Sample solution separation device, sample solution separation system, and sample solution separation method

Abstract

The invention provides a sample solution separation device with a small dead volume. A sample solution separation device (101) provided with: a plurality of microcells (105); a flow path (104) for connecting the plurality of microcells (105); a first opening (102) as an inlet for introducing a sample solution into the flow path (104); a valve (103) provided between the first opening (102) and the plurality of microcells (105); and a solid phase (106) provided between the second opening (107) and the plurality of microchambers (105), the solid phase (106) having air permeability, having water repellency to the sample solution, and having the ability to pass through the separation liquid, the second opening (107) being provided on the opposite side of the first opening with the plurality of microchambers therebetween, and the solid phase (106) being provided between the second opening (107) and the plurality of microchambers (105).

Description

Technical Field

[0001] The invention relates to a sample solution separation device, a sample solution separation system and a sample solution separation method for separating a sample solution. Background Art

[0002] Digital PCR (Polymerase Chain Reaction) is a technology for detecting nucleic acids with high sensitivity. Compared with conventional real-time quantitative PCR, it can detect low-frequency gene mutations.

[0003] Digital PCR involves dividing a sample solution into tiny volumes, then performing PCR within the divided solutions to amplify and measure the DNA being measured. Patent Document 1 discloses a method for placing a sample solution in a tiny container, sealing the port to evacuate the container, and then introducing the sample solution into the container. Non-Patent Document 1 also discloses a method for degassing the container, closing the outlet valve, and opening the inlet valve to add the solution. Patent Document 2 discloses a structure that uses a gas-liquid separation filter to dispense the solution without waste.

[0004] Prior art literature

[0005] Patent Literature

[0006] Patent Document 1: U.S. Patent No. 11305276

[0007] Patent Document 2: Japanese Patent No. 4888773

[0008] Non-patent literature

[0009] Non-patent literature 1: Micromachines 2020, 11, 1025; doi:10.3390 / mi11121025 Summary of the Invention

[0010] Problems to be solved by the invention

[0011] In digital PCR, a sample solution containing the target DNA is separated into multiple regions, and PCR is performed on each region to identify the type of DNA present in each region. The characteristic is that by performing PCR after separation into each region, the target DNA can be measured with high sensitivity. For example, liquid biopsy for detecting trace amounts of cell-free DNA (cfDNA) in the blood is one of the suitable applications. Cell-free DNA sometimes contains ctDNA (circulating tumor DNA) which is tumor-derived DNA. It is expected that by measuring the types and ratios of mutations in ctDNA, it can be applied to cancer diagnosis, treatment selection, and monitoring of treatment effects.

[0012] cfDNA and ctDNA are present in trace amounts in blood, necessitating high-sensitivity measurements. In digital PCR, the sample solution is separated into multiple, micro-volume chambers for measurement. This process prevents the entire sample solution from being used, resulting in a dead volume of solution that cannot be separated into the micro-volume chambers and, therefore, cannot be measured. To achieve high-sensitivity measurements, it is necessary to measure as much of the collected sample solution as possible. In other words, the dead volume of the sample solution needs to be reduced.

[0013] The present invention has been made in view of the above circumstances, and one object thereof is to provide a sample solution separation device, a sample solution separation system, and a sample solution separation method capable of reducing the dead volume of a sample solution.

[0014] Methods for solving problems

[0015] In order to solve the above-mentioned problems, the sample solution separation device of the present invention is a sample solution separation device for separating a sample solution, which comprises: a plurality of microchambers; a flow path connecting the plurality of microchambers; a first opening serving as an inlet for the sample solution to enter the flow path; a valve arranged between the first opening and the plurality of microchambers; a second opening serving as an inlet for a separation liquid to enter the flow path, and arranged on the opposite side of the first opening across the plurality of microchambers, the separation liquid separating the plurality of microchambers containing the sample solution; and a solid phase arranged between the second opening and the plurality of microchambers, the solid phase having air permeability, water repellency (hydrophobicity) to the sample solution, and permeability to the separation liquid.

[0016] In addition, the sample solution separation system of the present invention has a sample solution separation device, a pump, a valve control unit and a separation liquid introduction unit. The sample solution separation device separates the sample solution and is equipped with: a plurality of microchambers; a flow path connecting the plurality of microchambers; a first opening serving as an inlet for the sample solution to enter the flow path; a valve disposed between the first opening and the plurality of microchambers; a second opening serving as an inlet for the separation liquid to enter the flow path and disposed on the opposite side of the first opening across the plurality of microchambers, the separation liquid separating the plurality of microchambers containing the sample solution; and a solid phase disposed between the second opening and the plurality of microchambers, the solid phase being breathable, water-repellent to the sample solution, and permeable to the separation liquid. The pump degases the air in the plurality of microchambers and the flow path from the second opening through the solid phase. The valve control unit opens the valve. The separation liquid introduction unit introduces the separation liquid from the second opening.

[0017] In addition, the sample solution separation method of the present invention has the following characteristics:

[0018] A process of preparing a sample solution separation device for separating a sample solution, the sample solution separation device comprising: a plurality of microchambers; a flow path connecting the plurality of microchambers; a first opening serving as an inlet for the sample solution to enter the flow path; a valve disposed between the first opening and the plurality of microchambers; a second opening serving as an inlet for a separation liquid to enter the flow path, the valve being disposed on the opposite side of the first opening across the plurality of microchambers, the separation liquid separating the plurality of microchambers containing the sample solution; and a solid phase disposed between the second opening and the plurality of microchambers, the solid phase being air permeable, water repellent to the sample solution, and permeable to the separation liquid.

[0019] The process of degassing the air in multiple micro-chambers and flow channels through the solid phase,

[0020] The valve is opened to introduce the sample solution from the first opening into the plurality of microchambers and flow paths, and

[0021] The process of introducing the separation liquid into the flow channel from the second opening through the solid phase.

[0022] Effects of the Invention

[0023] According to the sample solution separation device, the sample solution separation system, and the sample solution separation method of the present invention, the dead volume of the sample solution can be reduced when the sample solution is separated into a plurality of microchambers.

[0024] Other objects and novel features will become apparent from the description of this specification and the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 1 is a structural diagram of the sample solution separation device 101 of Example 1.

[0026] Figure 2A 1 is a diagram showing a state where a sample solution 201 and a separation liquid 202 are introduced into the sample solution separation device 101 of Example 1. FIG.

[0027] Figure 2B 1 is a diagram showing a state where a sample solution 201 and a separation liquid 202 are introduced into the sample solution separation device 101 of Example 1. FIG.

[0028] Figure 2C 1 is a diagram showing a state where a sample solution 201 and a separation liquid 202 are introduced into the sample solution separation device 101 of Example 1. FIG.

[0029] Figure 2D 1 is a diagram showing a state where a sample solution 201 and a separation liquid 202 are introduced into the sample solution separation device 101 of Example 1. FIG.

[0030] Figure 3This is a flow chart for introducing a sample solution 201 and a separation solution 202 into the sample solution separation device 101 of Example 1 and separating the sample solution 201 into the microchamber 105 .

[0031] Figure 4 1 is a structural diagram of a sample solution separation system 400 that separates a sample solution 201 and introduces the sample solution 201 into the sample solution separation device 101 of Example 1.

[0032] Figure 5 4 is a structural diagram of a sample solution separation device 501 according to Example 2.

[0033] Figure 6A 3 is a top view of the sample solution separation device 601 of Example 3.

[0034] Figure 6B It is a side view of the sample solution separation device 601 of Example 3.

[0035] Figure 7 1 is a structural diagram of a sample solution separation system 700 for separating a sample solution 201 and introducing the sample solution 201 into a sample solution separation device 601 of Example 3.

[0036] Figure 8 4 is a structural diagram of the sample solution separation device 801 of Example 4.

[0037] Figure 9 1 is a structural diagram of the digital PCR system 900 of Example 5.

[0038] Figure 10 This is an operation flow chart of the digital PCR system 900 of Example 5. DETAILED DESCRIPTION

[0039] In the following embodiments, for convenience, when necessary, they are divided into multiple parts or embodiments for description. However, unless otherwise specified, they are not mutually exclusive, but rather one part is a modification, detail, supplementary explanation, etc. of part or all of the other. In addition, in the following embodiments, when the number of elements, etc. (including number, value, amount, range, etc.) is mentioned, except where otherwise specified or where it is clearly limited to a specific number in principle, it is not limited to the specific number and can be above or below the specific number.

[0040] Furthermore, in the following embodiments, references to constituent elements (including element steps, etc.) are not essential, unless otherwise specifically stated or clearly considered essential in principle. Similarly, in the following embodiments, references to the shapes, positional relationships, etc. of constituent elements include references to substantially similar or similar shapes, etc., unless otherwise specifically stated or clearly considered essential in principle. The same applies to the numerical values ​​and ranges described above.

[0041] In addition, in all the drawings for describing the embodiments, the same reference numerals are attached to the same components in principle, and their repeated descriptions are omitted.

[0042] (Example 1)

[0043] use Figures 1 to 4 The sample solution separation device 101 of Example 1 will be described. Figure 1 This is a structural diagram of a sample solution separation device 101 according to Example 1. The sample solution separation device 101 is a device that separates and stores a sample solution into a plurality of microchambers 105. The sample solution separation device 101 includes a first opening 102, a valve 103, a flow path 104, a plurality of microchambers 105, a solid phase 106, and a second opening 107.

[0044] The flow path 104 is connected to the first opening 102 via the valve 103. The flow path 104 is connected to the plurality of microchambers 105. The flow path 104 is also connected to the second opening 107 via the solid phase 106. That is, the flow path 104 connects the first opening 102, the plurality of microchambers 105, and the second opening 107.

[0045] The first opening 102 is an inlet for introducing the sample solution into the flow channel 104 .

[0046] The valve 103 is provided between the first opening 102 and the plurality of microchambers 105 .

[0047] The second opening 107 is an inlet for introducing a separation solution that separates the plurality of microchambers 105 containing the sample solution into the flow channel 104. The second opening 107 is provided on the opposite side of the first opening 102 with the plurality of microchambers 105 interposed therebetween.

[0048] The solid phase 106 is provided between the second opening 107 and the plurality of microchambers 105. The solid phase 106 has air permeability, is water-repellent to the sample solution, and is permeable to the separation solution.

[0049] Figures 2A to 2D 1 is a diagram showing a state in which a sample solution 201 and a separation liquid 202 are introduced into the sample solution separation device 101 of Example 1. FIG.

[0050] First, the sample solution 201 is introduced into the first opening 102. When the sample solution 201 is introduced into the first opening 102, the valve 103 is closed ( Figure 2A After the valve 103 is closed, the air inside the flow path 104 and the microchamber 105 is degassed from the second opening 107 through the solid phase 106. That is, the flow path 104 and the microchamber 105 are brought into a vacuum state.

[0051] By making the flow path 104 and the microchamber 105 into a vacuum state and opening the valve 103, the sample solution 201 is introduced into the flow path 104 and the microchamber 105 ( Figure 2B ).

[0052] Here, solid phase 106 is a solid having air permeability, hydrophobicity, and lipophilicity. The air permeability of solid phase 106 allows air to pass through, and as described above, enables the flow path 104 and microchamber 105 to be in a vacuum state. Furthermore, the hydrophobicity of solid phase 106 repels aqueous solutions such as sample solution 201, thereby preventing the passage of sample solution 201.

[0053] Therefore, the sample solution 201 introduced into the flow path 104 stops at the moment of reaching the solid phase 106. Since the microchamber 105 is in a vacuum state, the sample solution 201 continues to be introduced. Thus, the microchamber 105 is filled with the sample solution 201 ( Figure 2C ).

[0054] The separation solution 202 (see Figure 2D ) A substance (eg, oil) having a physical property of not being mixed with the sample solution 201 is used. The solid phase 106 has lipophilicity, thereby allowing the separation liquid 202 to pass therethrough.

[0055] Next, the sample solution 201 in the flow channel 104 is replaced by the separation liquid 202. The separation liquid 202 is introduced from the second opening 107. The solid phase 106 is lipophilic and has the property of allowing the separation liquid 202 to pass through. Therefore, the separation liquid 202 is introduced from the second opening 107 through the solid phase 106 into the flow channel 104. The sample solution 201 in the flow channel 104 is pushed back toward the first opening 102, and the flow channel 104 is filled with the separation liquid 202 ( Figure 2D At this time, the sample solution 201 remains in the microchamber 105 , and thus the sample solution 201 in the plurality of microchambers 105 is separated by the separation liquid 202 in the flow channel 104 .

[0056] In this manner, by using the air-permeable, hydrophobic, and lipophilic solid phase 106, air can be expelled from the flow path 104 and the microchamber 105, and the sample solution 201 can be stopped at the position of the solid phase 106. In addition, the solid phase 106 can introduce the separation liquid 202 into the flow path 104, divide the sample solution 201, and seal it in the microchamber 105.

[0057] Figure 3 This is a flow chart for introducing a sample solution 201 and a separation solution 202 into the sample solution separation device 101 of Example 1 and separating the sample solution 201 into the microchamber 105 .

[0058] First, the sample solution 201 is placed in the first opening 102 ( S301 ), and the valve 103 closes the flow path 104 .

[0059] Then, by driving the pump 403 connected to the second opening 107 (see Figure 4 ), the air in the flow path 104 and the plurality of microchambers 105 is exhausted through the solid phase 106 and the second opening 107 (S302). As a result, the flow path 104 and the plurality of microchambers 105 are in a vacuum state.

[0060] After the flow channel 104 and the plurality of microchambers 105 are in a vacuum state, the valve 103 opens the flow channel 104 , whereby the sample solution 201 is introduced from the first opening 102 through the valve 103 into the flow channel 104 and the plurality of microchambers 105 ( S303 ).

[0061] Then, by driving the pump 405 connected to the second opening 107 (see Figure 4 ), the separation liquid 202 is introduced into the flow channel 104 via the second opening 107 and the solid phase 106 (S304). The sample solution 201 in the flow channel 104 is pushed back toward the first opening 102 by the separation liquid 202, and the sample solution 201 in the flow channel 104 is replaced by the separation liquid 202, and the plurality of microchambers 105 are separated by the separation liquid 202. It should be noted that after the separation liquid 202 is introduced into the flow channel 104, the valve 103 may be closed to prevent the sample solution 201 in the plurality of microchambers 105 and the separation liquid 202 in the flow channel 104 from moving.

[0062] Figure 4 This is a structural diagram of a sample solution separation system 400 that separates a sample solution 201 and introduces the sample solution into the sample solution separation device 101 of Example 1. The sample solution separation system 400 includes a valve control mechanism 401, an opening contact portion 402, a pump 403, an opening contact portion 404, a pump 405, a moving mechanism 406, and the sample solution separation device 101.

[0063] The valve control mechanism 401 (valve control unit) controls the opening and closing of the valve 103. When the sample solution 201 is introduced into the plurality of microchambers 105, the valve control mechanism 401 opens the valve 103 from the closed state.

[0064] The opening contact portion 402 is connected to the second opening 107 of the sample solution separation device 101. Furthermore, the opening contact portion 402 is connected to a pump 403 for evacuating the flow path 104 and the plurality of microchambers 105. The moving mechanism 406 moves the opening contact portion 402 to connect to the second opening 107 when evacuating the flow path 104 and the plurality of microchambers 105.

[0065] The pump 403 degases the air in the flow path 104 and the microchambers 105 from the second opening 107 through the solid phase 106 , thereby vacuuming the flow path 104 and the plurality of microchambers 105 .

[0066] The opening contact portion 404 is connected to the second opening 107 of the sample solution separation device 101. In addition, the opening contact portion 404 is connected to a pump 405 that introduces the separation liquid 202 into the flow channel 104.

[0067] The moving mechanism 406 (switching unit) connects the opening contact portion 402 to the second opening 107 when degassing air from the flow channel 104 and the plurality of microchambers 105. Furthermore, when introducing the separation liquid 202 into the flow channel 104, the moving mechanism 406 moves the opening contact portion 404 to connect it to the second opening 107. In other words, the moving mechanism 406 switches the connection destination of the second opening 107 to either the pump 403 or the pump 405.

[0068] The pump 405 (separating liquid introducing means) introduces the separating liquid 202 from the second opening 107 through the solid phase 106 into the flow channel 104 .

[0069] The vacuum pump 403 is a pump capable of reducing the pressure from atmospheric pressure to, for example, about 5 kPa to 0.01 kPa. A higher vacuum level allows more air to be released from the microchamber 105, allowing more sample solution 201 to be added.

[0070] In addition, the pump 405 for introducing the separation liquid 202 uses, for example, a syringe pump. The syringe pump pushes out the pre-set separation liquid 202 and introduces the separation liquid 202 into the flow path 104. It should be noted that the pump 405 can also suck the separation liquid 202 in a pre-set container and push the sucked separation liquid 202 into the flow path 104. The pump 405 can also pressurize the separation liquid 202 to introduce it into the flow path, and can be, for example, a quantitative ejection pump, a diaphragm pump, a rotary pump, etc. By pressurizing the separation liquid 202 when introducing it into the flow path 104, the separation liquid 202 can be pressed in even in the presence of resistance from the solid phase 106 or flow path resistance.

[0071] Specifically, the solid phase 106 uses a porous membrane. For example, the solid phase 106 uses PTFE (polytetrafluoroethylene). Since the solid phase 106 is porous and breathable, it can allow air to pass through. Since it has water repellency, it can stop the sample solution 201. Since it has lipophilicity, it can pass the separation liquid 202. In addition, the solid phase 106 can also use PFA (perfluoroalkoxyalkylene), FEP (fluoroethylene propylene), ETFE (ethylene tetrafluoroethylene), etc., fluorine-based water-repellent membranes, silicon-based water-repellent membranes, polymer-based water-repellent membranes, nanoporous membranes. In addition, the solid phase 106 may not be a membrane, but may be a structure in which hydrophobic beads are filled in the flow path 104. Since the beads are hydrophobic, they can be water-repellent for the sample solution 201 and stop it. As a porous membrane, a hydrophobic membrane having a contact angle with water, for example, more than 80 degrees can be used. In addition, the solid phase 106 is made of a lipophilic material that allows the separation liquid 202 to penetrate and pass through, so that the separation liquid 202 can pass through.

[0072] In the past, for example, as in the method disclosed in non-patent document 1, a valve was used instead of the above-mentioned solid phase 106. In non-patent document 1, when the valve is closed, the sample solution can be stopped, and when the valve is opened, air can be discharged and a separation liquid can be introduced. However, in the case of a pinch valve such as a tube valve that squeezes the tube, a portion from the flow path to the tube and a volume for the sample solution to pass through the tube are required. The sample solution remaining in the tube cannot be used for measurement and thus becomes a dead volume. Due to this dead volume, the measurable target DNA is reduced, resulting in a decrease in detection sensitivity. By using the solid phase 106 of Example 1, the solid phase 106 can be arranged near the microchamber 105, which can reduce the dead volume of the sample solution 201.

[0073] As described above, by using the solid phase 106 of Example 1, the solid phase 106 can be attached to the flow path 104, thereby reducing dead volume such as a valve. Although the sample solution 201 remaining in the flow path 104 may become a dead volume, by designing and processing the flow path 104 to have a smaller cross-sectional area, the volume of the sample solution 201 remaining in the flow path 104 can be minimized, thereby reducing the dead volume of the sample solution 201.

[0074] In addition, in Example 1, valve 103 is used before sample solution 201 is introduced into flow path 104. Valve 103 is closed when vacuuming and opened when sample solution 201 is introduced. Valve 103 can also be a disposable valve that is opened only once when sample solution 201 is introduced into flow path 104. Specifically, for example, a resin film is used as a valve, which is normally closed and then opened when sample solution 201 is introduced by destroying the film. Thus, a low-cost and compact device can be provided. It should be noted that a solenoid valve or a pinch valve of a squeeze tube can also be used as valve 103.

[0075] As the separation liquid 202, a liquid having a physical property of not being mixed with the sample solution 201 is used. Specifically, for example, silicone oil, mineral oil, or other oil, paraffin, or a photocurable resin can be used as the separation liquid 202.

[0076] The separation liquid 202 can also be a photocurable resin. When using a photocurable resin to separate the sample solution 201, the photocurable resin is introduced into the flow path 104 in a liquid state. Thereafter, the photocurable resin is cured by irradiation with light such as ultraviolet light, and can be separated and sealed in the multiple microchambers 105 containing the sample solution 201. In addition, the separation liquid can also be not one, but multiple. Specifically, for example, photocurable resin and oil are introduced in sequence from the second opening 107. The photocurable resin reaches the vicinity of the valve 103, and the flow path 104 is filled with oil. Thereafter, by curing the photocurable resin, it is possible to configure the end of the flow path 104 to be sealed with the photocurable resin and the microchambers 105 to be separated with oil.

[0077] The sample solution 201 is a solution containing, for example, a DNA to be measured, a polymerase, a buffer, primers, and a probe, and its volume is adjusted to about 10 μL to about 50 μL.

[0078] In Example 1, compared to a conventional structure using valves on both sides of the flow channel, the dead volume of the sample solution 201 can be reduced. Therefore, most of the adjusted sample solution 201 can be used for measurement, enabling highly sensitive detection.

[0079] It should be noted that when the sample solution 201 is introduced into the first opening 102, another liquid (separation liquid) such as oil can also be continuously added with the sample solution 201. In this way, the sample solution 201 can be pushed into the microchamber 105 by the oil. In this way, the flow path 104 can be filled with the separation liquid such as oil, thereby further reducing the dead volume of the sample solution 201 in the flow path 104. In addition, when the separation liquid 202 is introduced, the liquid pushed back to the first opening 102 becomes the oil (separation liquid), which acts as a cover, thereby preventing the nucleic acid in the sample solution 201 from diffusing into the atmosphere.

[0080] In this manner, the sample solution separation device 101, in which the sample solution 201 is separated and introduced into the plurality of microchambers 105, performs PCR thermal cycling. Through thermal cycling, the target DNA is amplified by PCR within the plurality of microchambers 105. The thermal cycling is set, for example, at a high temperature of 95°C for 20 seconds and a low temperature of 60°C for 40 seconds. The high temperature dissociates the double strands, while the low temperature allows annealing and extension, thereby performing PCR amplification. Fluorescence measurement of the amplified products allows detection of the target DNA separated and introduced into the plurality of microchambers 105.

[0081] The volume of the microchamber 105 is, for example, several picoliters to several nanoliters, and the number of microchambers 105 is approximately several thousand to several million. In digital PCR, by dividing the target DNA into multiple microchambers 105, the amount of background DNA can be reduced, allowing for highly sensitive detection of the target DNA.

[0082] It should be noted that, in this embodiment, the movement mechanism 406 (switching unit) is used to switch between the opening contact portion 402 and the opening contact portion 404 , but the flow path may be switched by a solenoid valve or the like.

[0083] The microchamber 105 is connected to the flow path 104 by a thin connection flow path that branches from the main flow path. Through this thin connection flow path, multiple microchambers 105 are steadily separated. When the separation liquid 202 is introduced into the flow path 104, the sample solution 201 introduced into the microchamber 105 flows in the main flow path. The flow path cross-sectional area of ​​the connection flow path is smaller than that of the main flow path. In addition, since the microchamber 105 at the front end of the connection flow path is filled with sample solution 201, the separation liquid 202 does not enter the microchamber 105 from the connection flow path, but flows in the main flow path. In addition, since interfacial tension acts between the sample solution 201 and the separation liquid 202, the sample solution 201 becomes droplet-shaped in the microchamber 105, and the separation liquid 202 separates multiple microchambers 105. By connecting the connecting flow channels having a smaller cross-sectional area than the main flow channel to the plurality of microchambers 105 , the separation solution 202 does not enter the microchambers 105 but passes through the flow channels 104 , thereby separating the sample solution 201 .

[0084] (Example 2)

[0085] In Example 1, an example in which the solid phase 106 is arranged at one location in the flow channel 104 has been described, but the present invention is not limited thereto. Figure 5It is a structural diagram of the sample solution separation device 501 of Example 2. The sample solution separation device 501 includes a first opening 102, a valve 103, a flow path 104, a plurality of microchambers 105, a plurality of solid phases 506a to 506d, and a second opening 107. It should be noted that the same descriptions as in Example 1 are appropriately omitted. The flow path 104 branches in order to connect with the plurality of microchambers 105. After the plurality of branch flow paths 104a to 104d branching from the main flow path are connected to the plurality of microchambers 105, they merge again and are connected to the second opening 107. The solid phases 501a to 501d that allow air to pass, stop the sample solution 201, and allow the separation liquid 202 to pass can be set at a plurality of locations between the microchamber 105 and the second opening 107. In Figure 5 In the example, solid phases 501a to 501d are respectively provided in the plurality of branch flow paths 104a to 104d of the flow path 104. The number of the plurality of branch flow paths 104a to 104d and the number of solid phases 501a to 501d are not limited to 4, and may be 2 to 3, or may be 5 or more.

[0086] As in Example 2, by arranging the solid phases 501 a to 501 d closer to the microchamber 105 than in Example 1, the sample solution 201 remaining in the flow channel 104 can be reduced, and the dead volume of the sample solution 201 can be reduced.

[0087] (Example 3)

[0088] use Figure 6A 、 Figure 6B and Figure 7 The sample solution separation device 601 of Example 3 will be described. Figure 6A is a top view of the sample solution separation device 601 of Example 3, Figure 6B 3 is a side view of the sample solution separation device 601 of Example 3. Figure 6A and Figure 6B As shown, the sample solution separation device 601 includes a first opening 602, a valve 603, a flow path 604, a plurality of microchambers 605, a solid phase 606, and a second opening 607. It should be noted that the same descriptions as in Example 1 and Example 2 are appropriately omitted. The flow path 604 and the microchamber 605 are processed on a substrate 608. In addition, through holes 610 and 611 that penetrate the substrate 608 are formed at one end and the other end of the flow path 604. The through hole 610 is connected to the first opening 602 via the valve 603, and the through hole 611 is connected to the second opening 607 via the solid phase 606. In addition, the side of the substrate 608 on which the plurality of microchambers 605 and the flow path 604 are processed is sealed by a membrane 609.

[0089] Specifically, the flow path 604 of the sample solution separation device 601 of Example 3 includes a horizontal flow path 604a extending horizontally and connected to the plurality of microchambers 605, and a vertical flow path 604b (through holes 610 and 611) extending vertically and connected to the horizontal flow path 604a. Furthermore, the solid phase 606 is provided at the upper end of the vertical flow path 604b (through hole 611).

[0090] The substrate 608 or film 609 may be made of a resin, but the present invention is not limited thereto. For example, the substrate 608 or film 609 may be made of a COP (cyclic olefin polymer) or a COC (cyclic olefin copolymer) material, which exhibits low autofluorescence. Alternatively, the substrate 608 or film 609 may be made of polycarbonate, polypropylene, PMMA (methacrylic resin), or the like. Furthermore, a portion of the substrate 608 or film 609 may be made of a metal such as aluminum, which has high thermal conductivity, or carbon, which suppresses light reflection.

[0091] The valve 603 is a normally closed seal configured to block the through hole 610. It is a disposable valve that changes from a closed state to an open state by opening a hole. A first opening 602 having a volume capable of receiving the sample solution 201 is configured on the upper portion of the valve 603. In addition, the solid phase 606 is film-shaped and configured to block the through hole 611. A second opening 607 having a volume capable of receiving the separation liquid 202 is configured on the upper portion of the solid phase 606. By setting it to this structure, the valve 603 and the solid phase 606 can be easily configured on the substrate 608. In addition, since the solid phase 606 can be configured near the flow path 604, the dead volume of the sample solution 291 can be reduced.

[0092] Figure 7 This is a structural diagram of a sample solution separation system 700 that separates a sample solution 201 and introduces the separated sample solution into the sample solution separation device 601 of Example 3. The sample solution separation system 700 includes a vacuum pump 701, a pressure sensor 702, a filter 703, a solenoid valve 704, a moving mechanism 705, an opening contact portion 706, an opening contact portion 707, a solenoid valve 708, a liquid pump 709, a container 710, a filter 711, a pressure sensor 712, a pressurizing pump 713, a valve control mechanism 714, an opening cover 715, a light source 717, a thermostat 718, a controller 719, and the sample solution separation device 601.

[0093] The sample solution 201 is pre-filled in the first opening 602 and placed in contact with the upper portion of the valve 603. At this time, the valve 603 is closed. An opening cover 715 is provided on the first opening 602. A pointed tip 716, capable of breaking the seal (valve 603), is provided on the inner side of the upper surface of the opening cover 715. The upper surface of the opening cover 715 is formed of a stretchable material, such as an elastomer or rubber.

[0094] The second opening 607 of the sample solution separation device 601 is connected to the opening contact portion 706 by controlling the movement mechanism 705 (switching unit). Furthermore, the solenoid valve 704 connects or disconnects the vacuum pump 701 from the opening contact portion 706. The vacuum pump 701 evacuates the flow path 604 and the multiple microchambers 605 of the sample solution separation device 601. After the flow path 604 and the multiple microchambers 605 are sufficiently depressurized, the valve control mechanism 714 (valve control unit) presses the top of the opening cover 715, causing the pointed member 716 to break the valve 603. This opens the valve 603. As mentioned above, the valve 603 is formed of a sealing member made of a thin film such as aluminum or resin, which can be broken by the pointed member 716. Since the flow path 604 and the multiple microchambers 605 are depressurized, the sample solution 201 is introduced into the flow path 604 and the multiple microchambers 605, filling the flow path 604 and the multiple microchambers 605 with the sample solution 201. Since the solid phase 606 is water-repellent, the sample solution 201 passing through the flow path 604 stops at the solid phase 606. In addition, since the solid phase 606 is air-permeable, the flow path 604 and the microchamber 605 can be negatively pressurized by the vacuum pump 701 until the tip of the sample solution 201 reaches the solid phase 606, thereby increasing the degree of vacuum.

[0095] The electromagnetic valve 704 is used to seal the flow path 604, connect the vacuum pump 701 to the opening contact portion 706, and open it to the atmosphere. By opening the second opening 607 to the atmosphere, the flow path 604 of the sample solution separation device 601 is opened to atmospheric pressure. Thereafter, the moving mechanism 705 connects the opening contact portion 707 to the second opening 607. The electromagnetic valve 708 connects the opening contact portion 707 to the liquid pump 709. The liquid pump 709 draws the separation liquid 202 pre-set in the container 710 and discharges it toward the opening contact portion 707. The separation liquid 202 is introduced into the flow path 604 through the second opening 607 and the solid phase 606.

[0096] In addition, the solenoid valve 708 switches its connection destination to the second opening 607 from the liquid pump 709 to the pressure pump 713. The pressure pump 713 pressurizes the separation liquid 202 and forces it into the flow path 604. The pressure pump 713 applies pressure to a pressure that exceeds the flow resistance of the flow path 604 and the resistance when passing through the solid phase 606, thereby introducing the separation liquid 202 into the flow path 604. The pressure during pressurization is determined by the separation liquid 202, the dimensions of the flow path 604, the properties of the solid phase 606, and the like, and is, for example, approximately 10 kPa to 200 kPa. The sample solution 201 in the flow path 604 is pushed back toward the first opening 602, and the flow path 604 is filled with the separation liquid 202. In this way, the sample solution 201 remains in each microchamber 605 and is separated by the separation liquid 202. Furthermore, by introducing the separation liquid 202 into the flow path 604 under pressure, the separation liquid 202 can be introduced into the flow path 604 while suppressing the expansion of bubbles that remain for some reason.

[0097] Alternatively, the separation liquid 202 may be a photocurable resin. By using a photocurable resin that does not mix with the sample solution 201, the sample solution 201 can be separated into each microchamber 605. In this case, after the liquid photocurable resin is introduced into the flow path 604, the light source 717 is used to cure the photocurable resin. This allows the sample solution 201 to be sealed within the microchamber 605.

[0098] In addition, when introducing the sample solution 201 and the separating liquid 202, the temperature of the sample solution separation device 601 can also be controlled using a thermostat 718. Generally speaking, the viscosity of the sample solution 201 and the separating liquid 202 decreases at high temperatures. Therefore, by heating the sample solution 201 and the separating liquid 202 compared to room temperature, the viscosity of the sample solution 201 and the separating liquid 202 decreases, making it easier for the sample solution 201 and the separating liquid 202 to enter the flow path 604 and the microchamber 605. As a result, the introduction time of the solutions (sample solution 201 and the separating liquid 202) can be shortened, and the sample solution 201 can be steadily added to all the microchambers 605. The thermostat 718 is preferably controlled to heat the sample solution to approximately 35°C to 70°C, for example.

[0099] The pressure during evacuation by the vacuum pump 701 is monitored and controlled by the pressure sensor 702. The pressure during pressurization by the pressure pump 713 is monitored and controlled by the pressure sensor 712. Filters 703 and 711 prevent impurities from entering the sample solution 201 and the separation liquid 202.

[0100] The control of the above series of operations is performed by the controller 719. The controller 719 includes a processor 720, a memory 721, and an interface 722. The processor 720 executes various programs through the memory 721 and outputs control signals for controlling the above operations via the interface 722.

[0101] The valve control mechanism 714 is, for example, a solenoid, and moves when current flows to open the valve 603. The moving mechanism 705 is, for example, a two-axis motor drive mechanism in the horizontal and vertical directions.

[0102] By using a solid phase 606 having gas permeability, hydrophobicity, and lipophilicity, it is possible to control the evacuation of the flow path 604, the stopping of the sample solution 201, and the passage of the separation liquid 202 without using valves. In addition, the solid phase 606 can be arranged near the microchamber 605, thereby preventing waste of the sample solution 201 and allowing the sample solution 201 to be divided and placed in the microchamber 605.

[0103] In addition, as valve 603, by using the disposable valve of the structure of perforation on film, the valve of simple structure can be installed. Because sample solution 201 contacts with valve 603, therefore in order to prevent carry-over, need disposable use, so disposable valve is useful. In addition, valve 603 is a valve with simple structure, therefore valve can be provided at low cost. In addition, using light-curing resin as separating liquid 202, after being put into flow path 604 in liquid form, by solidification, microchamber 605 can be sealed. Thus, it is no longer necessary to close the first opening portion 602 and the second opening portion 607, and the structure can be simplified.

[0104] It should be noted that the valve 603 can also be opened with a pipette tip. The sample solution separation device 601 of Example 3 can also be applied to an automated system connected to a sample pre-treatment such as nucleic acid extraction, reagent mixing, and injection. For example, the sample collected in the blood collection tube is centrifuged to remove the plasma, and the nucleic acid in the plasma is extracted by refining the plasma. The nucleic acid is mixed with the reagent, and the target nucleic acid is detected by digital PCR. At this time, a dispensing system that attracts and ejects the mixed sample solution through a pipette tip is used to inject the sample solution 201 into the first opening 602. At this time, the front end of the pipette tip can also be used to destroy the valve and introduce the sample solution 201 into the flow path.

[0105] In addition, by continuously introducing oil (separating liquid) from the first opening 602 and the sample solution 201, the dead volume of the sample solution 201 can be further reduced. Specifically, after the sample solution 201 is introduced into the microchamber 605 by negative pressure, the oil (separating liquid) continuously enters the flow path 604, thereby reducing the sample solution 201 remaining in the flow path 604. By adding the separating liquid 202 from the second opening 607 in the direction opposite to the sample solution 201 in this state, the sample solution 201 in the flow path 604 can be completely replaced with the separating liquid 202, separating the multiple microchambers 605. In addition, the oil (separating liquid) continuously introduced with the sample solution 201 has the function of a cover to prevent the nucleic acid in the sample solution 201 from becoming an aerosol and being released into the atmosphere. Thus, compared with the case where only the sample solution 201 is introduced from the first opening 602, the dead volume of the sample solution 201 can be further reduced.

[0106] It should be noted that in this embodiment, the separation liquid 202 in the container 710 is introduced into the flow path 604 through the second opening 607, but the present invention is not limited thereto. For example, the separation liquid 202 may be pre-filled in the sample solution separation device 601, and the separation liquid 202 filled in the sample solution separation device 601 may be introduced into the flow path 604.

[0107] (Example 4)

[0108] In the third embodiment described above, the thin film solid phase 606 is provided on the upper end portion of the through hole 611 of the substrate 608 , but the present invention is not limited thereto. Figure 8 This is a structural diagram of the sample solution separation device 801 of Example 4. The solid phase 806 of the sample solution separation device 801 of Example 4 is a plurality of microparticles having air permeability, hydrophobicity, and lipophilicity. A plurality of microparticles are stuffed into the through hole 611 (vertical flow path) to form the solid phase 806. The microparticles are, for example, beads having a size of 0.1 μm to 10 μm. The microparticles are easier to introduce into the flow path 604 than the membrane. In addition, since air can pass between the microparticles, vacuuming can be performed. In addition, since the microparticles are water-repellent, the sample solution 201 can be stopped. In addition, since the microparticles are lipophilic, they can pass through the separation liquid 202.

[0109] In addition, in Example 3, valve 603 is a disposable valve, but the present invention is not limited to this. For example, valve 803 of the sample solution separation device 800 of Example 4 can be a retractable tube. The tube extends to connect to the through hole 610 of the substrate 608, forming valve 803. For example, valve 803 is a silicone tube. The silicone tube is compressed to form a pinch valve that stops the flow in the tube. By using a pinch valve, it can be opened and closed multiple times. Valve 803 is closed when vacuuming and opened when the sample solution 201 is added. In addition, after the separation liquid 202 is introduced, valve 803 is closed to seal the sample solution 201 and the separation liquid 202.

[0110] (Example 5)

[0111] The sample solution separation device of the present invention can be used for digital PCR. Figure 9 This is a block diagram of a digital PCR system 900 according to Example 5. The digital PCR system 900 (sample solution separation system) includes a sample solution separation device 601, a valve control mechanism 901, a pump 902, a liquid pump 903, a thermostat 913, an optical system 914 (measurement unit), and an analysis unit 912. The optical system 914 includes a light source 904, a lens 905, a lens 908, a lens 910, a bandpass filter 906, a bandpass filter 909, a dichroic mirror 907, and a CMOS sensor 911.

[0112] The sample solution 201 is introduced into the sample solution separation device 601 through the first opening 602. The pump 902 is driven to deaerate the air within the flow path 604 and microchamber 605 through the second opening 607, creating a negative pressure. The valve control mechanism 901 opens the valve 603 to introduce the sample solution 201 into the flow path 604 and microchamber 605. Subsequently, the liquid pump 903 is driven to introduce the separation liquid 202 into the flow path 604 through the second opening 607. This allows the sample solution 201 to be separated and introduced into the multiple microchambers 605.

[0113] Here, sample solution 201 is a PCR reaction solution containing the target DNA, polymerase, buffer, primers, and probes. A thermal cycling process is performed on a device that separates sample solution 201, which serves as the PCR reaction solution, and places it into multiple microchambers 605. During the thermal cycling process, thermostat 913 is controlled to a temperature range for denaturation, annealing, and expansion. Thermostat 913 may be composed of a heater, Peltier element, or the like, but the present invention is not limited thereto. Through the thermal cycling process, the target DNA is amplified while it is present within microchambers 605.

[0114] Optical system 914 (measurement unit) measures the target nucleic acid within sample solution 201 that has undergone PCR amplification. Light emitted from light source 904 is collimated by lens 905, passes through bandpass filter 906 to allow light of a specified wavelength to pass through, is reflected by dichroic mirror 907, and then passes through lens 908 to illuminate sample solution separation device 601. Fluorescence from microchambers 605 of sample solution separation device 601 passes through lens 908, dichroic mirror 907, bandpass filter 909, and lens 910, and is imaged by CMOS sensor 911. Analysis unit 912 performs analysis based on the image formed by CMOS sensor 911, detecting the target DNA within each microchamber.

[0115] Figure 10 This is an operation flow chart of the digital PCR system 900 of Example 5.

[0116] The valve control mechanism 901 opens the valve 603 to introduce the sample solution 201 provided in the first opening 602 into the sample solution separation device 601 ( S1001 ).

[0117] The liquid pump 903 introduces the separation liquid 202 into the sample solution separation device 601 from the second opening 607 ( S1002 ).

[0118] Next, the thermostat 913 causes the sample solution separation device 601 to perform thermal cycling ( S1003 ).

[0119] The optical system 914 irradiates light to the sample solution separation device 601 and measures the fluorescence intensity of each microchamber 605 ( S1004 ).

[0120] Then, the analysis unit 915 analyzes the fluorescence intensity measured by the optical system 914 and detects the target DNA in the sample solution 201 ( S1005 ).

[0121] It should be noted that the sample solution 201 and the separating liquid 202 may be heated during their introduction. This changes the viscosity of the sample solution 201 and the separating liquid 202, facilitating their introduction into the flow path 604. Furthermore, the time required to introduce the sample solution 201 and the separating liquid 202 can be shortened. The thermostat for heating the sample solution 201 and the separating liquid 202 may be the thermostat 913 that performs the thermal cycle, or a thermostat separate from the thermostat 913.

[0122] Asymmetric PCR can also be performed by varying the concentrations of the forward and reverse primers in the PCR reaction solution, thereby increasing the amplification of a single strand of single-stranded DNA within the amplified product. This allows for detection of single-stranded DNA using molecular beacons. Furthermore, during fluorescence measurement, the temperature of the sample solution separation device is controlled to perform melting curve analysis. Fluorescence measurement uses multiple filters, allowing for multi-color measurement. By using the color, intensity, and melting temperature of the fluorescence from each microchamber to identify the target DNA, highly multiplexed and sensitive measurements are possible.

[0123] (Sample solution separation method)

[0124] Here, a method for separating the sample solution 201 into the plurality of microchambers 105 will be described. The sample solution separation method includes the steps of preparing the sample solution separation device 101 described above, degassing the air within the plurality of microchambers 105 and the flow path 104 via the solid phase 106, opening the valve 103 to introduce the sample solution 201 into the plurality of microchambers 105 and the flow path 104 through the first opening 102, and introducing the separation solution 202 into the flow path 104 through the second opening 107 via the solid phase 106.

[0125] It should be noted that the prepared sample solution separation device 101 may also be the sample solution separation device 501 , 601 or 801 .

[0126] In addition, the sample solution separation method also includes a process of performing PCR (polymerase chain reaction) thermal cycling on the sample solution separation device 101 that separates the sample solution 201 in multiple microchambers 105, a process of measuring the fluorescence intensity of the multiple microchambers 105, and a process of analyzing the fluorescence intensity to detect the target DNA in the sample solution 201.

[0127] Note that the process of introducing the sample solution 201 into the plurality of microchambers 105 and the flow path 104 from the first opening 102 includes the process of introducing the sample solution 201 and the oil (separation liquid) continuous with the sample solution 201 from the first opening 102 .

[0128] (Variation)

[0129] The present disclosure is not limited to the above-described embodiments, but includes various variations. For example, the above-described embodiments are described in detail to facilitate understanding of the present disclosure and are not necessarily limited to having all the structures described. In addition, a portion of the structure of a certain embodiment can be replaced with the structure of another embodiment, and the structure of another embodiment can be added to the structure of a certain embodiment. In addition, with respect to a portion of the structure of each embodiment, other structures can be added, deleted, or replaced.

[0130] For example, in the above-described embodiment, a separation liquid is used to separate the plurality of microchambers 105 , but the present invention is not limited thereto and a separation gas may also be used. In this case, the solid phase only needs to have physical properties that allow the separation gas to pass through.

[0131] Description of Reference Numerals

[0132] 101, 501, 601, 801: sample solution separation device,

[0133] 102: first opening,

[0134] 103: valve,

[0135] 104: Flow path,

[0136] 105: Micro chamber,

[0137] 106: solid phase,

[0138] 107: second opening,

[0139] 201: sample solution,

[0140] 202: separation liquid,

[0141] 400, 700: sample solution separation system,

[0142] 401: Valve control mechanism,

[0143] 402, 404: opening contact portion,

[0144] 403, 405: pump,

[0145] 406: Mobile mechanism,

[0146] 506a, 506b, 506c, 506d: solid phase,

[0147] 602: first opening,

[0148] 603: valve,

[0149] 604: Flow path,

[0150] 604a: Horizontal flow path,

[0151] 604b: Vertical flow path,

[0152] 605: Micro chamber,

[0153] 606: solid phase,

[0154] 607: second opening,

[0155] 608: Substrate,

[0156] 609: membrane,

[0157] 610, 611: through holes,

[0158] 701: vacuum pump,

[0159] 702, 712: pressure sensor,

[0160] 703, 711: filter,

[0161] 704, 708: solenoid valve,

[0162] 705: mobile mechanism,

[0163] 706, 707: opening contact portion,

[0164] 709: Liquid pump,

[0165] 710: container,

[0166] 713: pressure pump,

[0167] 714: Valve control mechanism,

[0168] 715: Cover for opening,

[0169] 716: pointed parts,

[0170] 717: Light source,

[0171] 718: Thermostat,

[0172] 719: Controller,

[0173] 720: processor,

[0174] 721: Memory,

[0175] 722: interface,

[0176] 803: valve,

[0177] 806: solid phase,

[0178] 900: Digital PCR system,

[0179] 901: Valve control mechanism,

[0180] 902: Pump,

[0181] 903: Liquid pump,

[0182] 904: Light source,

[0183] 905, 908, 910: lenses,

[0184] 906, 909: Bandpass filter,

[0185] 907: dichroic mirror,

[0186] 911: CMOS sensor,

[0187] 912: Analysis Department,

[0188] 913: Thermostat,

[0189] 914: Optical system.

Claims

1. A sample solution separation device for separating a sample solution, characterized in that: have: Multiple microchambers, a flow path connecting the plurality of microchambers, a first opening serving as an inlet for introducing the sample solution into the flow path; a valve disposed between the first opening and the plurality of microchambers, a second opening serving as an inlet for a separation liquid to enter the flow path and provided on the opposite side of the first opening across the plurality of microchambers, wherein the separation liquid separates the plurality of microchambers containing the sample solution; and a solid phase disposed between the second opening and the plurality of microchambers; The solid phase has air permeability, is water-repellent to the sample solution, and is permeable to the separation solution.

2. The sample solution separation device according to claim 1, characterized in that: The flow path has a plurality of branch flow paths respectively connected to the microchambers. The solid phase is respectively provided in the plurality of branch flow channels.

3. The sample solution separation device according to claim 1, characterized in that: The flow path has: a horizontal flow path connected to the plurality of microchambers and extending in a horizontal direction, and a vertical flow path connected to the horizontal flow path and extending in the vertical direction; The solid phase is provided at the upper end of the vertical flow path.

4. The sample solution separation device according to claim 1, characterized in that: The flow path has: a horizontal flow path connected to the plurality of microchambers and extending in a horizontal direction, and a vertical flow path connected to the horizontal flow path and extending in the vertical direction; The solid phase is a plurality of microparticles disposed in the vertical flow path.

5. The sample solution separation device according to claim 1, characterized in that: The valve is a disposable valve that is opened only once when the sample solution is introduced into the flow path.

6. The sample solution separation device according to claim 1, characterized in that: The solid phase is a porous membrane that is water-repellent to the sample solution and permeable to the separation solution.

7. The sample solution separation device according to claim 6, characterized in that: The material of the porous membrane is PTFE, i.e. polytetrafluoroethylene.

8. The sample solution separation device according to claim 1, characterized in that: The separation liquid is a photocurable resin, and the separation liquid is cured by light irradiation in the flow channel.

9. The sample solution separation device according to claim 1, characterized in that: The separation liquid is oil having a physical property of not being mixed with the sample solution.

10. A sample solution separation system, characterized in that: It has a sample solution separation device, a pump, a valve control unit and a separation liquid introduction unit; The sample solution separation device separates the sample solution, and comprises: Multiple microchambers, a flow path connecting the plurality of microchambers, a first opening serving as an inlet for introducing the sample solution into the flow path; a valve disposed between the first opening and the plurality of microchambers, a second opening serving as an inlet for a separation liquid to enter the flow path and provided on the opposite side of the first opening across the plurality of microchambers, wherein the separation liquid separates the plurality of microchambers containing the sample solution; and a solid phase disposed between the second opening and the plurality of microchambers; The solid phase has air permeability, is water-repellent to the sample solution, and is permeable to the separation liquid; The pump degases the air in the plurality of microchambers and the flow path from the second opening through the solid phase; The valve control unit causes the valve to open; The separation liquid introduction unit introduces the separation liquid from the second opening.

11. The sample solution separation system according to claim 10, characterized in that: The device further includes a switching unit that switches the connection destination of the second opening to the pump or the separation liquid introduction unit.

12. The sample solution separation system according to claim 10, characterized in that: The device further includes a pressure pump for pressurizing the separation liquid and introducing the pressurized separation liquid into the flow path.

13. The sample solution separation system according to claim 10, characterized in that: The device further comprises a measuring unit for measuring the target nucleic acid in the sample solution subjected to PCR (polymerase chain reaction) in the microchamber.

14. A method for separating a sample solution, characterized in that: The method includes preparing a sample solution separation device for separating a sample solution, wherein the sample solution separation device comprises: Multiple microchambers, a flow path connecting the plurality of microchambers, a first opening serving as an inlet for introducing the sample solution into the flow path; a valve disposed between the first opening and the plurality of microchambers, a second opening serving as an inlet for a separation liquid to enter the flow path and provided on the opposite side of the first opening across the plurality of microchambers, wherein the separation liquid separates the plurality of microchambers containing the sample solution; and a solid phase disposed between the second opening and the plurality of microchambers, the solid phase having air permeability, water repellency to the sample solution, and permeability to the separation liquid; The sample solution separation method further comprises: The process of degassing the air in the plurality of microchambers and the flow path through the solid phase, a process of opening the valve and introducing the sample solution into the plurality of microchambers and the flow path from the first opening, and A process of introducing the separation liquid into the flow channel from the second opening via the solid phase.

15. The sample solution separation method according to claim 14, characterized in that: Also features: The sample solution separation device that separates the sample solution in the plurality of microchambers performs a thermal cycle of PCR (polymerase chain reaction), a process of measuring the fluorescence intensity of the plurality of microchambers, and The fluorescence intensity is analyzed to detect the target DNA in the sample solution.

16. The sample solution separation method according to claim 14, characterized in that: The process of introducing the sample solution into the plurality of microchambers and the flow channel from the first opening includes the process of introducing the sample solution and a separation solution continuous with the sample solution from the first opening.

Images