WO2018235634A1

WO2018235634A1 - Fluid device and use thereof

Info

Publication number: WO2018235634A1
Application number: PCT/JP2018/022032
Authority: WO
Inventors: 慶治三井; 遼小林; 哲臣高崎
Original assignee: 株式会社ニコン
Priority date: 2017-06-21
Filing date: 2018-06-08
Publication date: 2018-12-27
Also published as: JPWO2018235634A1; JP7173004B2

Abstract

This fluid device includes: a first fluid introducing portion into which a first fluid containing a substance to be separated is introduced; a second fluid introducing portion into which a second fluid for recovering the substance to be separated is introduced; and a fluid adjusting portion which adjusts the flow of the first fluid and/or the flow of the second fluid; wherein at least one of the first fluid introducing portion and the second fluid introducing portion has a flow passage shape, and at least parts of the first fluid introducing portion and the second fluid introducing portion communicate with one another across a filter which selectively allows the passage of the substance to be separated, or a filter which selectively prevents the passage of a prescribed substance other than the substance to be separated.

Description

Fluid device and use thereof

The present invention relates to fluidic devices and their use. Priority is claimed on Japanese Patent Application No. 2017-121353, filed Jun. 21, 2017, the content of which is incorporated herein by reference.

In a clinical examination, blood cell components are separated from blood, and plasma and serum are collected. In general, blood cell components in blood are precipitated by centrifugation using a centrifuge, and the supernatant is often collected to obtain plasma or serum. However, with this method, separation of blood cell components takes time, and processing of a small amount of sample is difficult.

Also known is a method of separating blood cell components from blood through a filter. However, in this method, pressure is applied in the filtration direction of the filter, and the blood cell component is easily clogged in the filter, and hemolysis is likely to occur. In order to suppress clogging of the filter, it has been proposed to use an asymmetric porous membrane in which the pore size changes in the thickness direction (see, for example, Patent Document 1), but the structure becomes complicated and hemolysis is sufficiently It can not be prevented either.

There is a need for new devices that can efficiently separate blood cell components from small amounts of blood without hemolysis.

Japanese Patent Application Publication No. 11-505327

A fluidic device according to one embodiment includes a first fluid introducing part into which a first fluid containing a substance to be separated is introduced, and a second fluid introducing part into which a second fluid to collect the substance to be separated is introduced. And / or a fluid regulator that regulates the flow of the first fluid and / or the flow of the second fluid, and at least one of the first fluid inlet and the second fluid inlet has a flow path shape. And at least a portion of the first fluid inlet and the second fluid inlet selectively pass a substance to be separated, or a predetermined substance other than the substance to be separated. They are in communication with one another through non-permeable filters.

The method of separating a substance to be separated according to one embodiment is a method of separating a substance to be separated from a first fluid containing the substance to be separated using a fluid device, wherein the fluid device comprises A first fluid introducing portion to which a fluid is introduced, a second fluid introducing portion to which a second fluid for recovering the substance to be separated is introduced, a flow of the first fluid, and / or the second fluid And at least one of the first fluid introduction part and the second fluid introduction part has a flow path shape, and the first fluid introduction part and the second fluid introduction part At least a part of which is a filter that selectively transmits the substance to be separated or a fluidic device in communication with each other via a filter that does not selectively transmit substances other than the substance to be separated; The first fluid introducing portion Introducing the second fluid into the second fluid introduction part, and controlling the flow of the first fluid and / or the flow of the second fluid; And activating the first fluid and / or the second fluid, and recovering the fluid in the second fluid inlet.

A fluidic device according to one embodiment is a separation unit that separates a substance to be separated from a first fluid containing the substance to be separated, the first flow path for flowing the first fluid, and the substance to be separated A second flow path for flowing the fluid to be recovered, a pump unit for adjusting the flow of the first fluid, and / or a pump unit for adjusting the flow of the second fluid, wherein the first flow At least a part of the passage and the second flow path communicate with each other through a filter that selectively transmits the substance to be separated or a filter that does not selectively transmit a predetermined substance other than the substance to be separated Including the separation unit.

The method of separating a substance to be separated according to one embodiment is a method of separating a substance to be separated from a first fluid containing the substance to be separated using a fluid device, wherein the fluid device comprises the substance to be separated. A separation unit for separating a substance to be separated from a first fluid containing the first fluid, and a second flow path for flowing a second fluid for recovering the substance to be separated; And / or a pump portion that regulates the flow of the first fluid, and / or a pump portion that regulates the flow of the second fluid, and at least one of the first flow passage and the second flow passage. A part includes a separation part communicating with each other through a filter that selectively transmits the substance to be separated or a filter that does not selectively transmit other substances than the substance to be separated; Introducing the first fluid into the first flow channel; Introducing the second fluid into the passage, and operating the pump unit that regulates the flow of the first fluid and / or the pump unit that regulates the flow of the second fluid to operate the first fluid And / or displacing the second fluid, and recovering the second fluid.

FIG. 2 is a schematic view illustrating an embodiment of a fluidic device. It is a schematic diagram which shows the modification of a fluid device. (A) And (b) is sectional drawing explaining the structure of a diaphragm valve. (A)-(d) is a sectional view explaining operation of a pump provided with three diaphragm valves. (A) And (b) is sectional drawing explaining the structure of one Embodiment of a diaphragm valve. (A) is a perspective view explaining one embodiment of a separation part. (B) is an arrow sectional view in the b-b 'line of (a). It is a photograph of the isolation part manufactured by example 1 of an experiment. It is a photograph of the isolation | separation part which introduce | transduced simulated blood into the 1st flow path in Experimental example 2, and introduce | transduced the phosphate buffer (PBS) into the 2nd flow path. In Experimental example 2, it is a photograph of the isolation | separation part about 250 seconds after an experiment start. It is a graph which shows the result of example 2 of an experiment.

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
In the present embodiment, the fluidic device includes a first fluid introducing portion into which a first fluid containing a substance to be separated is introduced, and a second fluid introducing portion into which a second fluid to collect the substance to be separated is introduced. And / or a fluid regulator that regulates the flow of the first fluid and / or the flow of the second fluid, and at least one of the first fluid inlet and the second fluid inlet has a flow path shape. And at least a portion of the first fluid inlet and the second fluid inlet selectively pass a substance to be separated, or a predetermined substance other than the substance to be separated. They are in communication with one another through non-permeable filters.

The fluid device of this embodiment can also be said to have a separation part for separating the substance to be separated from the fluid. In this case, the separation unit includes a first flow path for flowing a first fluid containing a separation material, a second flow path for flowing a second fluid for recovering the separation material, and the first fluid And / or a pump unit for controlling the flow of the second fluid, wherein at least a portion of the first flow passage and the second flow passage are separated The filters are in communication with each other through filters that selectively transmit substances or filters that do not selectively transmit predetermined substances other than the substance to be separated.

FIG. 1 is a schematic view illustrating a fluid device of the present embodiment. As shown in FIG. 1, the fluid device includes a first fluid introducing unit 110, a second fluid introducing unit 120, and a fluid adjusting unit 130. The fluid device includes a first fluid introducing unit 110 and a second fluid introducing unit 120. At least a portion is in communication with each other via the filter 140.

FIG. 1 can also be said to be a schematic view showing an example of the separation part of the fluid device of the present embodiment. As shown in FIG. 1, the separation unit 100 includes a first flow passage (first fluid introduction unit) 110, a second flow passage (second fluid introduction unit) 120, and a pump unit (fluid adjustment unit) 130. And a part of the first channel 110 and the second channel 120 are in communication with each other through the filter 140.

The first flow path 110 is for flowing a fluid containing a substance to be separated. The second flow path 120 is for flowing a fluid for recovering the substance to be separated. That is, each of the first flow path (first fluid introduction portion) and the second flow path (second fluid introduction portion) may have a flow path shape. The pump unit 130 also regulates the flow of fluid in the first flow passage 110. In addition, the filter 140 is a filter that selectively transmits the substance to be separated or does not selectively transmit a predetermined substance other than the substance to be separated. The first fluid introduction portion and the second fluid introduction portion may have portions that are substantially parallel, and a filter may be provided on the parallel portions.

As shown in FIG. 1, an inflow channel 111 for introducing a fluid into the first channel 110 may be connected to the first channel 110. Further, a discharge flow path 112 for discharging the fluid in the first flow path 110 to the outside may be connected to the first flow path 110. In addition, the separation unit 100 may include a valve 113 for controlling the communication between the inflow channel 111 and the first channel 110 in an open state or a closed state. In addition, the separation unit 100 may include a valve 114 for controlling the communication between the discharge flow path 112 and the first flow path 110 in an open state or a closed state.

In addition, an inflow channel 121 for introducing a fluid into the second flow channel 120 may be connected to the second flow channel 120. Further, a discharge flow path 122 for discharging the fluid inside the second flow path 120 to the outside may be connected to the second flow path 120. In addition, the separation unit 100 may include a valve 123 for controlling the communication between the inflow channel 121 and the second channel 120 in an open state or a closed state. Further, the separation unit 100 may include a valve 124 for controlling the communication between the discharge flow path 122 and the second flow path 120 in an open state or a closed state.

Although any valve used for a fluid device can be used as

valve

113, 114, 123, 124, For example, the thing similar to a valve which constitutes pump part 130 mentioned below can be used.

Here, a method of separating the substance to be separated from the first fluid containing the substance to be separated using the separation unit 100 will be described by way of an example.

First, a first fluid containing a substance to be separated is introduced into the first flow path 110 of the separation unit 100. The substance to be separated is not particularly limited, but here, as an example, a case where plasma components are separated and collected as a substance to be separated from a blood sample will be described. That is, here, the first fluid containing the substance to be separated is a blood sample. The blood sample may be whole blood, or whole blood may have some components removed by pretreatment. Blood contains blood cell components such as red blood cells, white blood cells, and platelets in addition to plasma components, and for clinical examination, it may be necessary to recover plasma components from which red blood cells, white blood cells, and platelets have been removed. .

First, both the valve 113 and the valve 114 are opened. As a result, the inflow channel 111, the first channel 110, and the discharge channel 112 are in communication. Subsequently, a blood sample is introduced from the inflow channel 111. After the introduced blood sample fills the first flow path 110, the surplus is discharged from the discharge flow path 112. When the first channel 110 is filled with the blood sample, both the valve 113 and the valve 114 are closed. As a result, the first flow path 110 forms a closed circuit.

As will be described later, by forming a circuit in which the first flow path 110 is closed, it becomes possible to circulate the fluid in the first flow path 110 repeatedly, and separation of the substance to be separated is efficiently performed. Becomes easier.

Note that the first flow path 110 may not form a closed circuit. It is also possible to separate the substance to be separated by flowing the first fluid in one direction without looping the first flow path.

Subsequently, a fluid for recovering the substance to be separated is introduced into the second flow passage 120. In this example, for example, physiological saline may be used as a fluid for dissolving or dispersing and collecting plasma components. The order of introducing the fluid into the first channel 110 and the second channel 120 is not limited.

First, both the valve 123 and the valve 124 are opened. As a result, the inflow channel 121, the second flow channel 120, and the discharge channel 122 are in communication. Subsequently, physiological saline is introduced from the inflow channel 121. After the introduced saline fills the second flow path 120, the surplus is discharged from the discharge flow path 122. When the second flow path 120 is filled with saline, both the valve 123 and the valve 124 are closed. As a result, the second flow passage 120 forms a closed circuit.

As described later, by forming a circuit in which the second flow passage 120 is closed, it is possible to repeatedly circulate the fluid in the second flow passage 120, and the separation and concentration of the substance to be separated can be efficiently performed. It will be easy to do.

Note that the second flow path 120 may not form a closed circuit. It is also possible to recover the substance to be separated by flowing the second fluid in one direction without looping the second flow path.

Subsequently, the pump unit 130 is operated. As a result, a first fluid, ie, a blood sample, circulates inside the first channel 110. As a result, the blood cell component receives a force in the flow path direction, that is, in a direction parallel to the filter 140, and the force in the filtration direction of the filter is not easily received. Therefore, it is possible to suppress clogging of the filter by the blood cell component and breakage of the blood cell component to cause hemolysis.

On the other hand, the substance to be separated which can pass through the filter 140 moves from the first flow path 110 to the second flow path 120. As described later, when using a plurality of valves disposed in the first flow path 110 as the pump unit 130, the volume of the first flow path may be slightly reduced by operating the pump and closing the valve. Thus, the substance to be separated is appropriately pushed out to the second flow path, and the substance to be separated can be efficiently separated.

The filter 140 may selectively transmit only the substance to be separated, or may not selectively transmit a predetermined substance other than the substance to be separated. In the latter case, it is possible to use, as the filter 140, one that does not selectively permeate a predetermined substance that is not desired to be contained in the substance to be separated. In this case, in addition to the substance to be separated, other substances which may not be separated from the substance to be separated may pass through the filter 140. In the case of this example, as the filter 140, a filter that does not selectively permeate red blood cells, white blood cells and platelets but allows plasma components (substance to be separated) to pass is used. For example, a filter sheet having a pore diameter of about 1 μm can be used. Details of the filter will be described later.

Subsequently, the second fluid in the second channel 120, that is, the saline containing the plasma component is recovered.

First, both the valve 123 and the valve 124 are opened. As a result, the inflow channel 121, the second flow channel 120, and the discharge channel 122 are in communication. Subsequently, the saline solution in which the plasma component is dissolved or dispersed is discharged from the discharge channel 122 and collected. By the above operation, plasma components can be separated from the blood sample.

The substance to be separated which can be separated by the separation unit 100 of the present embodiment is not limited to the plasma component. For example, the liposomes in the solution can be fractionated according to size, or the beads can be separated from the liquid mixture containing the beads, or the fine particles (garbage), bacteria, and viruses are separated from the environment or gases such as exhalation. Can also be used to That is, as the substance to be separated, plasma components, liposomes, beads, other fine particles, microorganisms and the like can be mentioned.

In the present embodiment, the object is to recover a substance to be separated (plasma component). Therefore, after the components other than the substance to be separated sufficiently move to the second channel 120, the fluid in the first channel is recovered. In addition, after the substance to be separated has moved to the second flow path, the substance remaining in the first flow path may be recovered.

Thus, a new separation part can be provided by the present embodiment. According to the fluid device provided with the separation unit of the present embodiment, for example, clinical examination using a small amount of blood can be performed on a small device. In this case, the width or depth of the first flow path 110 and the second flow path 120 is, for example, about several μm to several hundred mm, several μm to several tens mm, several μm to several mm, It is also good.

By using such a separation unit, for example, plasma components can be rapidly separated from blood. In addition, as described later, it is easy to make the blood cells be less likely to be clogged in the filter, and hemolysis can be less likely to occur. Also, there is no need to learn special skills to perform operations. In addition, since the separation unit can be disposable, the risk of contamination between specimens, infection due to pathogenic microorganisms and the like contained in the specimens is reduced. In addition, it can cope with a small amount of sample.

(Modification)
In the separation unit 100 described above, the pump unit 130 is disposed only in the first flow passage 110. However, the pump unit may be disposed only in the second flow passage 120 or may be disposed in both the first flow passage 110 and the second flow passage 120.

FIG. 2 is a schematic view illustrating a fluid device in which the fluid control unit is disposed in both of the first fluid introducing unit 110 and the second fluid introducing unit 120. As shown in FIG. 2, the fluid device of the present modification has a configuration similar to that of the above-described separation unit 100 except that it further includes a fluid adjustment unit 135 in addition to the fluid adjustment unit 130. The fluid regulator 135 regulates the flow of fluid in the second fluid inlet 120.

FIG. 2 can also be said to be a schematic view showing an example of the separation part of the fluid device of this modification. As shown in FIG. 2, the separation unit 200 has the same configuration as the separation unit 100 except that the separation unit 200 further includes a pump unit (fluid adjustment unit) 135 in addition to the pump unit (fluid adjustment unit) 130. The pump unit 135 regulates the flow of fluid in the second flow passage (second fluid introduction unit) 120.

In the separation of the substance to be separated using the separation unit 200, the pump unit 135 can also be operated in addition to the pump unit 130. As a result, not only the first fluid in the first channel 110 but also the second fluid in the second channel 120 can be circulated, and the fluid in the fluid inside the first channel 110 can be circulated. The time until the concentration of the substance to be separated and the concentration of the substance to be separated in the fluid inside the second flow channel 120 reach equilibrium can be further shortened.

Here, the direction of the flow of the first fluid in the first flow passage 110 circulated by the pump unit 130 and the direction of the flow of the second fluid in the second flow passage 120 circulated by the pump unit 135 are It is not particularly limited. The flow of fluid may be in the same direction or in different directions on the first flow path side of the filter 140 and on the second flow path side of the filter 140.

In the example described above, the fluid introduced into the inside of the first channel 110 and the second channel 120 is a liquid. However, the fluid introduced into the inside of the first flow passage 110 and the second flow passage 120 is not limited to liquid, and may be gas. In this case, examples of the substance to be separated include fine particles such as dust, bacteria and viruses. When such a substance is separated, for example, a fluorine resin filter or the like can be used as the filter 140.

Hereinafter, each configuration of the separation unit of the present embodiment will be described.
(Pump part (fluid control part))
The

pump unit

130 or 135 may have three or more valves. Also, the above-mentioned valve is a diaphragm valve provided with a diaphragm member, and the diaphragm member may be formed of an elastomeric material.

FIGS. 3A and 3B are cross-sectional views for explaining an example of the structure of the diaphragm valve. 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, the diaphragm valve 300 includes a first substrate 310, a diaphragm member 330 made of an elastomeric material, and a second substrate 320. The second substrate 320 and the diaphragm member 330 are adhered in close contact with each other. Also, the space between the first substrate 310 and the diaphragm member 330 forms a flow path 315 through which the fluid flows. In addition, a through hole 340 is provided in part of the second substrate 320. Further, in the through hole 340, the diaphragm member 330 is exposed.

The diaphragm valve 300 is disposed in the flow passage 315 and regulates the flow of fluid inside the flow passage 315.

In the open state of the diaphragm valve 300 shown in FIG. 3A, the fluid can flow in the flow path 315. On the other hand, as shown in FIG. 3B, when a fluid for valve control is supplied from the through hole 340 of the diaphragm valve 300 and the inside of the through hole 340 is pressurized, the diaphragm member 330 is deformed and the diaphragm member is deformed. A portion of the substrate 330 is in close contact with the first substrate 310. This state is a closed state of the diaphragm valve 300. As a result, the flow of fluid inside the flow path 315 is shut off.

Here, as shown in FIGS. 3A and 3B, in order to make the closed state of the diaphragm valve 300 stronger, a convex portion is formed in the region of the first substrate 310 facing the through hole 340. 311 may be formed.

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 340.

The elastomer material forming the diaphragm member 330 is not particularly limited as long as it is a material that can be deformed in the axial direction of the through hole 340 according to the pressure change inside the through hole 340, for example, polydimethylsiloxane (PDMS) And silicone-based elastomers such as polymethylphenylsiloxane and polydiphenylsiloxane.

In the diaphragm valve 300 in the closed state shown in FIG. 3B, when the pressure applied to the inside of the through hole 340 is reduced, the deformed diaphragm member 330 returns to its original shape, and again FIG. It will be in the open state shown in). As a result, the fluid can flow again inside the flow path 315.

A pump can be formed by three or more valves, that is, by arranging three or more valves. FIGS. 4A to 4D are cross-sectional views for explaining the operation of an example of a pump provided with three

diaphragm valves

300a, 300b and 300c.

In the state shown in FIG. 4A, all the three

valves

300a, 300b and 300c are in the open state. In this state, since the flow of fluid inside the flow path 315 is not controlled, the fluid may flow to the right or to the left as shown in FIG. It may be stopped.

Subsequently, as shown in FIG. 4 (b), the valve 300a is controlled to be closed, and the

valves

300b and 300c are kept open. As a result, the flow of fluid inside the flow path 315 is blocked by the valve 300a.

Subsequently, as shown in FIG. 4C, the valve 300a and the valve 300b are controlled to be closed, and the valve 300c is kept open. Here, in the process of the valve 300b changing from the open state to the closed state, the diaphragm member 330 is deformed, so that the fluid existing around the valve 300b is pushed away. However, since the valve 300a is in the closed state, the displaced fluid moves in the direction indicated by the arrow in FIG. 4C, that is, to the right in FIG. 4C. As a result, the fluid has a flow in the direction indicated by the arrow.

Subsequently, as shown in FIG. 4 (d), the

valves

300b and 300c are controlled to be closed. Then, in the process of changing the valve 300 c from the open state to the closed state, the diaphragm member 330 is deformed, so that the fluid existing around the valve 300 c is pushed away. However, since the valve 300a is in the closed state, the displaced fluid moves in the direction shown by the arrow in FIG. 4 (d), that is, to the right in FIG. 4 (d). As a result, the flow in the direction indicated by the arrows in the fluid is further accelerated. At this time, the valve 300a may be controlled to be open as shown in FIG. 4 (d), or may be kept closed.

Subsequently, as shown in FIG. 4A again, all the

valves

300a, 300b and 300c are controlled to be in the open state. Here, due to inertia, the fluid in the flow path 315 may continue to move to the right toward FIG. 4A.

Furthermore, by repeating the above steps, the flow of fluid inside the flow path 315 can be controlled.

The above process is an example of a control method of a pump provided with three valves, and a control method of a pump provided with three valves is not limited to this. For example, in the control of the valves described above, the flow of fluid inside the flow path 315 can be controlled in the reverse direction to that described above by reversing the timing of opening and closing the

valves

300a and 300c. Further, by adjusting the cycle of repeating the operation of FIG. 4 (a) to FIG. 4 (d), a pulsating micro solution flow is formed to suppress clogging of the filter 140 with foreign substances, or separation speed is reduced. It can also be controlled. The flow of the solution through the filter 140 can also be controlled by adjusting the pressure of the gas driving the valves 300a to 300c and the diameter of the valves. Thereby, clogging of the filter 140 may be suppressed, or the separation speed may be controlled.

Moreover, it is also possible to control the flow of the fluid inside the flow path 315 by a pump provided with four or more valves.

Also, the structure of the diaphragm valve is not limited to that described above. For example, as a diaphragm valve, a tubular structure having an outer cylindrical portion and an inner cylindrical portion described in International Publication No. 2016/006615, and a thin film portion arranged to cover one end of the inner cylindrical portion It is also possible to use a valve provided with a diaphragm member that has an anchor portion that goes around the periphery of the thin film portion and adheres along the inner wall of the outer cylinder portion and the outer wall of the inner cylinder portion.

FIGS. 5 (a) and 5 (b) are cross-sectional views for explaining the structure of the diaphragm valve described in WO 2016/006615. 5A shows the valve 500 in the open state, and FIG. 5B shows the valve 500 in the closed state.

As shown in FIGS. 5A and 5B, in the valve 500, compared with the valve 300 described above, the diaphragm member 330 is not disposed on the entire surface of the second substrate 320, and only around the valve 500. The points that are locally arranged are mainly different.

Since the diaphragm member 330 of the valve 500 includes the anchor portion 331, the diaphragm member 330 is not broken and does not peel off from the second substrate 320 even when the diaphragm member 330 is deformed by pressure. .

In the separation part of the present embodiment, as another usable valve, for example, raising a two-color molded valve obtained by continuously molding a resin part and an elastomer part using two molds Can. The two-color molded valve has an advantage that the time required for production is short and the adhesion between the resin and the elastomer is high.

Although the example in which the pump unit is disposed in the first channel 110 and the second channel 120 has been described above, the pump unit is not necessarily disposed in the first channel or the second channel. You do not have to. For example, when the second flow passage 120 is not in a closed circuit, a reservoir for supplying the second fluid is connected to one end of the second flow passage, and the other fluid contains the substance to be separated at the other end. The second fluid can be moved into the second flow passage 120 by connecting a reservoir for recovering the fluid, and providing a suction unit in the reservoir for suction.

(filter)
In the separation unit of the present embodiment, as an example, the filtering direction of the filter 140 in the filter 140 is substantially orthogonal to the flow direction of the fluid in the first flow passage 110 and the flow direction of the fluid in the second flow passage 120 It is arranged to be. In the present specification, the filtration direction of the filter 140 means the direction in which the substance to be separated passes through the filter 140.

As shown in FIGS. 1 and 2, in the separation unit 100 and the separation unit 200, the filtration direction of the filter 140 is the direction in which the fluid flows in the first flow passage 110 and the fluid flows in the second flow passage 120. It is orthogonal to the direction. Thereby, it is possible to prevent the substances other than the substance to be separated contained in the first fluid from being clogged by the first fluid directly in the filtration direction of the filter 140. The substances other than the substance to be separated mean, for example, red blood cells, white blood cells, platelets and the like in the case of separating plasma components from blood.

In the separation unit of the present embodiment, the filter may be a screen filter or a depth filter. The screen filter is a filter whose pore diameter is accurately determined, and has a structure in which cylindrical pores are uniformly distributed, and an object is captured on the filter surface. A depth filter is a filter in which objects are captured on the surface of the filter and inside the filter.

The filter of the present embodiment may be a membrane filter. The membrane filter is a porous membrane with uniform pore diameter, and those made of fluorocarbon resin, cellulose acetate, polycarbonate resin, etc. are commercially available. The membrane filter is a type of screen filter that captures an object on the surface, and filters of different pore sizes are selected according to the purpose. By using a membrane filter as the filter, separation of the substance to be separated based on the pore size can be performed more accurately.

In the separation portion of the present embodiment, the wall surfaces of the first flow path 110 and the second flow path 120 and the peripheral edge of the filter 140 are elastomers composed of the same components as the elastomer material constituting the diaphragm member 330 of the diaphragm valve. It may be fixed by a material.

The separation unit including the diaphragm valve 300 includes, for example, a first substrate 310 made of resin in which a flow passage and a through hole 340 for forming a valve are formed, a diaphragm member 330 made of a sheet of elastomer material, and a flow passage and valve It can manufacture by laminating | stacking and thermocompression-bonding the 2nd board | substrate 320 which consists of resin in which the through-hole 340 for formation was formed.

Further, for example, a groove (concave portion) to be a part of the first flow passage 110 is formed on the surface of the first substrate 310, and a groove (concave portion) to be a part of the second flow passage 120 is When formed on the surface of the substrate 320 and the flow paths formed in these grooves communicate with each other through the filter 140, the diaphragm member 330 turns a portion corresponding to the filter 140 a little smaller than the filter 140. Keep it through. Then, the first substrate 310, the diaphragm member 330, the filter 140, and the second substrate 320 are stacked and thermocompression-bonded, etc., so that the filter 140 includes the diaphragm member 330 and the second substrate 320. It can be fixed in between.

FIG. 6A is a perspective view for explaining an example of the separation unit 200. FIG. 6 (b) is a cross-sectional view taken along the line b-b 'in FIG. 6 (a). As shown in FIGS. 6A and 6B, on the surface of the first substrate 310, a groove to be a part of the first flow path 110 is formed. Further, on the surface of the second substrate 320, a groove to be a part of the second flow passage 120 is formed.

In the separation unit 200, the first flow passage 110 and the second flow passage 120 formed by the grooves are configured to communicate with each other through the filter 140.

The diaphragm member 330 is cut out slightly smaller than the filter 140 at a portion where the diaphragm member 330 and the filter 140 are in contact with each other. As a result, the peripheral portion of the cut out portion of the diaphragm member 330 and the peripheral portion of the filter 140 overlap and are in contact with each other. The first substrate 310, the diaphragm member 330, the filter 140, and the second substrate 320 are stacked in this order and bonded by thermocompression bonding or the like.

Thus, the filter 140 is fixed between the diaphragm member 330 and the second substrate 320. As described later in the embodiment, the inventors firmly fix the filter 140 with such a structure without leakage of the fluid flowing inside the first flow passage 110 and the second flow passage 120. I clarified that I could do it.

The order in which the filter 140 and the diaphragm member 330 are stacked is not limited to this. For example, the first substrate 310, the filter 140, the diaphragm member 330, and the second substrate 320 are sequentially stacked and bonded by thermocompression bonding or the like. It may be done.

[Method of separating separated substances]
The method for separating a substance to be separated according to the present embodiment is a method for separating a substance to be separated from a first fluid containing the substance to be separated using a fluid device, wherein the fluid device contains the first fluid A first fluid inlet to be introduced, a second fluid inlet to which a second fluid for recovering the substance to be separated is introduced, a flow of the first fluid, and / or a flow of the second fluid And at least one of the first fluid introduction portion and the second fluid introduction portion has a flow path shape, and at least one of the first fluid introduction portion and the second fluid introduction portion. A part is a filter that selectively transmits the substance to be separated, or a fluid device in communication with each other via a filter that does not selectively transmit substances other than the substance to be separated, and the second part of the separation unit (1) The first fluid is introduced into the fluid introducing portion Introducing, and introducing the second fluid into the second fluid inlet, and activating the fluid regulator that regulates the flow of the first fluid and / or the flow of the second fluid. Moving the first fluid and / or the second fluid, and recovering the fluid in the second fluid inlet.

In the separation method of the substance to be separated according to the present embodiment, a first fluid containing the substance to be separated is introduced into the first flow path of the separation unit of the fluid device described above, and the object to be separated is introduced into the second flow path. A step of introducing a second fluid that recovers separated substances, a pump unit that regulates the flow of the first fluid, and / or a pump unit (fluid regulation unit) that regulates the flow of the second fluid It may be said that the method comprises the steps of: moving the first fluid and / or the second fluid; and recovering the substance to be separated together with the second fluid.

In the fluid device, the first fluid introducing portion and the second fluid introducing portion may both have a flow path shape and form a closed circuit. Also, the fluid control unit for controlling the flow of the first fluid and the second fluid may be disposed in the first fluid introducing unit and the second fluid introducing unit, respectively, and may include three or more series of valves. Good.

Also, the first fluid and the second fluid may move in opposite directions around the first fluid inlet and the second fluid inlet, respectively.

By the separation method of the present embodiment, for example, plasma components can be rapidly separated from blood. In addition, the filter of the separation unit is disposed such that the filtration direction of the filter is substantially orthogonal to the flow direction of the fluid in the first flow path and the flow direction of the fluid in the second flow path. It is possible to prevent the separation substance from being directly subjected to force in the filtration direction and clogging the filter. Thus, for example, blood cells can be prevented from clogging the filter, and hemolysis is unlikely to occur.

Moreover, the separation method of the present embodiment does not need to learn a special technique to perform the operation. In addition, since the separation unit can be disposable, the risk of contamination between specimens, infection due to pathogenic microorganisms and the like contained in the specimens is reduced. In addition, it can cope with a small amount of sample.

Next, the present embodiment will be described by way of examples, but the present invention is not limited to the following examples.

[Experimental Example 1]
(Manufacturing of separation part)
A fluidic device having a separator shown in FIG. 7 is produced. The separation unit includes a first flow passage 110, a second flow passage 120, a pump unit 130 disposed in the first flow passage, and a pump unit 135 disposed in the second flow passage. It was Further, parts of the first flow path 120 and the second flow path 120 were in communication with each other via the filter 140.

Further, the inflow channel 111 and the discharge channel 112 were connected to the first channel 110. In addition, the separation unit controls the communication between the inflow channel 111 and the first channel 110 in the open or closed state, and the communication between the discharge channel 112 and the first channel 110. And a valve 114 for controlling the open and close states.

Further, the inflow channel 121 and the discharge channel 122 were connected to the second flow channel 120. In addition, the separation unit controls the communication between the inflow channel 121 and the second channel 120 in the open or closed state, and the communication between the discharge channel 122 and the second channel 120. And the valve 124 for controlling the open state or the closed state.

[Experimental Example 2]
(Separation using separation unit)
The fluid containing the substance to be separated was introduced into the separation part manufactured in Experimental Example 1 to separate the substance to be separated.

As a fluid containing a substance to be separated, colored water in which beads were dispersed was used. This is an imitation of blood, and beads are assumed to be blood cells such as red blood cells and white blood cells, and may be hereinafter referred to as "simulated blood". As colored water, a phosphate buffer (PBS) in which a dye was dissolved was used. In the present experimental example, the substance to be separated corresponds to the pigment. In addition, PBS containing no dye was used as a fluid for recovering the substance to be separated.

First, simulated blood was introduced into the first channel 110, and PBS was introduced into the second channel. FIG. 8 is a photograph of the separation part at this stage. The upper left "00:05" indicates that 5 seconds after the start of the experiment.

Subsequently, the

pump portions

130 and 135 were operated to flow the fluid in the first flow passage 110 and the fluid in the second flow passage 120. Thereby, the pigment in the simulated blood in the first channel permeates through the filter 140, moves into the PBS in the second channel 120, and is separated from the simulated blood. On the other hand, since the beads can not permeate through the filter 140, they remain in the fluid in the first channel 110. In addition, by operating the pump unit, the time for the dye movement to reach equilibrium was shortened.

FIG. 9 is a photograph of the separation portion about 250 seconds after the start of the experiment. The upper left "04: 10" indicates 4 minutes and 10 seconds after the start of the experiment (after 250 seconds). In FIG. 9, it can be seen that the concentration of the dye reached equilibrium, and the concentration of the dye in the first channel and the concentration of the dye in the second channel became almost the same.

FIG. 10 is a graph showing the results of measuring the concentration of the dye in the second channel over time. As a result, it was revealed that it took 245 seconds for the dye to reach an equilibrium arrival rate of 90% when the dye was separated from the mock blood.

The equilibrium arrival rate of 90% was determined as follows. That is, as a result of measuring the concentration of the dye in the second channel by light detection, the fluctuation of the light intensity is almost eliminated, and the light intensity at equilibrium is balanced with the light intensity after 701 seconds to 900 seconds from the start of the experiment The point at which the light intensity of 90% of the above was reached was taken as the equilibrium arrival rate of 90%.

100, 200 ... separation unit, 110 ... first flow passage (first fluid introduction unit), 120 ... second flow passage (second fluid introduction unit), 111, 121 ... inflow passage, 112, 122 ...

Discharge channels

113, 114, 123, 124: valves, 130, 135: pump parts (fluid control parts), 140: filters, 300, 300a, 300b, 300c, 500: diaphragm valves, 310:

first substrate

311, 311a, 311b, 311c: convex portion, 315: flow path, 320: second substrate, 330: diaphragm member, 331: anchor portion, 340, 340a, 340b, 340c ... through hole.

Claims

A first fluid introducing portion into which a first fluid containing a substance to be separated is introduced;
A second fluid introducing unit into which a second fluid for recovering the substance to be separated is introduced;
A fluid control unit that controls the flow of the first fluid and / or the flow of the second fluid;
At least one of the first fluid introduction part and the second fluid introduction part has a flow path shape,
At least a part of the first fluid introduction part and the second fluid introduction part is a filter that selectively transmits the substance to be separated, or a filter that does not selectively transmit a predetermined substance other than the substance to be separated. Fluid devices in communication with each other.
The fluid device according to claim 1, wherein the first fluid introducing portion and / or the second fluid introducing portion having the flow path shape form a closed circuit.
The fluid control units for controlling the flow of the first fluid or the second fluid are respectively disposed in the first fluid introducing unit and / or the second fluid introducing unit having the flow path shape, and The fluidic device according to claim 2, comprising three or more valves.
The fluidic device according to claim 3, wherein the valve is a diaphragm valve comprising a diaphragm member, the diaphragm member consisting of an elastomeric material.
Including at least two substrates bonded together,
A first recess that is at least a part of a first fluid introduction portion is formed on the surface of the second substrate facing the first substrate,
A second recess that is at least a part of a second fluid introduction portion is formed on the surface of the first substrate facing the second substrate,
The filter is sandwiched between the first substrate and the second substrate,
The fluid device according to any one of claims 1 to 4, wherein the first recess and the second recess face each other through the filter.
The fluid device according to claim 5, wherein the diaphragm member is an elastomer sheet, and the elastomer sheet is sandwiched between the first substrate and the second substrate.
The fluid device according to any one of the preceding claims, wherein the first fluid is a blood sample.
Each of the first fluid introduction part and the second fluid introduction part has a flow channel shape,
The filter according to any one of claims 1 to 7, wherein a filtration direction of the filter is substantially orthogonal to a direction in which the fluid flows through the first fluid inlet and a direction in which the fluid flows through the second fluid inlet. The fluidic device according to claim 1.
Each of the first fluid introduction part and the second fluid introduction part has a flow channel shape,
The fluid according to any one of claims 1 to 8, further comprising a portion in which the first fluid introducing portion and the second fluid introducing portion are substantially parallel, and the filter is provided in the parallel portion. device.
The fluid device according to any one of the preceding claims, wherein the filter is a membrane filter.
A method of separating a substance to be separated from a first fluid containing the substance to be separated using a fluid device, comprising:
The fluidic device is
A first fluid introducing portion into which the first fluid is introduced;
A second fluid introducing unit into which a second fluid for recovering the substance to be separated is introduced;
A fluid control unit that controls the flow of the first fluid and / or the flow of the second fluid;
At least one of the first fluid introduction part and the second fluid introduction part has a flow path shape,
At least a part of the first fluid introducing portion and the second fluid introducing portion communicate with each other through a filter that selectively transmits the substance to be separated or a filter that does not selectively transmit other than the substance to be separated The fluid device being
Introducing the first fluid into the first fluid inlet of the separation unit, and introducing the second fluid into the second fluid inlet;
Activating a fluid regulator that regulates the flow of the first fluid and / or the flow of the second fluid to move the first fluid and / or the second fluid;
Recovering the fluid in the second fluid inlet;
A method of separating a substance to be separated, including
Each of the first fluid introducing portion and the second fluid introducing portion has a flow path shape and forms a closed circuit, and regulates the flow of the first fluid and the second fluid. The method for separating a substance to be separated according to claim 11, wherein the fluid control unit is disposed in each of the first fluid introducing unit and the second fluid introducing unit, and includes three or more valves each.
The method according to claim 12, wherein the first fluid and the second fluid respectively move around the first fluid inlet and the second fluid inlet in opposite directions.