The elastomer material for 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 a pressure change inside the through hole 240. For example, polydimethylsiloxane (PDMS). ), Silicone-based elastomers such as polymethylphenylsiloxane and polydiphenylsiloxane.

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

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

As shown in FIGS. 3A and 3B, in the diaphragm valve 300, the elastic member 230 is not arranged on the entire surface of the second substrate 220 as compared with the diaphragm valve 200 described above, and the diaphragm valve 300 has a diaphragm valve 300. The main difference is that it is locally located only around it.

Since the elastic member 230 of the diaphragm valve 300 includes the anchor portion 231 so that the elastic member 230 can be damaged 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 a fluid for valve control 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 the deformed elastic member A part of 230 is in close contact with the first substrate 210. This state is the closed state of the diaphragm valve 300. As a result, the flow of the fluid L inside the flow path 110 is blocked.

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

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

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

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 are used. By closing, the region 115 including the first diaphragm member 140 and the second diaphragm member 150 of the flow path 110 can be defined.

The first partition valve 120 and the second partition valve 130 may be a normally closed valve or a normally open valve. A normally closed valve is a valve that is in a closed state in a steady state and is opened by operating the valve. A normally open valve is a valve that is in an open state 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, the first partition valve 120 and the second partition valve 130 are in the closed state in the steady state, and the valves are operated. Releases the region 115 defined by the first compartment valve 120 and the second compartment valve 130.

Further, when the first partition valve 120 and the second partition valve 130 are normally open valves, the first partition valve 120 and the second partition valve 130 are in the open state in the steady state, and the valves are operated. By doing so, it becomes a closed state, and a part of the region 115 of the flow path 110 becomes a defined region.

<< 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 elastomer material. As the elastomer material, the same elastic member as described above can be used for the elastic member of the diaphragm valve. The opening and closing of the first diaphragm member 140 and the second diaphragm member 150 may be controlled by a fluid. The fluid valve control, N ₂ gas, a gas such as air, water, liquid such as oil and the like. Alternatively, the opening and closing of the valve may be controlled by a mechanical force or an electromagnetic force.

The first diaphragm member 140 and the second diaphragm member 150 are arranged in the flow path 110 and form a part of the flow path wall of the flow path 110. The fluid flowing through the flow path 110 comes into contact with the first diaphragm member 140 and the second diaphragm member 150, and the fluid flow changes due to the deformation of the first diaphragm member 140 and the second diaphragm member 150.

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 moves due to 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 path 110, and the second diaphragm member 150 can be deformed in the direction away from the axis of the flow path 110. Alternatively, for example, the second diaphragm member 150 can be deformed in the direction toward the axis of the flow path 110, and the first diaphragm member 140 can be deformed in the direction away from the axis of the flow path 110. Alternatively, both the first diaphragm member 140 and the second diaphragm member 150 can be deformed in both directions toward the axis of the flow path 110 and away from the axis of the flow path 110. This point is different from the above-mentioned point that the first partition valve 120 and the second partition valve 130 need to 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 be deformed to block the flow path 110. 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 being deformed, the first diaphragm member 140 and the second diaphragm member 150 are the same as those described above. 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 fluid device of the present embodiment, the region 115 may be a reaction section for performing an enzyme reaction, an antigen-antibody reaction, a nucleic acid hybridization reaction, or the like. As a result of stirring the fluid L existing in the region 115, the above reaction is uniformly performed, and effects such as improvement of reproducibility and enhancement of the detection signal can be obtained.

The fluid device of the present embodiment may further include a detection unit for detecting an enzyme reaction, an antigen-antibody reaction, a nucleic acid hybridization reaction, and the like. The detection unit exists 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 inside the region 115.

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

When the detection unit is present inside the region 115, the detection can be performed in real time even during stirring, that is, during execution of an enzyme reaction, an antigen-antibody reaction, a nucleic acid hybridization reaction, or the like. 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. Examples of the detector for optically detecting include, but are not limited to, a spectrophotometer, a fluorescence photometer, and the like.

《Fluid device》
The fluid device of the present embodiment includes a first substrate and a second substrate to be joined at a joining surface, and at least one of the first substrate or the second substrate is a flow path by joining both substrates. The groove comprises two compartment valves, the first substrate is located between the two compartment valves and at a position facing the groove. The opening on the groove side of the first through hole, which has a hole and a second through hole, is closed by a first diaphragm member that is deformable in the direction toward the axis of the flow path, and the second through. The groove-side opening of the hole is closed by a second diaphragm member that is deformable in a direction away from the axis of the flow path.

The fluid device of this 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 fluid device of this embodiment may be part of a fluid 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 laminated structure. A groove serving as a flow path 110 is formed on the joint surface of the two substrates. One substrate has at least two through holes, and a diaphragm member is provided on the joint surface side of each through hole. These diaphragm members become a first diaphragm member 140 and a second diaphragm member 150. As will be 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 each through hole is on the joint surface side. An elastic member may be provided. These elastic members become the first partition valve 120 and the second partition valve 130.

[How to stir the liquid]
(First Embodiment)
In one embodiment, the present invention is a method of stirring a fluid, the step of introducing the fluid into the flow path of the fluid device described above, and closing the first partition valve and the second partition valve. Provided is a method including a step of deforming the first diaphragm member in a direction toward the axis of the flow path and repeating the step of resolving the deformation of the first diaphragm member. By the method of this embodiment, a minute volume of fluid can be efficiently agitated.

The method of the present embodiment is a method of stirring a fluid, in which the step of introducing the fluid into the flow path of the fluid device described above and the first partition valve and the second partition valve are closed. The step of defining the region including the first diaphragm member and the second diaphragm member of the flow path and the process of deforming the first diaphragm member in the direction toward the axis of the flow path, and as a result, the definition. A step in which the fluid existing in the region moves in the direction from the first diaphragm member toward the second diaphragm member, and the second diaphragm member is deformed in a direction away from the axis of the flow path. , The deformation of the first diaphragm member is eliminated, and as a result, the fluid existing in the region moves in the direction from the second diaphragm member toward the first diaphragm member, and the second diaphragm member. Stirring by moving the fluid existing in the defined region from the first diaphragm member toward the second diaphragm member or in the opposite direction, comprising a step of eliminating deformation of the member. It may be the method to be done.

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 above-mentioned fluid device. As shown in FIG. 4A, first, the fluid L is introduced into the flow path 110 of the fluid device 100. The method of introducing the fluid L into the flow path 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 path 110.

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

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 a fluid similar to that described above as a fluid for valve control in the direction of arrow 140D1. Alternatively, the diaphragm member 140 may be deformed by a mechanical force or an electromagnetic force.

As a result, the fluid L existing in the defined region 115 moves in the direction from the first diaphragm member 140 toward the second diaphragm member 150 (direction indicated by the arrow LD1), and the second diaphragm member 150 flows. It is deformed in the direction away from the axis of the road 110 (the direction indicated by the arrow 150D1).

The change in the 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 the same as that before the deformation of the first diaphragm member 140. Maintained in the same volume.

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

As a result, the fluid L existing 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. To.

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

It is preferable that the step of deforming the first diaphragm member 140 in the direction toward the axis of the flow path 110 and the step of eliminating the deformation of the first diaphragm member 140 are repeated a plurality of times.

By the above steps, the fluid L existing in the region 115 moves from the first diaphragm member 140 toward the second diaphragm member 150 (direction indicated by the arrow LD1) or in the opposite direction (direction indicated by the arrow LD2). It is stirred by this.

By deforming and returning the first diaphragm member 140 in this way, the fluid L existing in the region 115 can be agitated.

(Second Embodiment)
The fluid agitation method according to the above-described embodiment further includes a step of repeatedly deforming the second diaphragm member in a 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 and eliminating the deformation of the first diaphragm member and the step of deforming the second diaphragm member and eliminating the deformation of the second diaphragm member are alternated. It may be done in.

For example, the second diaphragm member 150 is deformed in the direction toward the axis of the flow path 110, and as a result, the fluid L existing in the defined region 115 is directed from the second diaphragm member 150 toward the first diaphragm member 140. While moving in the direction (direction indicated by the arrow LD2), the step of deforming the first diaphragm member 140 in the direction away from the axis of the flow path 110 and the deformation of the second diaphragm member 150 are eliminated, and as a result, the region A step in which the fluid L existing in the 115 moves in the direction from the first diaphragm member 140 toward the second diaphragm member 150 (direction indicated by the arrow LD1) and the deformation of the first diaphragm member 140 is eliminated. May be further provided.

In the fluid agitation method according to the first embodiment, only the first diaphragm member 140 is deformed, whereas in the fluid agitation method according to the second embodiment, not only the first diaphragm member 140 but also the first diaphragm member 140 is deformed. The second diaphragm member 150 is also mainly different 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. 4 (a) to 4 (d) 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 in which the fluid L existing in the region 115 goes from the second diaphragm member 150 to the first diaphragm member 140. While moving in the direction (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 existing in the defined 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 first diaphragm member 140 flows. It is deformed in a direction away from the axis of the road 110 (direction indicated by arrow 140D2).

The change in the volume of the region 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 region 115 is the same as that before the deformation of the second diaphragm member 150. Maintained in the same volume.

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 supplied in the direction of the arrow 150D2 in FIG. 4 (e).

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

The change in the volume of the region 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 region 115 is the volume of the second diaphragm member 150. It is maintained at the same volume as before the deformation of.

In this way, the fluid L existing in the region 115 can be agitated by deforming the first diaphragm member 140 and the second diaphragm member 150 in the directions toward the axes of the flow paths 110 and returning them to their original positions. Can be done.

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

(Third Embodiment)
The fluid agitation method according to the first embodiment and the second embodiment described above is to deform the first diaphragm member in a direction away from the axis of the flow path to eliminate the deformation of the first diaphragm member. A step of deforming the first diaphragm member in a direction toward the axis of the flow path to eliminate the deformation and a step of deforming the first diaphragm member in a direction away from the axis of the flow path are further provided. The step of repeatedly deforming and eliminating the deformation of the first diaphragm member may be alternately performed.

For example, the fluid stirring method according to the first embodiment and the second embodiment described above deforms the first diaphragm member 140 in a direction away from the axis of the flow path 110 (direction indicated by the arrow 140D2), and as a result, the result is The fluid L existing in the defined region 115 moves in the direction from the second diaphragm member 150 toward the first diaphragm member 140 (direction indicated by the arrow LD2), and the second diaphragm member 150 moves in the flow path 110. The step of deforming in the direction toward the axis (direction indicated by the arrow 150D2) and the deformation of the first diaphragm member 140 are eliminated, and as a result, the fluid L existing in the region 115 is transferred from the first diaphragm member 140 to the second diaphragm member 140. It may further include a step of moving in the direction toward the diaphragm member 150 (direction indicated by the arrow 150D1) and eliminating the deformation of the second diaphragm member 150.

The fluid agitation method according to the third embodiment is compared with the fluid agitation method according to the first embodiment and the second embodiment, in the direction in which the first diaphragm member 140 is directed toward the axis of the flow path 110 and the flow path. It differs mainly 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 path 110.

Here, deforming means actively deforming the diaphragm member, and does not include passively deforming the diaphragm member in order to absorb the change 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 embodiment and the second embodiment for the steps described above with reference to FIGS. 4 (a) to 4 (d).

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

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 a mechanical force or an electromagnetic force.

As a result, the fluid L existing in the defined 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 second diaphragm member 150 flows. It deforms in the direction toward the axis of the road 110.

In this step, 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 step described with reference to FIG. 4 (e) for the second embodiment. In the second embodiment, the second diaphragm member 150 is actively deformed, whereas in the third embodiment, the first diaphragm member 140 is actively deformed.

The change in the 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 the same as that before the deformation of the first diaphragm member 140. Maintained in the same volume. 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 deformation of the first diaphragm member 140 can be eliminated, for example, by stopping the suction in the direction of the arrow 140D2 described above in FIG. 4 (e).

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

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

In this way, the fluid L existing in the region 115 is also agitated by deforming the first diaphragm member 140 in the direction toward the axis of the flow path 110 and the direction away from the axis of the flow path 110 and returning the first diaphragm member 140 to its original position. 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 path, and the first diaphragm member Further comprising a step of repeating the process of eliminating the deformation, the step of deforming the first diaphragm member in the direction toward the axis of the flow path to eliminate the deformation, and the step of removing the deformation of the second diaphragm member of the flow path. The step of repeatedly deforming the second diaphragm member in the direction away from the shaft and eliminating the deformation of the second diaphragm member may be alternately performed.

For example, 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), and as a result, the fluid L existing in the defined region 115 is the first from the first diaphragm member 140. A step of moving in the direction toward the diaphragm member 150 of 2 (the direction indicated by the arrow LD1) and deforming the first diaphragm member 140 in the direction toward the axis of the flow path 110 (the direction indicated by the arrow 140D1), and a second. As a result, the fluid L existing in the region 115 moves in the direction from the second diaphragm member 150 toward the first diaphragm member 140 (direction indicated by the arrow LD2), and at the same time, the first diaphragm member 150 is deformed. It may further include a step of eliminating the deformation of the diaphragm member 140 of 1.

The fluid agitation method according to the fourth embodiment has the second diaphragm member 150 on the axis of the flow path 110 as compared with the fluid agitation method according to the first embodiment, the second embodiment and the third embodiment. It differs mainly in that it deforms in both directions in the direction toward which the flow path 110 is directed and in the direction away from the axis of the flow path 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.

Here, deforming means actively deforming the diaphragm member, and does not include passively deforming the diaphragm member in order to absorb the change in the volume of the region 115.

The fluid agitation method according to the fourth embodiment will be described with reference to FIGS. 4 (a) to 4 (f). The method for stirring the fluid according to the fourth embodiment is the method for stirring the fluid according to the first embodiment, the second embodiment and the third embodiment for the above-described steps with reference to FIGS. 4 (a) to 4 (b). Is similar to.

As described above with reference to FIG. 4B, the first compartment valve 120 and the second compartment valve 130 are closed to include the first diaphragm member 140 and the second diaphragm member 150 of the flow path 110. When the region 115 is defined, the flow of the fluid L existing inside the region 115 is stopped, and the region 115 becomes stationary.

Subsequently, as shown in FIG. 4C, the second diaphragm member 150 is deformed in a direction away from the axis of the flow path 110 (direction indicated by 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 a mechanical force or an electromagnetic force.

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

In this step, 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 FIGS. 4 (c) for the first embodiment, the second embodiment, and the third embodiment. It is the same as that in the process. In the first embodiment, the second embodiment and the third embodiment, the second diaphragm member 150 is passively deformed, whereas in the fourth embodiment, the second diaphragm member 150 is actively deformed. The point is different.

The change in the volume of the region 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 region 115 is the same as that before the deformation of the second diaphragm member 150. Maintained in the same volume. 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 in FIG. 4C.

As a result, the fluid L existing 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. To.

The change in the volume of the region 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 region 115 is the volume of the second diaphragm member 150. It is maintained at the same volume as before the deformation of.

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 a fluid similar to that described above as the fluid for valve control in the direction of the arrow 150D2. Alternatively, the diaphragm member 150 may be deformed by a mechanical force or an electromagnetic force.

As a result, the fluid L existing in the defined 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 first diaphragm member 140 flows. It is deformed in a direction away from the axis of the road 110 (direction indicated by arrow 140D2).

In this step, 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 step described with reference to FIG. 4 (e) for 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.

The change in the volume of the region 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 region 115 is the same as that before the deformation of the second diaphragm member 150. Maintained in the same volume. Here, the deformation of the first diaphragm member 140 is passive.

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 supplied in the direction of the arrow 150D2 in FIG. 4 (e).

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

The change in the volume of the region 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 region 115 is the volume of the second diaphragm member 150. It is maintained at the same volume as before the deformation of.

In this way, the fluid L existing in the region 115 is also agitated by deforming and returning the second diaphragm member 150 in the direction toward the axis of the flow path 110 and the direction away from the axis of the flow path 110. be able to.

The fluid agitation methods according to the first embodiment, the second embodiment, the third embodiment and the fourth embodiment described above may be carried out in combination with each other.

For example, active deformation of the first diaphragm member 140 toward the axis of the flow path 110 and away from the axis of the flow path 110, and the direction of the second diaphragm member 150 toward the axis of the flow path 110 and Active deformation in the direction away from the axis of the flow path 110 may be carried out 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 regarded as one cycle, and the first embodiment, the second embodiment, the third embodiment, and the fourth embodiment described above are defined for each cycle. It may be carried out while switching the method of stirring the fluid according to the embodiment.

In the method for stirring the fluid according to the first embodiment, the second embodiment, the third embodiment, and the fourth embodiment described above, the fluid L to be agitated may be a liquid or a gas. , It may be a powder or granular material. For example, the fluid L is a plurality of types of liquids, and a plurality of types of liquids may be mixed by the stirring method of the present invention. Further, the fluid L may be a liquid or a gas having a non-uniform concentration, temperature, and distribution state of components, and the distribution state may be made uniform by the stirring method of the present invention. Further, the fluid L is a liquid having a non-uniform distribution state of a dispersion system such as an emulsion, and the distribution state may be made uniform or finer by the stirring method of the present invention. Further, 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 will be described later in the examples, the fluid L can be efficiently agitated even when the fluid L contains magnetic particles. Further, since the magnetic particles can be accumulated by using a magnetic force, for example, the magnetic particles can be accumulated inside the region 115. Further, the fluid L containing the magnetic particles can be efficiently agitated by the method described above. Further, for example, a magnet (not shown) may be brought close to the region 115 to exert a magnetic force to accumulate magnetic particles in the region 115 in advance. Then, the fluid L is introduced into the region 115, the first partition valve 120 and the second partition valve 130 are closed to define the region 115, and then the magnet is released to release the magnetic particles in the region 115. The magnetic particles may be stirred in the fluid L by the stirring method of.

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

(Fifth Embodiment)
The fluid agitation method according to the fifth embodiment is a method of quantifying and mixing two types of fluids. The method for stirring the fluid 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 the embodiment. The fluid device 500 includes flow paths 110a, 110b, 110c, 110d for flowing a fluid, a first partition valve 120, a second partition valve 130, a third partition valve 510, and a fourth partition valve. It includes a 520, a fifth partition valve 530, a first diaphragm member 140, and a second diaphragm member 150.

Hereinafter, a procedure for 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 flowed 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. As a result, in the flow path 110a, the 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 existing in the region 115a is quantified. ..

On the other hand, as shown in FIGS. 5A or 5B, 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 flowed 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. As a result, in the flow path 110c, the 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 existing in the region 115b is quantified. ..

Subsequently, as shown in FIG. 5D, the third partition valve 510 is opened. As a result, the flow path 110a and the flow path 110b 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 contains quantified fluid L1 and fluid L2, respectively.

Subsequently, by deforming the first diaphragm member 140 or the second diaphragm member 150, the fluid L1 and the fluid L2 existing in the defined region 115 are agitated and mixed. Here, the method of deforming the first diaphragm member 140 and the second diaphragm member 150 is the same as the method of stirring the fluid 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 area 115. In this case, the state of the fluid inside the region 115 can be detected and measured by using the detector during or after stirring the fluid L1 and the fluid L2.

[Analysis method]
In one embodiment, the present invention provides an analysis method comprising a step of stirring a fluid by the method described above and a step of analyzing the stirred fluid.

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

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. An antibody against the protein to be measured is bound to the surface of the magnetic particles M.

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

Subsequently, as shown in FIG. 6B, the cleaning buffer LB may be flowed through the flow path 110 of the fluid device 100 to clean the magnetic particles M.

Subsequently, as shown in FIG. 6B, a sample containing the sample T is flowed through the flow path 110 of the fluid device 100. Sample T is a molecule to be detected to which an antibody bound to the surface of magnetic particles M specifically binds.

Subsequently, as shown in FIG. 6C, 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 sample containing the sample T and the magnetic particles M are separated inside the region 115. Further, here, the diaphragm members 140 and 150 of the fluid device 100 are operated, and the fluid L (the liquid containing the sample T and the magnetic particles M) inside the region 115 is agitated as described above. As a result, the fluid L inside the region 115 becomes uniform, and the antibody and the sample T arranged on the surface of the magnetic particles M react with each other with good reproducibility and efficiency.

Subsequently, as shown in FIG. 6B, the fluid device 100 is set on the magnetic stand 160 to accumulate the magnetic particles M existing in the region 115. Further, as shown in FIG. 6B, a cleaning buffer LB is passed through the flow path 110 to clean the magnetic particles M.

Subsequently, as shown in FIG. 6B, a buffer containing the secondary antibody (2ndAb) is flowed through the flow path 110 of the fluid device 100. The secondary antibody is a labeled antibody that specifically binds to sample T.

Subsequently, as shown in FIG. 6C, 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 separated inside the region 115. Further, here, the diaphragm members 140 and 150 of the fluid device 100 are operated to stir the fluid L (the 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 particles M and the secondary antibody react with each other with good reproducibility and efficiency.

Subsequently, as shown in FIG. 6B, the fluid device 100 is set on the magnetic stand 160 to accumulate the magnetic particles M existing in the region 115. Further, as shown in FIG. 6B, a cleaning buffer LB is passed through the flow path 110 to clean the magnetic particles M.

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

Subsequently, as shown in FIG. 6D, 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 separated inside the region 115. Further, here, the diaphragm members 140 and 150 of the fluid device 100 are operated, and as described above, the fluid L (the liquid containing the substrate reaction solution S and the magnetic particles M to which the secondary antibody is bound) inside the region 115 is applied. Stir. As a result, the fluid L inside the region 115 becomes uniform, and the secondary antibody bound to the surface of the magnetic particles M and the substrate reaction solution S react with each other with good reproducibility and efficiency.

As a result, a signal is generated. The signal is, for example, fluorescence. Then, the generated signal is detected. The signal may be detected, for example, by using a detector 170 installed adjacent to the region 115, or a detector (not shown) installed outside the fluid device, as shown in FIG. 6 (d). May be detected using.

When the detector is installed outside the fluid device, the reaction solution after the reaction inside the region 115 shown in FIG. 6 (d) is sent to a position that can be detected by the detector and then moved. It is good to detect the signal.

Although the specific example of carrying out the antigen-antibody reaction has been described above by the analysis method of the present embodiment, an enzyme reaction, a nucleic acid hybridization reaction and the like can also be carried out by the same method. Further, in the above-mentioned 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 solution S were carried out using the fluid device 100. The analysis method is not limited to this, and for example, only the reaction between the secondary antibody and the substrate reaction solution S may be carried out using the fluid device 100, and the other reactions may be carried out by another fluid device or the like.

Next, the present invention will be described in more detail with reference to Examples, but the present invention is not limited to the following Examples.

[Experimental Example 1]
(Manufacturing of fluid devices)
A fluid device whose schematic diagram is shown in FIGS. 1 (a) and 1 (b) was manufactured. In the manufactured fluid device (hereinafter referred to as "device A"), the cross section of the flow path 110 is substantially rectangular, the width of the flow path 110 is 0.5 to 1.0 mm, and the height is 0.2 to 0.2. 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 the 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. It was.

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 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 agitated using the device A manufactured in Experimental Example 1. First, Cy5 (Dojin Kagaku Kenkyusho), which is a fluorescent dye labeled with biotin, is immobilized on magnetic particles (JSR) having a diameter of 3 μm modified with streptavidin by an avidin-biotin bond. Magnetic particles were prepared. The prepared fluorescent magnetic particles are TBST (0.05% Tween 20-Tris Buffered Saline) buffer containing 0.1% BSA (bovine serum albumin) so as to have a final concentration of 0.01% (hereinafter, "0.1%". It was suspended in BSA-TBST) and used in the following experiments.

Subsequently, a neodymium magnet having a size of 3 mm × 3 mm × 5 mm 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 flow path 110. Then, 10 μL of 0.1% BSA-TBST was introduced to fill the flow path. FIG. 7A is a photomicrograph of the device A after introducing the fluorescent magnetic particles. The first partition valve 120 and the second partition valve 130 are closed and partitioned.

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

Here, of the first diaphragm member 140 and the second diaphragm member 150, the non-operating diaphragm member was passively deformed in a direction of absorbing the volume change of the region 115.

FIG. 7B is a photomicrograph of the device A after stirring. As a result, as shown in FIG. 7B, it was clarified that the magnetic particles were uniformly agitated in an extremely short time (about 1 second).

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

[Experimental Example 3]
(Detection of antigen-antibody reaction)
The antigen-antibody reaction was detected using the device A produced in Experimental Example 1. As the antigen, myocardial troponin I (cardial troponin I, cTnI) was used.

First, an anti-cTnI antibody (HyTest) was bound to a magnetic particle (JSR) having a surface modified with a carboxyl group and having a diameter of 3 μm via an 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 anti-cTnI antibody (HyTest) having an epitope different from that of the anti-cTnI antibody bound to the magnetic particles was labeled with alkaline phosphatase using an alkaline phosphatase labeling kit (Dojin Chemical Research Institute).

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

Subsequently, the magnetic particles were washed three times with 200 μL of 0.1% BSA-TBST while capturing the magnetic particles using 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 flow path 110 of the device A, and the magnetic particles were accumulated in the region 115.

Subsequently, 10 μL of CDP-star (Thermo Fisher Scientific), which is a chemiluminescent substrate for alkaline phosphatase, was introduced into the flow path 110 of the device A, and the first partition valve 120 and the second partition valve 130 were closed. The region 115 was determined, and luminescence due to the decomposition reaction of CDP-star was detected in real time. ΜPMT (Hamamatsu Photonics Co., Ltd.) was used as the detector. The detector was placed so that the sensor was adjacent to the area 115 of device A and the measurement was performed.

Three minutes after the start of the measurement, the first diaphragm member 140 and the second diaphragm member 150 were operated, and stirring was performed for 90 seconds. The operation of the first diaphragm member 140 and the second diaphragm member 150 was carried out in the same manner as in Experimental Example 2. Subsequently, the signals were compared before and after stirring.

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

100,500 ... Fluid device, 110, 110a, 110b, 110c, 110d ... Flow path, 115, 115a, 115b ... Region, 120, 130, 510, 520, 530 ... Partition 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 ... cleaning valve, 2ndAb ... secondary antibody, S ... substrate reaction solution.

Claims (14)

Channel and
A first partition valve and a second partition valve arranged in the flow path that define a predetermined area of the flow path by closing.
A first diaphragm member and a second diaphragm member arranged between the first partition valve and the second partition valve and forming at least a part of the flow path wall of the flow path are provided.
A fluid device capable of deforming at least one of the first diaphragm member and the second diaphragm member with the first compartment valve and the second compartment valve closed. When the first diaphragm member or the second diaphragm member is deformed, the volume of the defined region is substantially changed by the deformation of the second diaphragm member or the first diaphragm member. The fluid device according to claim 1. The first or second diaphragm member, wherein the first diaphragm member and the second diaphragm member are deformable in both directions in a direction toward the axis of the flow path and a direction away from the axis of the flow path, according to claim 1 or 2. Fluid device. The first diaphragm member is deformed in the direction toward the axis of the flow path, and the second diaphragm member is deformed in the direction away from the axis of the flow path, or the second diaphragm member is deformed in the direction away from the axis of the flow path. The fluid device according to claim 2 or 3, which deforms in a direction toward which the first diaphragm member deforms in a direction away from the axis of the flow path. The fluid device according to any one of claims 1 to 4, wherein the first diaphragm member and the second diaphragm member are formed of an elastomer 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 region is 10 μL or less. The fluid device according to any one of claims 1 to 7, wherein a detection unit is provided between the first diaphragm member and the second diaphragm member in the defined region of the flow path. A first substrate and a second substrate to be joined at a joining surface are provided.
At least one of the first substrate or the second substrate has a groove on the joint surface that forms a flow path by joining both substrates.
The groove comprises two compartment valves
The first substrate is
It has a first through hole and a second through hole that are located between the two compartment valves and at positions facing the groove.
The groove-side opening of the first through hole is closed with a first diaphragm member that is deformable in the direction toward the axis of the flow path.
A fluid device in which the groove-side opening 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 path. A method of stirring a fluid
A step of introducing the fluid into the flow path of the fluid device according to any one of claims 1 to 9.
The step of closing the first partition valve and the second partition valve, and
A step of repeatedly deforming the first diaphragm member in a direction toward the axis of the flow path and eliminating the deformation of the first diaphragm member, and
How to prepare. A step of repeatedly deforming the second diaphragm member in the direction toward the axis of the flow path and eliminating the deformation of the second diaphragm member is further provided.
The step of deforming the first diaphragm member and eliminating the deformation of the first diaphragm member and the step of deforming the second diaphragm member and eliminating the deformation of the second diaphragm member are alternated. 10. The method according to claim 10. Further comprising a step of repeatedly deforming the first diaphragm member in a direction away from the axis of the flow path and eliminating the deformation of the first diaphragm member.
The step of deforming the first diaphragm member in a direction toward the axis of the flow path to eliminate the deformation, and the step of deforming the first diaphragm member in a direction away from the axis of the flow path, the first The method according to claim 10 or 11, wherein the steps of repeatedly eliminating the deformation of the diaphragm member are alternately performed. Further comprising a step of repeatedly deforming the second diaphragm member in a direction away from the axis of the flow path and eliminating the deformation of the first diaphragm member.
The step of deforming the first diaphragm member in the direction toward the axis of the flow path to eliminate the deformation, and the step of deforming the second diaphragm member in the direction away from the axis of the flow path, and the second The method according to any one of claims 10 to 12, wherein the step of repeatedly eliminating the deformation of the diaphragm member is alternately performed. The method according to any one of claims 10 to 13, wherein the fluid contains magnetic particles.