Particle separation device

Abstract

To provide particle separators that enable processing of a large amount of sample liquid in a short time and at high speed with a simple configuration.SOLUTION: A particle separator 10 comprises a first separation unit 20 provided with a DLD microchannel for flowing a sample liquid Sa containing a target particle, and a second separation unit 30 provided with a DLD microchannel for flowing the target particle concentrate Sb discharged from the first separation unit 20. The first separation unit 20 comprises: a sample liquid introduction port 21 into which the sample liquid is introduced, and a DLD flow path unit that has a DLD microchannel structure and flows and separates the sample liquid Sa introduced from the sample liquid introduction port 21 into a liquid in which the concentration of the target particles has increased and a liquid in which the target particles have been almost removed; a target particle concentrate discharge port 22 that discharges a liquid having an increased concentration of target particles as a target particle concentrate Sb; and a target particle removal liquid discharge port 23 that discharges the liquid from which the target particles have been almost removed.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a particle separation device that separates target particles from a liquid containing the target particles. [Background technology]

[0002] It is known that tumor cells are released into the bloodstream from primary tumor tissues or metastatic tumor tissues and circulate in the body with the blood. Such tumor cells are called circulating tumor cells (CTCs), and it is expected that isolating and examining circulating tumor cells from peripheral blood will provide information for understanding the progression of cancer and the effectiveness of treatment, as well as for early detection of recurrence and metastasis.

[0003] As a particle separation technique for easily separating circulating tumor cells in the blood, a method using the deterministic lateral displacement (DLD) method, which is a type of hydrodynamic laminar flow separation, is known, taking advantage of the fact that circulating tumor cells in the blood tend to be larger in size than blood cells (see, for example, non-patent document 1).

[0004] As an example of a separation device that utilizes this separation technology, the following Patent Document 1 discloses a particle separation device that can introduce equal amounts of sample liquid and buffer liquid into a DLD microchannel chip by applying pressure from a pressure supply unit through a branch pipe to push out the sample liquid and buffer liquid.

[0005] Furthermore, for example, Patent Document 2 listed below discloses various design aspects of a chip (device) configured by multiplexing an array having one or more stages in order to concentrate or separate particles in a sample.

[0006] Furthermore, for example, the following non-patent document 2 discloses a configuration in which a first chip is equipped with a DLD microchannel into which a buffer solution and a sample solution (blood) are introduced and which separates the buffer solution containing CTCs and white blood cells from a solution containing red blood cells, platelets, etc., and a second chip is equipped with a channel for inertial force focusing and a separation unit using magnetic migration and which separates the buffer solution containing CTCs and white blood cells into a solution containing CTCs and a solution containing white blood cells, connected in series. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] International Publication No. 2019 / 172428 [Patent Document 2] Special Publication No. 2008-538283 [Non-patent literature]

[0008] [Non-Patent Document 1] Huang et al.,"Continuous Particle Separation Through Deterministic Lateral Displacement",Science Vol.304,pp.987-990,2004 [Non-Patent Document 2] Karabacak, et al., "Microfluidic, marker-free isolation of circulating tumor cells from blood samples", Nature Protocols Vol.9, pp.694-710,2014 Summary of the Invention [Problem to be solved by the invention]

[0009] However, the particle separation device disclosed in Patent Document 1 has a problem in that the processing speed of the DLD microchannel chip is limited and it takes a huge amount of time to process a large amount of sample liquid, making it practically difficult to separate target particles from a large amount of sample liquid.

[0010] In addition, the chip disclosed in Patent Document 2 is complex, and it is not easy to manufacture a chip equipped with such a complex DLD microchannel. In addition, there is a problem that there is a limit to the amount of sample that can be processed by a single chip.

[0011] Furthermore, with regard to the configuration disclosed in Non-Patent Document 2, like the particle separation device disclosed in Patent Document 1, there is a problem in that the processing speed of the DLD microchannel chip is limited (in the configuration disclosed in Non-Patent Document 2, the processing speed of sample liquid is approximately 120 μL per minute), and it takes a huge amount of time to process a large amount of sample liquid, making it practically difficult to separate target particles from a large amount of sample liquid.

[0012] The present invention has been made in consideration of these points, and has an object to provide a particle separation device that has a simple configuration and is capable of processing a large amount of sample liquid at high speed in a short period of time. [Means for solving the problem]

[0013] In order to achieve the above object, a particle separator according to the present invention comprises: A first separation unit including a DLD microchannel in which a plurality of fine pillars are arranged to allow a sample liquid containing target particles to flow; a second separation section connected to the first separation section and including a DLD microchannel having a plurality of fine pillars arranged therein for circulating the target particle concentrate discharged from the first separation section; 1. A particle separator comprising: The first separation unit is a sample liquid inlet into which the sample liquid is introduced; a DLD flow path section that includes a DLD microchannel structure and that circulates the sample liquid introduced from the sample liquid inlet port to separate the sample liquid into a liquid with an increased concentration of the target particles and a liquid from which the target particles have been almost completely removed; a target particle concentrated liquid outlet that discharges the liquid in which the concentration of the target particles separated in the DLD flow path portion has been increased as the target particle concentrated liquid; a target particle removed liquid outlet for discharging a liquid from which the target particles separated in the DLD flow path portion have been almost completely removed; The present invention is characterized by comprising:

[0014] According to the above configuration, the first separation section separates the sample liquid into a liquid with an increased concentration of target particles (target particle concentrated liquid) and a liquid from which the target particles have been almost completely removed, and discharges a significantly reduced amount of target particle concentrated liquid compared to the sample liquid.The second separation section then processes this target particle concentrated liquid, thereby significantly reducing the processing time used by the second separation section and enabling large amounts of sample liquid to be processed quickly and in a short period of time.

[0015] Furthermore, in the particle separation device according to the present invention, The second separation section is a target particle concentrated liquid inlet into which the target particle concentrated liquid discharged from the target particle concentrated liquid outlet of the first separation unit is introduced; a buffer solution inlet through which a buffer solution is introduced; a DLD flow path section including a DLD microchannel structure, which allows the target particle concentrated solution introduced from the target particle concentrated solution inlet and the buffer solution introduced from the buffer solution inlet to flow in parallel so as to come into contact with each other, thereby separating the target particle concentrated solution into a buffer solution containing the target particles and a solution from which the target particles have been almost completely removed; an outlet for discharging a liquid from which the target particles separated in the DLD flow path portion have been substantially removed; an outlet for discharging a buffer solution containing the target particles separated in the DLD flow path; The device may include:

[0016] According to the above configuration, the second separation section can transfer the target particles contained in the target particle concentrated solution, which is significantly reduced in volume compared to the sample liquid, to the buffer liquid, thereby separating the target particles into a buffer liquid containing the target particles and a liquid from which the target particles have been almost completely removed.

[0017] Furthermore, the particle separation device according to the present invention may include a sample liquid storage section that supplies the sample liquid to the first separation section, a target particle concentrated liquid storage section that is interposed between the first separation section and the second separation section, stores the target particle concentrated liquid discharged from the target particle concentrated liquid outlet of the first separation section and supplies the target particle concentrated liquid to the second separation section, a target particle removal liquid recovery section that stores the liquid discharged from the target particle removal liquid outlet of the first separation section, and a buffer liquid storage section that supplies the buffer liquid to the second separation section, and may maintain a state in which the air pressure in the sample liquid storage section is higher than the air pressure in the target particle concentrated liquid storage section, and the air pressure in the target particle concentrated liquid storage section, the air pressure in the target particle removal liquid recovery section, and the air pressure in the buffer liquid storage section are approximately equal.

[0018] According to the above configuration, the air pressure difference appropriately controls the introduction of sample liquid into the first separation section, the discharge of target particle concentrated liquid from the first separation section, and the introduction of target particle concentrated liquid into the second separation section, thereby achieving continuous particle separation.

[0019] Furthermore, the particle separation device according to the present invention may include a buffer liquid storage section that supplies the buffer liquid to the second separation section, a liquid level sensor that detects the liquid level of the buffer liquid in the buffer liquid storage section, and a control section that adjusts the amount of the buffer liquid in the buffer liquid storage section by replenishing the buffer liquid from the outside based on the detection result of the liquid level sensor.

[0020] According to the above-mentioned configuration, it becomes possible to maintain an appropriate amount of the buffer liquid in the buffer liquid container.

[0021] Furthermore, in the particle separation device according to the present invention, The second separation section is a target particle concentrated liquid inlet into which the target particle concentrated liquid discharged from the target particle concentrated liquid outlet of the first separation unit is introduced; a DLD flow path section that includes a DLD microchannel structure and that circulates the target particle concentrated solution introduced from the target particle concentrated solution inlet to separate the target particle concentrated solution into a solution with an increased concentration of the target particles and a solution from which the target particles have been substantially removed; an outlet for discharging a liquid having an increased concentration of the target particles separated in the DLD flow path; an outlet for discharging a liquid from which the target particles separated in the DLD flow path portion have been substantially removed; The device may include:

[0022] According to the above configuration, the second separation section can separate the target particle concentrated liquid, the volume of which has been significantly reduced compared to the sample liquid, into a liquid with an even higher concentration of target particles and a liquid from which the target particles have been almost completely removed.

[0023] Furthermore, the particle separation device of the present invention may include a sample liquid storage section that supplies the sample liquid to the first separation section, a target particle concentrated liquid storage section that is interposed between the first separation section and the second separation section, stores the target particle concentrated liquid discharged from the target particle concentrated liquid outlet of the first separation section and supplies the target particle concentrated liquid to the second separation section, and a target particle removal liquid recovery section that stores the liquid discharged from the target particle removal liquid outlet of the first separation section, and may maintain a state in which the air pressure in the sample liquid storage section is higher than the air pressure in the target particle concentrated liquid storage section and the air pressure in the target particle removal liquid recovery section are approximately equal.

[0024] According to the above configuration, the air pressure difference appropriately controls the introduction of sample liquid into the first separation section, the discharge of target particle concentrated liquid from the first separation section, and the introduction of target particle concentrated liquid into the second separation section, thereby achieving continuous particle separation.

[0025] Furthermore, the particle separation device of the present invention may include a target particle concentrated liquid storage section interposed between the first separation section and the second separation section, storing the target particle concentrated liquid discharged from the target particle concentrated liquid outlet of the first separation section and supplying the target particle concentrated liquid to the second separation section, a liquid level sensor that detects the liquid level of the target particle concentrated liquid in the target particle concentrated liquid storage section, and a control section that adjusts the amount of the target particle concentrated liquid in the target particle concentrated liquid storage section based on the detection result of the liquid level sensor.

[0026] According to the above configuration, it becomes possible to maintain an appropriate amount of the target particle concentrated liquid in the target particle concentrated liquid storage section.

[0027] Furthermore, the particle separation device of the present invention may include a sample liquid storage section that supplies the sample liquid to the first separation section, a liquid level sensor that detects the liquid level of the sample liquid in the sample liquid storage section, and a control section that adjusts the amount of the sample liquid in the sample liquid storage section by refilling the sample liquid from the outside based on the detection result of the liquid level sensor.

[0028] According to the above configuration, it becomes possible to maintain an appropriate amount of sample liquid in the sample liquid storage section.

[0029] Furthermore, in the particle separation device according to the present invention, the sample liquid may contain non-target particles having a size different from that of the target particles.

[0030] According to the above configuration, even if the sample liquid contains non-target particles having a size different from that of the target particles, the target particles can be separated.

[0031] Furthermore, in the particle separation device according to the present invention, the target particles may be cells, and the sample liquid may contain circulating tumor cells as the target particles.

[0032] According to the above configuration, cells such as circulating tumor cells (CTCs) can be suitably separated. [Brief description of the drawings]

[0033] [Figure 1] 1 is a diagram showing an example of the configuration of a particle separator (CTC separator) according to a first embodiment of the present invention. [Diagram 2] 1 is a schematic diagram showing the principle of particle separation in a DLD microchannel according to the present invention. FIG. [Diagram 3] 1 is a diagram showing an example of the configuration of a first separation section used in a particle separator (CTC separator) in a first embodiment of the present invention. [Figure 4] 2 is a diagram showing an example of the configuration of a DLD flow path section in a first separation section used in a particle separation device (CTC separation device) in a first embodiment of the present invention. FIG. [Diagram 5] 2 is a diagram showing an example of the configuration of a second separation section used in the particle separation device (CTC separation device) in the first embodiment of the present invention. FIG. [Figure 6] 1 is a diagram showing a variation of the configuration of a particle separator (CTC separator) according to a first embodiment of the present invention. FIG. [Figure 7] FIG. 11 is a diagram showing an example of the configuration of a particle separator (CTC separator) according to a second embodiment of the present invention. [Figure 8] FIG. 11 is a diagram showing an example of the configuration of a first separation section used in a particle separator (CTC separator) according to a second embodiment of the present invention. [Figure 9] FIG. 11 is a diagram showing a variation of the configuration of a particle separator (CTC separator) according to a second embodiment of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0034] Hereinafter, an embodiment of the present invention will be described with reference to the drawings.

[0035] Below, we will explain an embodiment of a particle separation device to which the present invention is applied, for separating (fractionating) target particles from a sample liquid in which target particles and non-target particles of a different size from the target particles are dispersed, and a CTC separation device that uses a blood-derived liquid containing blood cells as non-target particles and circulating tumor cells (CTCs) as target particles as a sample liquid and separates circulating tumor cells from the sample liquid.

[0036] However, the present invention is not limited to separating circulating tumor cells in the blood, but can be applied to separating cells related to target particles from body fluids (including blood, lymph, saliva, urine, tears, etc.) that contain cells other than circulating tumor cells as target particles and cells of different sizes from the target particles as non-target particles. In addition, the target particles and non-target particles are not limited to cells, and the present invention can be widely applied to separating particles related to target particles from a liquid in which two or more types of particles of different sizes are dispersed. Note that the term "dispersion" as used herein includes not only the case where a particle (cell) is suspended in a liquid as a single particle, but also the case where a part or all of the particle is suspended as a cluster, and also includes not only the case where the particle is suspended in a liquid by scattering, but also the case where the particle is settled to some extent. The particle size of the target particles and non-target particles is about 0.1 to 1000 μm.

[0037] Furthermore, in the present invention, the sample liquid may contain target particles to be separated, and does not necessarily contain non-target particles having a size different from that of the target particles. For example, the present invention can be applied to the exchange of a buffer liquid containing particles of a single size or multiple types of particles having a particle size of a certain size or more (a buffer liquid containing particles is used as a sample liquid, and the particles are transferred to another buffer liquid having a different or the same components as the sample liquid) or to the concentration of particles. The term "concentration" as used here is one aspect of particle separation, and means separating a liquid such as a sample liquid containing target particles into a liquid with an increased concentration of target particles and a liquid from which the target particles have been almost removed, and recovering the liquid with an increased concentration of target particles. In addition, "target particles" means particles to be separated, and includes not only particles to be separated and recovered for some use (e.g., testing), but also particles to be separated and removed (removed) because they are unnecessary. In addition, "a liquid from which the target particles have been almost completely removed" means a liquid that is substantially free of target particles (a liquid that contains only a small amount of target particles, or ideally, a liquid that contains no target particles at all), and can be expressed as "a liquid in which the concentration of target particles has decreased" in comparison with "a liquid in which the concentration of target particles has increased."

[0038] First Embodiment A basic concept according to a first embodiment of the present invention will be described with reference to Fig. 1. Fig. 1 is a diagram showing an example of the configuration of a particle separator (CTC separator) according to the first embodiment of the present invention.

[0039] As shown in Figure 1, the particle separation device 10 in this embodiment is roughly configured to include a sample liquid storage section 11, a first separation section 20, a target particle concentrated liquid storage section 12, a target particle removal liquid recovery section 13, a buffer liquid storage section 14, a second separation section 30, a second target particle removal liquid recovery section 15, and a buffer liquid recovery section 16.

[0040] The sample liquid storage section 11 is a container that stores the sample liquid Sa containing the target particles to be separated, and can be formed, for example, by a syringe barrel, etc. The sample liquid Sa may be any liquid that contains the target particles to be separated, but when the target particles are circulating tumor cells in the blood, for example, blood containing circulating tumor cells to which EDTA (ethylenediaminetetraacetic acid) has been added at a final concentration of 2 mM as an anticoagulant can be used.

[0041] The storage space of the sample liquid storage unit 11 can be made airtight. Furthermore, as shown in Fig. 1, the air pressure (air pressure A) in the sample liquid storage unit 11 can be controlled by a pressure generator (air pressure pump) (not shown) or the like that is in communication with the storage space of the sample liquid storage unit 11. As an example, a lid (adapter that seals the upper part of the barrel) through which an air supply tube connected to a pressure generator (not shown) penetrates can be removably attached to an opening located above the sample liquid storage unit 11, and the lid can be attached to the sample liquid storage unit 11 that stores the sample liquid Sa, thereby making it possible to make the storage space of the sample liquid storage unit 11 airtight and to control the air pressure A in the sample liquid storage unit 11.

[0042] Further, the sample liquid storage section 11 is provided with an outlet for discharging the sample liquid Sa at a position where the stored sample liquid Sa can be discharged (for example, at the bottom surface of the sample liquid storage section 11). The outlet of the sample liquid storage section 11 is connected to one end of the first pipeline 41.

[0043] The first separation section 20 is designed based on the basic structure of the DLD method, and has a DLD microchannel structure in which a plurality of fine pillars (micropillars) are arranged (see FIG. 2 described later).

[0044] The first separation section 20 has a sample liquid inlet 21 as a liquid inlet, and has a target particle concentrated liquid outlet 22 and a target particle removal liquid outlet 23 as liquid outlets. The sample liquid inlet 21 is connected to the other end of the first pipe 41, so that the sample liquid Sa discharged from the outlet of the sample liquid storage section 11 is introduced into the sample liquid inlet 21 through the first pipe 41. In addition, the target particle concentrated liquid outlet 22 is connected to one end of the second pipe 42, and the target particle removal liquid outlet 23 is connected to one end of the third pipe 43.

[0045] The first separation section 20 circulates the sample liquid Sa introduced from the sample liquid inlet 21, and separates it into a liquid with an increased concentration of target particles and a liquid from which the target particles have been almost completely removed. The liquid with the increased concentration of target particles is discharged from a target particle concentrated liquid outlet 22 as a target particle concentrated liquid Sb, and the liquid from which the target particles have been almost completely removed is discharged from a target particle removed liquid outlet 23 as a target particle removed liquid Sc. The first separation section 20 functions as a concentration chip that concentrates target particles, and can discharge a target particle concentrated liquid Sb that is significantly reduced in amount compared to the sample liquid Sa. The detailed configuration and function of the first separation section 20 will be described later with reference to Figs. 3 and 4.

[0046] The target particle concentrated liquid storage unit 12 is a container that stores the target particle concentrated liquid Sb discharged from the first separation unit 20, and can be formed by, for example, a barrel of a syringe. The other end of the second pipeline 42 is connected to the storage space of the target particle concentrated liquid storage unit 12, so that the target particle concentrated liquid Sb discharged from the target particle concentrated liquid outlet 22 of the first separation unit 20 is introduced into the target particle concentrated liquid storage unit 12 through the second pipeline 42.

[0047] The storage space of the target particle concentrate storage unit 12 can be made airtight. Furthermore, as shown in Fig. 1, the air pressure (air pressure B) in the target particle concentrate storage unit 12 can be controlled by a pressure generator (air pressure pump) (not shown) or the like that is connected to the storage space of the target particle concentrate storage unit 12. As an example, a lid (adapter that seals the upper part of the barrel) through which an air supply tube connected to a pressure generator (not shown) and a second pipe 42 penetrate can be removably attached to an opening located above the target particle concentrate storage unit 12, and the lid can be attached to the target particle concentrate storage unit 12 to make the storage space of the target particle concentrate storage unit 12 airtight and control the air pressure B of the target particle concentrate storage unit 12.

[0048] Further, the target particle concentrated liquid storage section 12 is provided with an outlet for discharging the target particle concentrated liquid Sb at a position where the accommodated target particle concentrated liquid Sb can be discharged (for example, the bottom surface of the target particle concentrated liquid storage section 12). The outlet of the target particle concentrated liquid storage section 12 is connected to one end of the fourth pipe 44.

[0049] The target particle removal liquid recovery section 13 is a container that contains the target particle removal liquid Sc discharged from the first separation section 20, and can be formed by, for example, a barrel of a syringe. The other end of the third pipeline 43 is connected to the storage space of the target particle removal liquid recovery section 13, so that the target particle removal liquid Sc discharged from the target particle removal liquid outlet 23 of the first separation section 20 is introduced into the target particle removal liquid recovery section 13 through the third pipeline 43.

[0050] The storage space of the target particle removal liquid recovery section 13 can be made airtight. Furthermore, as shown in Fig. 1, the air pressure (air pressure B) in the target particle removal liquid recovery section 13 can be controlled by a pressure generator (air pressure pump) (not shown) or the like that is in communication with the storage space of the target particle removal liquid recovery section 13. As an example, a lid (adapter that seals the upper part of the barrel) through which an air supply tube connected to a pressure generator (not shown) and a third pipe line 43 penetrate can be removably attached to an opening located above the target particle removal liquid recovery section 13, and by attaching this lid to the target particle removal liquid recovery section 13, the storage space of the target particle removal liquid recovery section 13 can be made airtight and the air pressure B in the target particle removal liquid recovery section 13 can be controlled.

[0051] The buffer liquid storage section 14 is a container that stores the buffer liquid Ba, and can be configured, for example, by the barrel of a syringe. The buffer liquid Ba is not particularly limited as long as it is a liquid, and for example, PBS or glycerin-containing PBS can be used. Note that, since a liquid with a buffering effect is often used, it is called a buffer liquid for convenience, but the "buffer liquid" in this specification also includes liquids without a buffering effect.

[0052] The storage space of the buffer liquid storage unit 14 can be made airtight. Furthermore, as shown in Fig. 1, the air pressure (air pressure B) in the buffer liquid storage unit 14 can be controlled by a pressure generator (air pressure pump) (not shown) or the like that is in communication with the storage space of the buffer liquid storage unit 14. As an example, a lid (adapter that seals the upper part of the barrel) through which an air supply tube connected to a pressure generator (not shown) penetrates can be removably attached to an opening located above the buffer liquid storage unit 14, and the lid can be attached to the buffer liquid storage unit 14 in which the buffer liquid Ba is stored, thereby making it possible to make the storage space of the buffer liquid storage unit 14 airtight and to control the air pressure B in the buffer liquid storage unit 14.

[0053] The buffer liquid storage unit 14 is provided with an outlet for discharging the buffer liquid Ba at a position where the stored buffer liquid Ba can be discharged (for example, at the bottom surface of the buffer liquid storage unit 14). The outlet of the buffer liquid storage unit 14 is connected to one end of a fifth pipe line 45.

[0054] The second separation section 30 is designed based on the basic structure of the DLD method, and has a DLD microchannel structure in which a plurality of fine pillars (micropillars) are arranged (see FIG. 2 described later).

[0055] The second separation section 30 includes a target particle concentrated liquid inlet 31 and a buffer liquid inlet 32 ​​as liquid inlets, and a second target particle removal liquid outlet 33 and a buffer liquid outlet 34 as liquid outlets. The target particle concentrated liquid inlet 31 is connected to the other end of a fourth pipe 44, so that the target particle concentrated liquid Sb discharged from the outlet of the target particle concentrated liquid storage section 12 is introduced into the target particle concentrated liquid inlet 31 through the fourth pipe 44. The buffer liquid inlet 32 ​​is connected to the other end of a fifth pipe 45, so that the buffer liquid Ba discharged from the outlet of the buffer liquid storage section 14 is introduced into the buffer liquid inlet 32 ​​through the fifth pipe 45. The second target particle removal liquid outlet 33 is connected to one end of a sixth pipe 46, and the buffer liquid outlet 34 is connected to one end of a seventh pipe 47.

[0056] The second separation section 30 separates the target particle concentrated solution Sb introduced from the target particle concentrated solution inlet 31 and the buffer solution Ba introduced from the buffer solution inlet 32 ​​into a buffer solution containing the target particles and a solution from which the target particles have been almost completely removed by circulating the target particle concentrated solution Sb introduced from the target particle concentrated solution inlet 31 in parallel with the buffer solution Ba introduced from the buffer solution inlet 32. The solution from which the target particles have been almost completely removed is discharged from the second target particle removal solution outlet 33 as a second target particle removal solution Sd, and the buffer solution containing the target particles is discharged from the buffer solution outlet 34 as a recovery buffer solution Bb. The second separation section 30 functions as a sorting chip that extracts the target particles into the recovery buffer solution Bb. The detailed configuration and function of the second separation section 30 will be described later with reference to FIG. 5.

[0057] The second target particle removal liquid recovery section 15 is a container that contains the second target particle removal liquid Sd discharged from the second separation section 30. The other end of the sixth pipeline 46 is connected to the storage space of the second target particle removal liquid recovery section 15, and the second target particle removal liquid Sd discharged from the second target particle removal liquid outlet 33 of the second separation section 30 is introduced into the second target particle removal liquid recovery section 15 through the sixth pipeline 46. The storage space of the second target particle removal liquid recovery section 15 does not need to be airtight, and the air pressure inside the second target particle removal liquid recovery section 15 may be equal to atmospheric pressure.

[0058] The buffer liquid recovery section 16 is a container that contains the recovered buffer liquid Bb discharged from the second separation section 30. The other end of the seventh pipe 47 is connected to the storage space of the buffer liquid recovery section 16, and the recovered buffer liquid Bb discharged from the buffer liquid outlet 34 of the second separation section 30 is introduced into the buffer liquid recovery section 16 through the seventh pipe 47. The recovered buffer liquid Bb contained in the buffer liquid recovery section 16 can be used, for example, to measure the number of target particles in the sample liquid Sa. The storage space of the buffer liquid recovery section 16 does not need to be airtight, and the air pressure inside the buffer liquid recovery section 16 may be equal to atmospheric pressure.

[0059] A description will now be given of the configurations and functions of the first separation section 20 and the second separation section 30. Fig. 2 is a schematic diagram showing the principle of particle separation in the DLD microchannel according to the present invention.

[0060] The first separation section 20 and the second separation section 30 are designed based on the basic structure of the DLD method. The first separation section 20 and the second separation section 30 each have a DLD microchannel structure in which a plurality of fine pillars (micropillars) are arranged based on the principle of the DLD method as described in Non-Patent Document 1, for example.

[0061] According to the DLD method, when a particle dispersion is flowed through a group of pillars P arranged according to a predetermined rule as shown in Fig. 2, small particles proceed along the direction of the flow, while large particles proceed obliquely to the direction of the flow, as they are prevented from proceeding along the direction of the flow by the presence of the pillars P. By appropriately setting a threshold value determined by the interval G of the pillars P and the shift amount d, it is possible to separate particles with diameters less than the threshold value from particles with diameters equal to or greater than the threshold value.

[0062] The configuration and function of the first separation section 20 will be described with reference to Figures 3 and 4. Figure 3 is a diagram showing an example of the configuration of the first separation section used in the particle separation device (CTC separation device) in the first embodiment of the present invention. Figure 4 is a diagram showing an example of the configuration of the DLD flow path section in the first separation section used in the particle separation device (CTC separation device) in the first embodiment of the present invention.

[0063] As shown in Figure 3, the first separation section 20 is configured with one sample liquid inlet 21, two outlets (a target particle concentrated liquid outlet 22 and a target particle removed liquid outlet 23), a DLD flow path section 24, and a confluence flow path 25.

[0064] The DLD flow path section 24 has a DLD micro-flow path structure in which a plurality of fine pillars are arranged. A sample liquid inlet 21 is arranged on the upstream side of the DLD flow path section 24, and the sample liquid Sa containing the target particles introduced from the sample liquid inlet 21 can flow through the DLD micro-flow path formed in the DLD flow path section 24.

[0065] As shown in FIG. 4, multiple (e.g., 10) DLD microchannels (including mirror image DLD microchannels) are arranged in parallel within the DLD flow channel section 24. In each DLD microchannel, target particles with a relatively large diameter (e.g., circulating tumor cells of about 12 to 15 μm) contained in the sample liquid Sa proceed obliquely with respect to the flow direction of the sample liquid Sa, and move so as to gather on one side (the opposite side in the mirror image) perpendicular to the flow direction of the sample liquid Sa. As a result, the target particles gather at a predetermined position on the downstream side of each DLD microchannel, and the sample liquid Sa is separated into a liquid with an increased concentration of target particles and a liquid from which the target particles have been almost completely removed.

[0066] The confluence channel 25 is a channel for confluence of liquids with increased concentrations of target particles. The liquids with increased concentrations of target particles obtained by the flow of the sample liquid in each DLD microchannel are confluenced through the confluence channel 25 and discharged as a target particle concentrated liquid Sb from one of the two discharge ports (target particle concentrated liquid discharge port 22).

[0067] On the other hand, the liquid from which the target particles have been almost completely removed, obtained by the flow of sample liquid in each DLD microchannel, joins together in the DLD channel section 24 and is discharged as target particle removal liquid Sc from the other of the two outlets (target particle removal liquid outlet 23).

[0068] In this way, the first separation unit 20 functions as a concentration chip that concentrates the target particles, and can discharge a target particle concentrated solution Sb that is significantly reduced in amount compared to the sample liquid Sa. For example, the first separation unit 20 can reduce the amount of the target particle concentrated solution Sb to about 1 / 10 of the amount of the sample liquid Sa introduced from the sample liquid inlet 21.

[0069] The first separation unit 20 is configured to introduce only the sample liquid Sa, and the entire DLD microchannel can be used for the distribution (processing) of the sample liquid Sa, so the processing speed of the sample liquid Sa can be increased compared to a configuration in which two types of liquid (sample liquid Sa and buffer liquid) are introduced. In addition, the first separation unit 20 is configured to introduce only the sample liquid Sa, and only the sample liquid Sa needs to be distributed to each DLD microchannel, so the structure is less likely to become complicated even if the DLD microchannels are parallelized compared to a configuration in which two types of liquid (sample liquid Sa and buffer liquid) are introduced, and further, there is no need to adjust the flow rate of the buffer liquid according to the flow rate of the sample liquid Sa. The first separation unit 20 can process a large amount of sample liquid Sa in a short time and at high speed by parallelizing the DLD microchannels, and the processing speed (flow rate) of the liquid (sample liquid Sa) in the first separation unit 20 is significantly faster than that of the second separation unit 30, and the processing speed is, for example, 2 mL per minute.

[0070] The first separation section 20 incorporated in the particle separation device 10 shown in Figure 1 can discharge target particle concentrated liquid Sb from the target particle concentrated liquid outlet 22 into the target particle concentrated liquid storage section 12 when the sample liquid Sa contained in the sample liquid storage section 11 is introduced into the sample liquid inlet 21, and can discharge the target particle removal liquid Sc from the target particle removal liquid outlet 23 into the target particle removal liquid recovery section 13.

[0071] The configuration and function of the second separation section 30 will be described with reference to Fig. 5. Fig. 5 is a diagram showing an example of the configuration of the second separation section used in the particle separator (CTC separator) in the first embodiment of the present invention.

[0072] As shown in Figure 5, the second separation section 30 is configured to include a target particle concentrated liquid inlet 31, a buffer liquid inlet 32, a second target particle removal liquid outlet 33, a buffer liquid outlet 34, and a DLD flow path section 35.

[0073] The DLD flow path section 35 has a DLD microchannel structure in which a plurality of fine pillars are arranged. A target particle concentrated solution inlet 31 and a buffer solution inlet 32 ​​are arranged on the upstream side of the DLD flow path section 35, so that a target particle concentrated solution Sb containing target particles introduced from the target particle concentrated solution inlet 31 and a buffer solution Ba introduced from the buffer solution inlet 32 ​​can flow through the DLD microchannel formed in the DLD flow path section 35.

[0074] A DLD microchannel is disposed within the DLD flow channel section 35. In the DLD microchannel, target particles having a relatively large diameter (e.g., tumor cells circulating in the blood having a size of about 12 to 15 μm) contained in the target particle concentrated solution Sb proceed obliquely with respect to the flow direction of the target particle concentrated solution Sb, and move to the buffer solution Ba that flows in parallel as a laminar flow while in contact with the target particle concentrated solution Sb. On the other hand, in the DLD microchannel, non-target particles having a relatively small diameter (e.g., blood cells having a size of about 8 μm) contained in the target particle concentrated solution Sb proceed along the flow direction of the target particle concentrated solution Sb. As a result, on the downstream side of the DLD microchannel, the target particle concentrated solution Sb is separated into a recovery buffer solution Bb containing target particles that have moved from the target particle concentrated solution Sb to the buffer solution Ba, and a second target particle removal solution Sd from which the target particles have been almost completely removed.

[0075] The second target particle removal liquid Sd from which the target particles have been almost completely removed and obtained by the liquid flow in the DLD microchannel is discharged from a second target particle removal liquid outlet 33. On the other hand, the recovery buffer liquid Bb containing the target particles and obtained by the liquid flow in the DLD microchannel is discharged from a buffer liquid outlet 34.

[0076] In addition, within the DLD microchannel, the target particle concentrated solution Sb and the buffer solution Ba slightly mix with each other as they flow in parallel as laminar flows while in contact with each other, so that the second target particle removal solution Sd discharged from the second target particle removal solution outlet 33 may contain a portion of the recovery buffer solution Bb, and the recovery buffer solution Bb discharged from the buffer solution outlet 34 may contain a portion of the second target particle removal solution Sd.

[0077] In this way, the second separation section 30 functions as a sorting chip that transfers the target particles contained in the target particle concentrated solution Sb to the buffer solution Ba, thereby extracting the target particles into the recovery buffer solution Bb. The processing speed of the liquid (target particle concentrated solution Sb) in the second separation section 30 is slower than that of the first separation section 20, and the processing speed is, for example, 0.1 mL per minute.

[0078] The second separation section 30 incorporated in the particle separation device 10 shown in Figure 1 can discharge the second target particle removal liquid Sd from the second target particle removal liquid outlet 33 into the second target particle removal liquid recovery section 15 and discharge the recovered buffer liquid Bb from the buffer liquid outlet 34 into the buffer liquid recovery section 16 when the target particle concentrated liquid Sb contained in the target particle concentrated liquid storage section 12 is introduced into the target particle concentrated liquid inlet 31 and the buffer liquid Ba contained in the buffer liquid storage section 14 is introduced into the buffer liquid inlet 32.

[0079] An example of the operation of the particle separator 10 shown in FIG. 1 will be described.

[0080] First, it is preferable to fill all DLD flow path sections and their liquid introduction pipes (first pipe 41, fourth pipe 44, fifth pipe 45) with liquid (e.g., buffer liquid). Thereafter, the sample liquid Sa is accommodated in the sample liquid accommodation section 11, and the buffer liquid Ba is accommodated in the buffer liquid accommodation section 14. When the air pressure A in the sample liquid accommodation section 11 is made higher than the air pressure B (e.g., atmospheric pressure) in the target particle concentrated liquid accommodation section 12, the air pressure B (e.g., atmospheric pressure) in the target particle removal liquid recovery section 13, and the air pressure B (e.g., atmospheric pressure) in the buffer liquid accommodation section 14, the sample liquid Sa accommodated in the sample liquid accommodation section 11 is pressure-fed into the first separation section 20 via the first pipe 41. The sample liquid Sa introduced into the first separation section 20 is separated into a liquid with an increased concentration of target particles (target particle concentrated liquid Sb) and a liquid from which the target particles have been almost completely removed (target particle removed liquid Sc), and then the target particle concentrated liquid Sb is discharged into the target particle concentrated liquid storage section 12, and the target particle removed liquid Sc is discharged into the target particle removed liquid recovery section 13.

[0081] When an appropriate amount of target particle concentrated liquid Sb has accumulated in the target particle concentrated liquid storage section 12, the air pressure B in the target particle concentrated liquid storage section 12, the air pressure B in the target particle removal liquid recovery section 13, and the air pressure B in the buffer liquid storage section 14 are made higher than the air pressure (e.g., atmospheric pressure) in the second target particle removal liquid recovery section 15 and the air pressure (e.g., atmospheric pressure) in the buffer liquid recovery section 16, so that the target particle concentrated liquid Sb stored in the target particle concentrated liquid storage section 12 and the buffer liquid Ba stored in the buffer liquid storage section 14 are pressurized and fed into the second separation section 30 via the fourth pipeline 44 and the fifth pipeline 45, respectively. The target particle concentrated liquid Sb and buffer liquid Ba introduced into the second separation section 30 are separated into a buffer liquid containing target particles (recovery buffer liquid Bb) and a liquid from which the target particles have been almost completely removed (second target particle removal liquid Sd), and then the second target particle removal liquid Sd is discharged into the second target particle removal liquid recovery section 15, and the recovery buffer liquid Bb is discharged into the buffer liquid recovery section 16.

[0082] The pressure in the second target particle removal liquid recovery section 15 and the pressure in the buffer liquid recovery section 16 are set to, for example, atmospheric pressure, and the pressure B in the target particle concentrated liquid storage section 12, the pressure B in the target particle removal liquid recovery section 13 and the pressure B in the buffer liquid storage section 14 are set higher than atmospheric pressure (pressure B>atmospheric pressure), and further, the pressure A in the sample liquid storage section 11 is set higher than pressure B (pressure A>pressure B), so that the liquid in each storage section is continuously pumped in a direction in which it will be subjected to the particle separation action, thereby realizing continuous particle separation. As an example, the pressures A and B are set to 1.5 to 2.5 atmospheres (where pressure A>pressure B).

[0083] By making the air pressure inside the target particle concentrated liquid storage unit 12 and the air pressure inside the target particle removal liquid recovery unit 13 equal (air pressure B), the target particle concentrated liquid Sb and the target particle removal liquid Sc are discharged from the first separation unit 20 without bias. Furthermore, by making the air pressure inside the target particle concentrated liquid storage unit 12 and the air pressure inside the buffer liquid storage unit 14 equal (air pressure B), the buffer liquid Ba and the target particle concentrated liquid Sb can be introduced into the second separation unit 30 at the same flow rate. The air pressure inside the target particle concentrated liquid storage unit 12, the air pressure inside the target particle removal liquid recovery unit 13, and the air pressure inside the buffer liquid storage unit 14 may be configured to be equal (air pressure B) by connecting the target particle concentrated liquid storage unit 12, the target particle removal liquid recovery unit 13, and the buffer liquid storage unit 14 to the same pressure generator. Also, the target particle concentrated liquid storage unit 12, the target particle removal liquid recovery unit 13, and the buffer liquid storage unit 14 may be connected by a pipeline through which gas flows to maintain equal pressure, and a pressure generator may be connected to the pipeline to control the atmospheric pressure B. Furthermore, the target particle concentrated liquid storage unit 12, the target particle removal liquid recovery unit 13, and the buffer liquid storage unit 14 may be connected by a pipeline through which gas flows to maintain equal pressure, and the target particle concentrated liquid Sb may be discharged from the first separation unit 20 to the target particle concentrated liquid storage unit 12, and the target particle removal liquid Sc may be discharged to the target particle removal liquid recovery unit 13, respectively, to indirectly increase the atmospheric pressure B.

[0084] 1, the pressure A in the sample liquid storage section 11, the pressure B in the target particle concentrated liquid storage section 12, the pressure B in the target particle removal liquid recovery section 13, and the pressure B in the buffer liquid storage section 14 can all be set to atmospheric pressure (pressure A = pressure B = atmospheric pressure). When pressure A and pressure B are set to atmospheric pressure, the pressure difference between the upstream and downstream sides disappears, and the pumping of the liquid can be stopped.

[0085] The particle separation device 10 shown in FIG. 1 is configured to perform two-stage processing, processing in a first separation section 20 and processing in a second separation section 30. The first separation section 20 can obtain a target particle concentrated liquid Sb from the sample liquid Sa at a high processing speed. The target particle concentrated liquid Sb is a liquid in which the amount of the target particle concentrated liquid Sb is significantly reduced compared to the sample liquid Sa. After obtaining the target particle concentrated liquid Sb in a significantly reduced amount compared to the sample liquid Sa in the first separation section 20, the obtained target particle concentrated liquid Sb is introduced into the second separation section 30, whereby the time required for processing in the second separation section 30 (sorting of target particles) is significantly shortened, and a large amount of sample liquid can be processed quickly and at high speed.

[0086] Furthermore, in this embodiment, a liquid level sensor 51 may be optionally provided that detects the liquid level of the target particle concentrated liquid Sb contained in the target particle concentrated liquid storage section 12. The liquid level sensor 51 is not particularly limited as long as it can detect the liquid level of the target particle concentrated liquid Sb, and for example, a non-contact sensor that can irradiate light, ultrasonic waves, or the like from outside the target particle concentrated liquid storage section 12 and measure the liquid level of the target particle concentrated liquid Sb from the transmitted light, reflected waves, or the like may be used, and multiple sensors may be provided.

[0087] By monitoring the liquid level of the target particle concentrate Sb detected by the liquid level sensor 51, the liquid level of the target particle concentrate Sb contained in the target particle concentrate storage section 12 can be maintained at a constant level, and the target particle concentrate Sb can be maintained at an appropriate amount. For example, when the liquid level sensor 51 detects that the liquid level of the target particle concentrate Sb has fallen below a predetermined level (a drop threshold), the amount of the target particle concentrate Sb may be increased. In this case, the liquid level of the target particle concentrate Sb can be increased by increasing the air pressure A in the sample liquid storage section 11, decreasing the air pressure B in the target particle concentrate storage section 12, or both, thereby increasing the difference between the air pressure A and the air pressure B. Also, for example, when the liquid level sensor 51 detects that the liquid level of the target particle concentrate Sb has risen above a predetermined level (a rise threshold), the amount of the target particle concentrate Sb may be decreased. In this case, the liquid level of the target particle concentrated liquid Sb can be lowered by lowering the air pressure A in the sample liquid storage section 11, increasing the air pressure B in the target particle concentrated liquid storage section 12, or both, thereby reducing the difference between air pressure A and air pressure B.

[0088] The method of adjusting the air pressure A and / or the air pressure B to raise or lower the liquid level of the target particle concentrated liquid Sb is not particularly limited. As an example, the air pressure supplied from the pressure generator may be adjusted manually, or the air pressure supplied from the pressure generator may be automatically adjusted according to the liquid level detected by the liquid level sensor 51. In addition, preferably, when adjusting the air pressure B in the target particle concentrated liquid storage section 12, the air pressure B in the target particle removal liquid recovery section 13 and the air pressure B in the buffer liquid storage section 14 may be adjusted simultaneously and in the same manner so that the air pressures in the target particle concentrated liquid storage section 12, the target particle removal liquid recovery section 13, and the buffer liquid storage section 14 are equal in pressure (air pressure B).

[0089] Next, a variation of this embodiment will be described with reference to Fig. 6. Fig. 6 is a diagram showing a variation of the configuration of the particle separator (CTC separator) in the first embodiment of the present invention. In the description of Fig. 6, the description of the same components as those in Fig. 1 may be simplified or omitted.

[0090] The particle separation device 100 shown in Figure 6 is similar to the particle separation device 10 shown in Figure 1 and is roughly configured to include a sample liquid storage section 11, a first separation section 20, a target particle concentrated liquid storage section 12, a target particle removal liquid recovery section 13, a buffer liquid storage section 14, a second separation section 30, a second target particle removal liquid recovery section 15, and a buffer liquid recovery section 16.

[0091] The particle separator 100 shown in FIG. 6 further includes liquid level sensors 51 to 56, pressure generators 71 and 72, a buffer liquid supply unit 73, a tube pump 74, and a control unit 75.

[0092] 1, and has a function of detecting the liquid level of the target particle concentrated liquid Sb contained in the target particle concentrated liquid storage section 12. The liquid level sensor 51 is connected to the control section 75 via a signal line (not shown), and is configured to be able to output a signal including the detected liquid level of the target particle concentrated liquid Sb to the control section 75. As described above, the liquid level sensor 51 is not particularly limited as long as it can detect the liquid level of the target particle concentrated liquid Sb, and for example, a non-contact sensor can be used, and multiple sensors may be provided.

[0093] The liquid level sensor 52 has a function of detecting the liquid level of the buffer liquid Ba contained in the buffer liquid container 14. The liquid level sensor 52 is connected to the control unit 75 via a signal line (not shown), and is configured to be able to output a signal including the detected liquid level of the buffer liquid Ba to the control unit 75. The liquid level sensor 52 is not particularly limited as long as it can detect the liquid level of the buffer liquid Ba, and for example, a non-contact sensor can be used, and multiple sensors may be provided.

[0094] The liquid level sensor 53 has a function of detecting the liquid level of the sample liquid Sa contained in the sample liquid storage section 11. The liquid level sensor 53 is connected to the control section 75 via a signal line (not shown), and is configured to be able to output a signal including the detected liquid level of the sample liquid Sa to the control section 75. The liquid level sensor 53 is not particularly limited as long as it can detect the liquid level of the sample liquid Sa, and for example, a non-contact type sensor can be used, and multiple sensors may be provided.

[0095] The liquid level sensor 54 has a function of detecting the liquid level of the target particle removal liquid Sc contained in the target particle removal liquid recovery unit 13. The liquid level sensor 54 is connected to the control unit 75 via a signal line (not shown), and is configured to be able to output a signal including the detected liquid level of the target particle removal liquid Sc to the control unit 75. The liquid level sensor 54 is not particularly limited as long as it can detect the liquid level of the target particle removal liquid Sc, and for example, a non-contact sensor can be used, and multiple sensors may be provided.

[0096] The liquid level sensor 55 has a function of detecting the liquid level of the second target particle removal liquid Sd contained in the second target particle removal liquid recovery unit 15. The liquid level sensor 55 is connected to the control unit 75 via a signal line (not shown), and is configured to be able to output a signal including the detected liquid level of the second target particle removal liquid Sd to the control unit 75. The liquid level sensor 55 is not particularly limited as long as it can detect the liquid level of the second target particle removal liquid Sd, and for example, a non-contact sensor can be used, and multiple sensors may be provided.

[0097] The liquid level sensor 56 has a function of detecting the liquid level of the recovered buffer liquid Bb contained in the buffer liquid recovery unit 16. The liquid level sensor 56 is connected to the control unit 75 via a signal line (not shown), and is configured to be able to output a signal including the detected liquid level of the recovered buffer liquid Bb to the control unit 75. The liquid level sensor 56 is not particularly limited as long as it can detect the liquid level of the recovered buffer liquid Bb, and for example, a non-contact type sensor can be used, and multiple sensors may be provided.

[0098] The pressure generator 71 is a device capable of controlling the air pressure (air pressure A) in the sample liquid storage section 11 to a desired air pressure. The pressure generator 72 is a device capable of controlling the air pressure B in the target particle concentrated liquid storage section 12, the air pressure B in the target particle removal liquid recovery section 13, and the air pressure B in the buffer liquid storage section 14 to be equal and to the desired air pressures. The pressure generators 71 and 72 are connected to a control section 75 via signal lines 81 and 82, respectively, and are configured to be operable and controlled by the control section 75. The pressure generators 71 and 72 are not particularly limited as long as they can generate compressed air, and may be an electric rotary pump having a rotor as a moving part, an electric diaphragm pump having a diaphragm as a moving part, an electric plunger pump having a plunger as a moving part, an electric piston pump having a piston as a moving part, or the like.

[0099] The buffer solution supplying unit 73 is a device capable of refilling the buffer solution storage unit 14 with the buffer solution Ba. The buffer solution supplying unit 73 is connected to the control unit 75 via a signal line 83, and is configured so that its operation can be controlled by the control unit 75. There are no particular limitations on the buffer solution supplying unit 73 as long as it can refill the buffer solution storage unit 14 with the buffer solution Ba.

[0100] The tube pump 74 is a device capable of replenishing the sample liquid Sa in the sample liquid storage section 11. The tube pump 74 is connected to the control section 75 via a signal line 84, and is configured so that its operation can be controlled by the control section 75. The tube pump 74 is configured, for example, with a tube through which the sample liquid Sa can pass, and a rotor equipped with a roller that rotates while squashing the tube, and is configured so that the sample liquid Sa in the tube can be sent into the sample liquid storage section 11 by the continuous rotation of the rotor.

[0101] The control unit 75 is connected to the liquid level sensors 51 to 56, the pressure generators 71 and 72, the buffer liquid supply unit 73, and the tube pump 74, and is configured to adjust the amount of liquid in each of the storage units or recovery units by controlling the operation of the pressure generators 71 and 72, the buffer liquid supply unit 73, and the tube pump 74 based on the liquid level of the liquid in each of the storage units or recovery units supplied from the liquid level sensors 51 to 56. The control unit 75 is configured by a processor such as a CPU, and can control the pressure generators 71 and 72, the buffer liquid supply unit 73, and the tube pump 74 to perform desired operations by executing program instructions that specify the operation control procedures of the pressure generators 71 and 72, the buffer liquid supply unit 73, and the tube pump 74 according to the liquid level of the liquid in each of the storage units or recovery units. The particle separation device 100 shown in FIG. 6 may be provided with a plurality of control units 75 corresponding to each of the liquid level sensors 51 to 56 and each device to be controlled.

[0102] An example of the operation of the particle separator 100 shown in FIG. 6 will be described.

[0103] In the particle separation device 100 shown in Fig. 6, the process of obtaining a recovery buffer solution Bb containing target particles by performing particle separation from a sample solution Sa is similar to that of the particle separation device 10 shown in Fig. 1. That is, in the particle separation device 100 shown in Fig. 6, similar to the particle separation device 10 shown in Fig. 1, when the sample solution Sa contained in the sample solution container 11 is introduced into the first separation section 20, a target particle concentrated solution Sb and a target particle removal solution Sc are discharged, and further, when the target particle concentrated solution Sb and the buffer solution Ba are introduced into the second separation section 30, a second target particle removal solution Sd and a recovery buffer solution Bb can be discharged.

[0104] The liquid level sensor 51 detects the liquid level of the concentrated liquid Sb of target particles at a predetermined period, and outputs a signal including the detected liquid level to the control unit 75.

[0105] When the liquid level sensor 51 detects that the liquid level of the target particle concentrated liquid Sb has dropped below a predetermined level (drop threshold), the control unit 75 instructs the pressure generator 71 to increase the atmospheric pressure A to a predetermined value, instructs the pressure generator 72 to decrease the atmospheric pressure B to a predetermined value, or performs both, thereby increasing the difference between the atmospheric pressure A and the atmospheric pressure B. As a result, the liquid level of the target particle concentrated liquid Sb can be increased.

[0106] Furthermore, when the liquid level sensor 51 detects that the liquid level of the target particle concentrated liquid Sb has risen above a predetermined level (rising threshold), the control unit 75 instructs the pressure generator 71 to lower the atmospheric pressure A to a predetermined value, instructs the pressure generator 72 to raise the atmospheric pressure B to a predetermined value, or performs both, thereby reducing the difference between the atmospheric pressure A and the atmospheric pressure B. As a result, the liquid level of the target particle concentrated liquid Sb can be lowered.

[0107] In this way, the control unit 75 controls the operation of the pressure generating devices 71, 72 based on the liquid level detection results by the liquid level sensor 51, eliminating the need for the operator to monitor constantly and making it possible to maintain an appropriate amount of target particle concentrated liquid Sb in the target particle concentrated liquid storage section 12.

[0108] The liquid level sensor 52 detects the liquid level of the buffer liquid Ba at a predetermined period, and outputs a signal including the detected liquid level to the control unit 75.

[0109] When the liquid level sensor 52 detects that the liquid level of the buffer liquid Ba has dropped below a predetermined level (lower threshold), the control unit 75 instructs the buffer liquid supply unit 73 to replenish a certain amount of buffer liquid Ba. As a result, the liquid level of the buffer liquid Ba can be raised. The control unit 75 may control the operation of the pressure generators 71 and 72 to set the atmospheric pressure A and the atmospheric pressure B to atmospheric pressure and temporarily suspend the operation of the particle separator 100, and then instruct the buffer liquid supply unit 73 to replenish the buffer liquid Ba. Alternatively, the control unit 75 may alert the operator by outputting a warning sound or emitting light, etc., to have the operator replenish the buffer liquid Ba.

[0110] In this way, the control unit 75 controls the operation of the buffer liquid supply unit 73 based on the liquid level detection results by the liquid level sensor 52, eliminating the need for the operator to monitor the operation constantly and making it possible to maintain an appropriate amount of buffer liquid Ba in the buffer liquid storage unit 14.

[0111] The liquid level sensor 53 detects the liquid level of the sample liquid Sa at a predetermined period, and outputs a signal including the detected liquid level to the control unit 75.

[0112] When the liquid level sensor 53 detects that the liquid level of the sample liquid Sa has dropped below a predetermined level (lower threshold), the control unit 75 instructs the tube pump 74 to replenish a certain amount of sample liquid Sa. As a result, the liquid level of the sample liquid Sa can be raised. Note that the control unit 75 may control the operation of the pressure generators 71, 72 to set the atmospheric pressure A and the atmospheric pressure B to atmospheric pressure and temporarily suspend the operation of the particle separator 100, and then instruct the tube pump 74 to replenish the sample liquid Sa, or may alert the operator by outputting a warning sound or emitting light, etc., to have the operator replenish the sample liquid Sa.

[0113] In this way, the control unit 75 controls the operation of the tube pump 74 based on the liquid level detection results by the liquid level sensor 53, eliminating the need for the operator to monitor constantly and making it possible to maintain an appropriate amount of sample liquid Sa in the sample liquid storage section 11.

[0114] The liquid level sensors 54 to 56 detect the liquid levels of the target particle removal liquid Sc, the second target particle removal liquid Sd, and the collection buffer liquid Bb at predetermined intervals, and output signals including the detected liquid levels to the control unit 75.

[0115] When the liquid level sensor 54 detects that the liquid level of the target particle removal liquid Sc has risen above a predetermined level (upper threshold), the control unit 75 controls a recovery mechanism (not shown) to recover the target particle removal liquid Sc from the target particle removal liquid recovery unit 13. When the liquid level sensor 55 detects that the liquid level of the second target particle removal liquid Sd has risen above a predetermined level (upper threshold), the control unit 75 controls a recovery mechanism (not shown) to recover the second target particle removal liquid Sd from the second target particle removal liquid recovery unit 15. When the liquid level sensor 56 detects that the liquid level of the recovery buffer liquid Bb has risen above a predetermined level (upper threshold), the control unit 75 controls a recovery mechanism (not shown) to recover the recovery buffer liquid Bb from the buffer liquid recovery unit 16. In addition, the control unit 75 may control the operation of the pressure generators 71, 72 to set the air pressure A and air pressure B to atmospheric pressure and temporarily interrupt the operation of the particle separation device 100, and then issue instructions to collect the target particle removal solution Sc, the second target particle removal solution Sd, and the recovery buffer solution Bb, or may alert the operator by emitting a warning sound or emitting light, etc., to cause the operator to collect the target particle removal solution Sc, the second target particle removal solution Sd, and the recovery buffer solution Bb.

[0116] In this manner, by monitoring the liquid levels detected by the liquid level sensors 54 to 56, the target particle removing liquid Sc, the second target particle removing liquid Sd, and the recovery buffer liquid Bb can be recovered at appropriate times.

[0117] 6, the liquid level sensors 51-56 are provided at a total of six locations, but it is not necessary to provide all of the liquid level sensors 51-56, and only one of the liquid level sensors 51-56, or a plurality of liquid level sensors arbitrarily selected from the liquid level sensors 51-56, may be provided. For example, the liquid level sensors 51-53 may be provided, while the liquid level sensors 54-56 may not be provided. In this case, when refilling the buffer liquid Ba or the sample liquid Sa based on the detection results of the liquid level sensors 52, 53, the target particle removal liquid Sc, the second target particle removal liquid Sd, and the recovery buffer liquid Bb may be simultaneously recovered.

[0118] 1, in the particle separation device 100 shown in Fig. 6, a target particle concentrated liquid Sb that is significantly reduced in volume compared to the sample liquid Sa is obtained in the first separation section 20, and the obtained target particle concentrated liquid Sb is then introduced into the second separation section 30, thereby significantly shortening the time required for processing (collection of target particles) in the second separation section 30, and making it possible to process a large amount of sample liquid in a short time and at high speed. Furthermore, by detecting the liquid level of the liquid in each storage section or recovery section with the liquid level sensors 51-56 and controlling the amount of liquid in each storage section or recovery section to be appropriate, it becomes possible to automate the processing of a large amount of sample liquid without the need for constant monitoring by an operator.

[0119] <Second embodiment> A basic concept according to the second embodiment of the present invention will be described with reference to Fig. 7 and Fig. 8. Fig. 7 is a diagram showing an example of the configuration of a particle separator (CTC separator) according to the second embodiment of the present invention. Fig. 8 is a diagram showing an example of the configuration of a first separation section used in the particle separator (CTC separator) according to the second embodiment of the present invention. In describing Fig. 7, the description of components similar to those in Fig. 1 may be simplified or omitted.

[0120] As shown in Figure 7, the particle separation device 200 in this embodiment is roughly configured to include a sample liquid storage section 211, a first separation section 20a, a target particle concentrated liquid storage section 212, a target particle removal liquid recovery section 213, a second separation section 20b, a second target particle concentrated liquid recovery section 214, and a second target particle removal liquid recovery section 215.

[0121] The sample liquid storage section 211 is a component similar to the sample liquid storage section 11 shown in Fig. 1. The sample liquid storage section 211 stores a sample liquid Sa containing target particles to be separated. An outlet for discharging the sample liquid Sa is connected to one end of the first pipe 241.

[0122] The first separation section 20a has a sample liquid inlet 21a as a liquid inlet, and a target particle concentrated liquid outlet 22a and a target particle removal liquid outlet 23a as liquid outlets. The sample liquid inlet 21a is connected to the other end of a first pipe 241, so that the sample liquid Sa discharged from the outlet of the sample liquid storage section 211 is introduced into the sample liquid inlet 21a through the first pipe 241. The target particle concentrated liquid outlet 22a is connected to one end of a second pipe 242, and the target particle removal liquid outlet 23a is connected to one end of a third pipe 243.

[0123] As shown in Fig. 8, the first separation section 20a is provided with a plurality of concentration tips shown in Fig. 3 and Fig. 4 arranged in parallel. The first separation section 20a shown in Fig. 8 has three concentration tips, but the present invention is not limited to this, and any number of concentration tips can be arranged in parallel. The sample liquid Sa introduced from the sample liquid inlet 21a is distributed to each concentration tip, and is separated into a liquid with an increased concentration of target particles and a liquid from which the target particles have been almost removed in each concentration tip. A target particle concentrated liquid Sb is formed by combining the liquids with increased concentrations of target particles discharged from each concentration tip, and is discharged from a target particle concentrated liquid outlet 22a. A target particle removed liquid Sc is formed by combining the liquids from which the target particles have been almost removed discharged from each concentration tip, and is discharged from a target particle removed liquid outlet 23a.

[0124] The first separation unit 20a is configured to introduce only the sample liquid Sa, and the entire DLD microchannel can be used for the distribution (processing) of the sample liquid Sa, so the processing speed of the sample liquid Sa can be increased compared to a configuration in which two types of liquid (sample liquid Sa and buffer liquid) are introduced. Furthermore, the first separation unit 20a is configured to introduce only the sample liquid Sa, and only the sample liquid Sa needs to be distributed to each chip, so the structure is less likely to become complicated even if the chips are arranged in parallel, compared to a configuration in which two types of liquid (sample liquid Sa and buffer liquid) are introduced, and further, there is no need to adjust the flow rate of the buffer liquid according to the flow rate of the sample liquid Sa. By arranging the chips in parallel, the first separation unit 20a can process a large amount of sample liquid Sa in a short time and at high speed.

[0125] The target particle concentrated liquid storage unit 212 is a component similar to the target particle concentrated liquid storage unit 12 shown in Fig. 1. The other end of the second pipeline 242 is connected to the storage space of the target particle concentrated liquid storage unit 212, and the target particle concentrated liquid Sb discharged from the first separation unit 20a is stored therein. An outlet for discharging the target particle concentrated liquid Sb is connected to one end of the fourth pipeline 244.

[0126] The target particle removal liquid recovery section 213 is a component similar to the target particle removal liquid recovery section 13 shown in Fig. 1. The other end of the third pipe 243 is connected to the storage space of the target particle removal liquid recovery section 213, and the target particle removal liquid Sc discharged from the first separation section 20a is stored therein.

[0127] The second separation section 20b has a target particle concentrated liquid inlet 21b as a liquid inlet, and a second target particle concentrated liquid outlet 22b and a second target particle removal liquid outlet 23b as liquid outlets. The target particle concentrated liquid inlet 21b is connected to the other end of a fourth pipe 244, so that the target particle concentrated liquid Sb discharged from the outlet of the target particle concentrated liquid storage section 212 is introduced into the target particle concentrated liquid inlet 21b through the fourth pipe 244. In addition, the second target particle concentrated liquid outlet 22b is connected to one end of a fifth pipe 245, and the second target particle removal liquid outlet 23b is connected to one end of a sixth pipe 246.

[0128] The second separation section 20b has a function as a concentration chip shown in Fig. 3 and Fig. 4, similar to the first separation section 20 shown in Fig. 1. The second separation section 20b separates the target particle concentrated solution Sb introduced from the target particle concentrated solution inlet 21b into a solution with an increased concentration of target particles and a solution from which the target particles have been almost completely removed. The solution with the increased concentration of target particles is discharged from the second target particle concentrated solution outlet 22b as a second target particle concentrated solution Se, and the solution from which the target particles have been almost completely removed is discharged from the second target particle removed solution outlet 23b as a second target particle removed solution Sf. The second target particle concentrated solution Se is a liquid in which the amount is significantly reduced compared to the target particle concentrated solution Sb, and the target particles are further concentrated.

[0129] The second target particle concentrated liquid recovery section 214 is a container that contains the second target particle concentrated liquid Se discharged from the second separation section 20b. The other end of the fifth pipe 245 is connected to the storage space of the second target particle concentrated liquid recovery section 214, and the second target particle concentrated liquid Se discharged from the second target particle concentrated liquid outlet 22b of the second separation section 20b is introduced into the second target particle concentrated liquid recovery section 214 through the fifth pipe 245. The storage space of the second target particle concentrated liquid recovery section 214 does not need to be airtight, and the air pressure inside the second target particle concentrated liquid recovery section 214 may be equal to atmospheric pressure.

[0130] The second targeted particle removal liquid recovery section 215 is a container that contains the second targeted particle removal liquid Sf discharged from the second separation section 20b. The other end of the sixth pipeline 246 is connected to the storage space of the second targeted particle removal liquid recovery section 215, and the second targeted particle removal liquid Sf discharged from the second targeted particle removal liquid outlet 23b of the second separation section 20b is introduced into the second targeted particle removal liquid recovery section 215 through the sixth pipeline 246. The storage space of the second targeted particle removal liquid recovery section 215 does not need to be airtight, and the air pressure inside the second targeted particle removal liquid recovery section 215 may be equal to atmospheric pressure.

[0131] An example of the operation of the particle separator 200 shown in FIG. 7 will be described.

[0132] First, it is preferable to fill all DLD flow path parts and their liquid introduction pipes (first pipe 241, fourth pipe 244) with liquid (e.g., buffer liquid). After that, the sample liquid Sa is accommodated in the sample liquid accommodation part 211. When the air pressure A in the sample liquid accommodation part 211 is made higher than the air pressure B (e.g., atmospheric pressure) in the target particle concentrated liquid accommodation part 212 and the air pressure B (e.g., atmospheric pressure) in the target particle removed liquid recovery part 213, the sample liquid Sa accommodated in the sample liquid accommodation part 211 is pressure-fed into the first separation part 20a via the first pipe 241. The sample liquid Sa introduced into the first separation part 20a is separated into a liquid with an increased concentration of target particles (target particle concentrated liquid Sb) and a liquid from which the target particles have been almost removed (target particle removed liquid Sc), and then the target particle concentrated liquid Sb is discharged into the target particle concentrated liquid accommodation part 212, and the target particle removed liquid Sc is discharged into the target particle removed liquid recovery part 213.

[0133] In a state where an appropriate amount of the target particle concentrated liquid Sb is stored in the target particle concentrated liquid storage section 212, the air pressure B in the target particle concentrated liquid storage section 212 and the air pressure B in the target particle removal liquid recovery section 213 are made higher than the air pressure (e.g., atmospheric pressure) in the second target particle concentrated liquid recovery section 214 and the air pressure (e.g., atmospheric pressure) in the second target particle removal liquid recovery section 215, so that the target particle concentrated liquid Sb stored in the target particle concentrated liquid storage section 212 is pressure-fed into the second separation section 20b via the fourth pipe 244. The target particle concentrated liquid Sb introduced into the second separation section 20b is separated into a liquid with an increased concentration of target particles (second target particle concentrated liquid Se) and a liquid from which the target particles have been almost removed (second target particle removal liquid Sf), and thereafter the second target particle concentrated liquid Se is discharged into the second target particle concentrated liquid recovery section 214, and the second target particle removal liquid Sf is discharged into the second target particle removal liquid recovery section 215.

[0134] By setting the pressure in the second target particle concentrated liquid recovery section 214 and the pressure in the second target particle removal liquid recovery section 215 to, for example, atmospheric pressure, setting the pressure B in the target particle concentrated liquid storage section 212 and the pressure B in the target particle removal liquid recovery section 213 higher than atmospheric pressure (pressure B>atmospheric pressure), and further setting the pressure A in the sample liquid storage section 211 higher than pressure B (pressure A>pressure B), the liquid in each storage section is continuously pumped in a direction in which it will be subjected to the particle separation action, thereby achieving continuous particle separation. Also, by setting the pressure A and pressure B to atmospheric pressure, the operation of the particle separation device 200 can be interrupted or stopped.

[0135] By making the air pressure inside the target particle concentrated liquid storage unit 212 and the air pressure inside the target particle removal liquid recovery unit 213 equal (air pressure B), the target particle concentrated liquid Sb and the target particle removal liquid Sc are discharged from the first separation unit 20a appropriately without bias. The air pressure inside the target particle concentrated liquid storage unit 212 and the air pressure inside the target particle removal liquid recovery unit 213 may be configured to be equal (air pressure B) by connecting the target particle concentrated liquid storage unit 212 and the target particle removal liquid recovery unit 213 to the same pressure generator. Alternatively, the target particle concentrated liquid storage unit 212 and the target particle removal liquid recovery unit 213 may be configured to be connected to a pipeline through which gas flows to make them equal pressure, and then a pressure generator may be connected to the pipeline to control the air pressure B. Furthermore, the target particle concentrated liquid storage section 212 and the target particle removal liquid recovery section 213 may be connected by a pipeline through which gas flows to make them equal in pressure, and the air pressure B may be indirectly increased by discharging the target particle concentrated liquid Sb from the first separation section 20a into the target particle concentrated liquid storage section 212 and the target particle removal liquid Sc into the target particle removal liquid recovery section 213.

[0136] The particle separation device 200 shown in Fig. 7 is configured to perform two-stage processing, processing in a first separation section 20a and processing in a second separation section 20b. The first separation section 20a can obtain a target particle concentrated liquid Sb from the sample liquid Sa at a high processing speed. The target particle concentrated liquid Sb is a liquid in which the amount of the target particle concentrated liquid Sb is significantly reduced compared to the sample liquid Sa and the target particles are concentrated. After obtaining the target particle concentrated liquid Sb in a significantly reduced amount compared to the sample liquid Sa in the first separation section 20a, the obtained target particle concentrated liquid Sb is introduced into the second separation section 20b, thereby significantly shortening the time required for processing in the second separation section 20b (concentrating and removing the target particles), and making it possible to process a large amount of sample liquid in a short time and at high speed.

[0137] In addition, in this embodiment, a liquid level sensor 51 may be optionally provided to detect the liquid level of the target particle concentrated liquid Sb contained in the target particle concentrated liquid storage section 212, and the target particle concentrated liquid Sb in the target particle concentrated liquid storage section 212 may be maintained at an appropriate amount based on the detection result of the liquid level of the target particle concentrated liquid Sb.

[0138] Next, a variation of this embodiment will be described with reference to Fig. 9. Fig. 9 is a diagram showing a variation of the configuration of the particle separator (CTC separator) in the second embodiment of the present invention. In describing Fig. 9, the description of the same components as those in Figs. 6 and 7 may be simplified or omitted.

[0139] The particle separation device 300 shown in Figure 9 is similar to the particle separation device 200 shown in Figure 7 and is roughly configured to include a sample liquid storage section 211, a first separation section 20a, a target particle concentrated liquid storage section 212, a target particle removal liquid recovery section 213, a second separation section 20b, a second target particle concentrated liquid recovery section 214, and a second target particle removal liquid recovery section 215.

[0140] 9 further includes liquid level sensors 51, 53 to 56, pressure generators 71, 72, a tube pump 74, and a control unit 75, similar to a portion of the configuration of the particle separator 100 shown in FIG.

[0141] The liquid level sensor 51 is similar to that shown in FIG. 7 and has a function of detecting the liquid level of the target particle concentrated liquid Sb contained in the target particle concentrated liquid storage section 212. The liquid level sensor 53 has a function of detecting the liquid level of the sample liquid Sa contained in the sample liquid storage section 211. The liquid level sensor 54 has a function of detecting the liquid level of the target particle removal liquid Sc contained in the target particle removal liquid recovery section 213. The liquid level sensor 55 has a function of detecting the liquid level of the second target particle removal liquid Sf contained in the second target particle removal liquid recovery section 215. The liquid level sensor 56 has a function of detecting the liquid level of the second target particle removal liquid Se contained in the second target particle removal liquid recovery section 214.

[0142] The pressure generator 71 is a device capable of controlling the air pressure (air pressure A) in the sample liquid storage unit 211 to a desired air pressure. The pressure generator 72 is a device capable of controlling the air pressure B in the target particle concentrated liquid storage unit 212 and the air pressure B in the target particle removed liquid recovery unit 213 to be equal and at a desired air pressure.

[0143] The control unit 75 is connected to the liquid level sensors 51, 53 to 56, the pressure generators 71, 72, and the tube pump 74, and is configured to adjust the amount of liquid in each storage unit or recovery unit by controlling the operation of the pressure generators 71, 72, and the tube pump 74 based on the liquid level of the liquid in each storage unit or recovery unit supplied from the liquid level sensors 51, 53 to 56. The control unit 75 is configured by a processor such as a CPU, and can control the pressure generators 71, 72, and the tube pump 74 to perform desired operations by executing program instructions that specify the operation control procedures of the pressure generators 71, 72, and the tube pump 74 according to the liquid level of the liquid in each storage unit or recovery unit.

[0144] The control unit 75 controls the operation of the pressure generators 71, 72 based on the detection result of the liquid level by the liquid level sensor 51, thereby eliminating the need for constant monitoring by an operator and making it possible to maintain an appropriate amount of the target particle concentrated liquid Sb in the target particle concentrated liquid storage unit 212. The control unit 75 controls the operation of the tube pump 74 based on the detection result of the liquid level by the liquid level sensor 53, thereby eliminating the need for constant monitoring by an operator and making it possible to maintain an appropriate amount of the sample liquid Sa in the sample liquid storage unit 211.

[0145] When the liquid level sensor 54 detects that the liquid level of the target particle removal liquid Sc has risen above a predetermined level (upper threshold), the control unit 75 controls a recovery mechanism (not shown) to recover the target particle removal liquid Sc from the target particle removal liquid recovery unit 213. When the liquid level sensor 55 detects that the liquid level of the second target particle removal liquid Sf has risen above a predetermined level (upper threshold), the control unit 75 controls a recovery mechanism (not shown) to recover the second target particle removal liquid Sf from the second target particle removal liquid recovery unit 215. When the liquid level sensor 56 detects that the liquid level of the second target particle concentrated liquid Se has risen above a predetermined level (upper threshold), the control unit 75 controls a recovery mechanism (not shown) to recover the second target particle concentrated liquid Se from the second target particle concentrated liquid recovery unit 214. The control unit 75 may control the operation of the pressure generators 71, 72 to set the atmospheric pressure A and B to atmospheric pressure and temporarily suspend the operation of the particle separation device 300, and then issue an instruction to collect the target particle removal solution Sc, the second target particle removal solution Sf, and the second target particle concentrated solution Se, or may alert the operator by outputting an alarm sound or emitting light, etc., to cause the operator to collect the target particle removal solution Sc, the second target particle removal solution Sf, and the second target particle concentrated solution Se. By monitoring the liquid level detected by the liquid level sensors 54 to 56, it becomes possible to collect the target particle removal solution Sc, the second target particle removal solution Sf, and the second target particle concentrated solution Se at an appropriate timing.

[0146] 9, the liquid level sensors 51, 53 to 56 are provided at a total of five locations, but it is not necessary to provide all of the liquid level sensors 51, 53 to 56, and only one of the liquid level sensors 51, 53 to 56, or a plurality of liquid level sensors arbitrarily selected from the liquid level sensors 51, 53 to 56 may be provided. For example, the liquid level sensors 51 and 53 may be provided, while the liquid level sensors 54 to 56 may not be provided. In this case, when refilling the sample liquid Sa based on the detection result of the liquid level sensor 53, the recovery of the target particle removal liquid Sc, the recovery of the second target particle removal liquid Sf, and the recovery of the second target particle concentrated liquid Se may be performed simultaneously.

[0147] In the particle separator 300 shown in Fig. 9, similarly to the particle separator 200 shown in Fig. 7, a target particle concentrated liquid Sb that is significantly reduced in amount compared to the sample liquid Sa is obtained in the first separation section 20a, and the obtained target particle concentrated liquid Sb is then introduced into the second separation section 20b, thereby significantly shortening the time required for processing (concentration of target particles and removal of target particles) in the second separation section 20b, and it becomes possible to process a large amount of sample liquid in a short time and at high speed. Furthermore, by detecting the liquid level of the liquid in each storage section or recovery section with the liquid level sensors 51, 53 to 56 and controlling the amount of liquid in each storage section or recovery section to be appropriate, it becomes possible to automate the processing of a large amount of sample liquid without the operator having to constantly monitor.

[0148] The particle separation devices 10 and 100 in the first embodiment described above are configured to have a concentration tip in the front stage and a sorting tip in the rear stage, so that the target particles can be extracted into a buffer liquid. In contrast, the particle separation devices 200 and 300 in the second embodiment are configured to have a concentration tip in parallel in the front stage and a concentration tip in the rear stage, so that the target particles cannot be extracted into a buffer liquid, but can be removed from the sample liquid Sa in a shorter time and at a higher speed.

[0149] In the above-mentioned particle separation devices 10 and 100, one concentration chip is provided in the first separation section 20, but a plurality of chips may be provided and arranged in parallel, as in the first separation section 20a of the particle separation devices 200 and 300. In addition, in the particle separation devices 200 and 300, a plurality of concentration chips are provided in the first separation section 20a and arranged in parallel, but a single large concentration chip in which a larger number of DLD microchannels are arranged in parallel may be used, or large concentration chips may be arranged in parallel. The first separation sections 20 and 20a are configured to introduce only the sample liquid Sa, and only the sample liquid Sa needs to be distributed to each DLD microchannel or each chip. Therefore, compared to a configuration in which two types of liquid (sample liquid Sa and buffer liquid) are introduced, the structure is less likely to become complicated even if the DLD microchannels or chips are arranged in parallel, and further, there is no need to adjust the flow rate of the buffer liquid according to the flow rate of the sample liquid Sa. In the first separation section 20, 20a, by appropriately combining the parallelization of the DLD microchannels in the concentration chip and the parallelization of the concentration chip, it becomes possible to process a large amount of sample liquid in a short time at high speed.

[0150] Furthermore, the above-mentioned particle separation devices 10, 100, 200, 300 may be combined with a mechanism for removing blood from the body and then returning it to the body, which is used in a hemodialysis device, an apheresis device, an artificial heart-lung machine, etc., so that liquids (target particle removal solution Sc, second target particle removal solution Sd, Sf) from the blood (sample liquid Sa) removed from the body can be returned to the body, thereby making it possible to separate (remove, separate, concentrate) target particles having a particle size equal to or larger than a certain size from the blood, such as circulating tumor cells.

[0151] The operation of the particle separator according to the present invention will now be described.

[0152] The particle separation devices 10, 100, 200, 300 of the present invention have a first separation section 20, 20a having a DLD microchannel in which a plurality of fine pillars are arranged for circulating a sample liquid Sa containing target particles, and a second separation section 30, 20b connected to the first separation section 20, 20a and having a DLD microchannel in which a plurality of fine pillars are arranged for circulating a target particle concentrated liquid Sb discharged from the first separation section 20, 20a. The first separation section 20, 20a is equipped with a sample liquid inlet 21, 21a into which the sample liquid Sa is introduced, a DLD flow path section 24 having a DLD microchannel structure and circulating the sample liquid Sa introduced from the sample liquid inlet 21, 21a to separate it into a liquid with an increased concentration of target particles and a liquid from which the target particles have been almost removed, target particle concentrated liquid outlets 22, 22a for discharging the liquid with an increased concentration of target particles (target particle concentrated liquid Sb) separated in the DLD flow path section 24, and target particle removed liquid outlets 23, 23a for discharging the liquid from which the target particles have been almost removed (target particle removed liquid Sc) separated in the DLD flow path section 24.

[0153] According to the above configuration, the first separation section 20, 20a separates the sample liquid Sa into a liquid with an increased concentration of target particles (target particle concentrated liquid Sb) and a liquid from which the target particles have been almost completely removed (target particle removed liquid Sc), and discharges the target particle concentrated liquid Sb, which is significantly reduced in volume compared to the sample liquid Sa. The second separation section 30, 20b processes this target particle concentrated liquid Sb, thereby significantly reducing the processing time by the second separation section 30, 20b, and making it possible to process large amounts of sample liquid Sa in a short period of time and at high speed.

[0154] In the particle separation device 10, 100 according to the first embodiment, the second separation section 30 may be provided with a target particle concentrated liquid inlet 31 into which the target particle concentrated liquid Sb discharged from the target particle concentrated liquid outlet 22 of the first separation section 20 is introduced, a buffer liquid inlet 32 ​​into which a buffer liquid Ba is introduced, a DLD flow path section 35 having a DLD microchannel structure and flowing the target particle concentrated liquid Sb introduced from the target particle concentrated liquid inlet 31 and the buffer liquid Ba introduced from the buffer liquid inlet 32 ​​in parallel so that they come into contact with each other, thereby separating the target particle concentrated liquid Sb into a buffer liquid containing the target particles and a liquid from which the target particles have been almost removed, a second target particle removal liquid outlet 33 for discharging the liquid from which the target particles have been almost removed (second target particle removal liquid Sd) separated in the DLD flow path section 35, and a buffer liquid outlet 34 for discharging the buffer liquid containing the target particles separated in the DLD flow path section 35 (recovered buffer liquid Bb).

[0155] According to the above configuration, the second separation section 30 can move the target particles contained in the target particle concentrated solution Sb, which has been significantly reduced in amount compared to the sample solution Sa, to the buffer solution Ba, and separate the solution into a buffer solution containing the target particles (recovery buffer solution Bb) and a solution from which the target particles have been almost completely removed (second target particle removal solution Sd).

[0156] In the particle separation device 200, 300 according to the second embodiment, the second separation section 20b may include a target particle concentrated liquid inlet 21b into which the target particle concentrated liquid Sb discharged from the target particle concentrated liquid outlet 22a of the first separation section 20a is introduced, a DLD flow path section 24 having a DLD microchannel structure and circulating the target particle concentrated liquid Sb introduced from the target particle concentrated liquid inlet 21b to separate the target particle concentrated liquid Sb into a liquid with an increased concentration of target particles and a liquid from which the target particles have been almost removed, a second target particle concentrated liquid outlet 22b for discharging the liquid with the increased concentration of target particles separated in the DLD flow path section 24 (second target particle concentrated liquid Se), and a second target particle removal liquid outlet 23b for discharging the liquid from which the target particles have been almost removed separated in the DLD flow path section 24 (second target particle removal liquid Sf).

[0157] According to the above configuration, the second separation section 20b can separate the target particle concentrated liquid Sb, which has been significantly reduced in volume compared to the sample liquid Sa, into a liquid with an even higher concentration of target particles (second target particle concentrated liquid Se) and a liquid from which the target particles have been almost completely removed (second target particle removed liquid Sf).

[0158] The above-described embodiments are described for the purpose of facilitating understanding of the present invention, and are not described for the purpose of limiting the present invention. Therefore, the elements disclosed in the above-described embodiments are intended to include all design modifications and equivalents that fall within the technical scope of the present invention. [Explanation of symbols]

[0159] 10, 100, 200, 300 particle separation device (CTC separation device) 11, 211 Sample liquid storage section 12, 212 Target particle concentrate storage section 13, 213 Target particle removal liquid recovery section 14 Buffer solution storage section 15, 215 Second target particle removal liquid recovery section 16 Buffer solution recovery section 20, 20a 1st separation section 20b, 30 2nd separation section 21, 21a Sample liquid inlet 21b, 31 Target particle concentrate inlet 22, 22a Target particle concentrate outlet 22b 2nd target particle concentrate outlet 23, 23a Target particle removal liquid outlet 23b, 33 2nd target particle removal liquid outlet 24, 35 DLD flow path section 25 Confluence Channel 32 Buffer solution inlet 34 Buffer solution outlet 41, 241 1st pipeline 42, 242 2nd pipeline 43, 243 3rd pipeline 44, 244 4th pipeline 45, 245 5th pipeline 46, 246 6th pipeline 47 Pipeline 7 51~56 Liquid level sensor 71, 72 Pressure generating device 73 Buffer solution supply unit 74 Tube Pump 75 Control Unit 214 Second target particle concentrated liquid recovery section Sa sample solution Sb target particle concentrate Sc target particle removal solution Sd, Sf 2nd target particle removal solution Se 2nd target particle concentrate Ba buffer solution Bb Recovery buffer

Claims

1. a first separation unit including a DLD microchannel having a plurality of fine pillars arranged therein for flowing a sample liquid containing target particles; a second separation section connected to the first separation section and including a DLD microchannel having a plurality of fine pillars arranged therein for circulating a target particle concentrate discharged from the first separation section; a sample liquid storage section that supplies the sample liquid to the first separation section; a target particle concentrated liquid storage section interposed between the first separation section and the second separation section, storing the target particle concentrated liquid discharged from a target particle concentrated liquid outlet of the first separation section, and supplying the target particle concentrated liquid to the second separation section; a target particle removal liquid recovery section that stores the liquid discharged from a target particle removal liquid discharge port of the first separation section; a buffer solution storage section that supplies a buffer solution to the second separation section; 1. A particle separator comprising: The first separation unit is a sample liquid inlet into which the sample liquid is introduced; a DLD flow path section including a DLD microchannel structure, which circulates the sample liquid introduced from the sample liquid inlet port and separates the sample liquid into a liquid having an increased concentration of the target particles and a liquid from which the target particles have been almost completely removed; a target particle concentrated liquid outlet that discharges a liquid in which the concentration of the target particles separated in the DLD flow path portion has been increased as the target particle concentrated liquid; a target particle removed liquid outlet for discharging a liquid from which the target particles separated in the DLD flow path portion have been almost removed; Equipped with The second separation section, a target particle concentrated liquid inlet into which the target particle concentrated liquid discharged from the target particle concentrated liquid outlet of the first separation unit is introduced; a buffer solution inlet through which the buffer solution is introduced; a DLD flow path section including a DLD microchannel structure, in which the target particle concentrated solution introduced from the target particle concentrated solution inlet and the buffer solution introduced from the buffer solution inlet are allowed to flow in parallel so as to come into contact with each other, thereby separating the target particle-containing buffer solution from a solution from which the target particles have been substantially removed; an outlet for discharging a liquid from which the target particles separated in the DLD flow path portion have been almost completely removed; an outlet for discharging a buffer solution containing the target particles separated in the DLD flow path; Equipped with adjusting the air pressure in the sample liquid storage unit, the air pressure in the target particle concentrated liquid storage unit, the air pressure in the target particle removal liquid recovery unit, and the air pressure in the buffer liquid storage unit simultaneously; A particle separation device characterized in that the air pressure in the sample liquid storage section is made higher than the air pressure in the target particle concentrated liquid storage section, and the air pressure in the target particle concentrated liquid storage section, the air pressure in the target particle removal liquid recovery section, and the air pressure in the buffer liquid storage section are maintained in a state of being approximately equal during particle separation.

2. A liquid level sensor for detecting a liquid level of the buffer liquid in the buffer liquid storage section; a control unit that replenishes the buffer liquid from an external source based on a detection result of the liquid level sensor to adjust the amount of the buffer liquid in the buffer liquid storage unit; 2. The particle separator of claim 1, further comprising:

3. a first separation unit including a DLD microchannel having a plurality of fine pillars arranged therein for flowing a sample liquid containing target particles; a second separation section connected to the first separation section and including a DLD microchannel having a plurality of fine pillars arranged therein for circulating a target particle concentrate discharged from the first separation section; a sample liquid storage section that supplies the sample liquid to the first separation section; a target particle concentrated liquid storage section interposed between the first separation section and the second separation section, storing the target particle concentrated liquid discharged from a target particle concentrated liquid outlet of the first separation section, and supplying the target particle concentrated liquid to the second separation section; a target particle removal liquid recovery section that stores the liquid discharged from a target particle removal liquid discharge port of the first separation section; 1. A particle separator comprising: The first separation unit is a sample liquid inlet into which the sample liquid is introduced; a DLD flow path section including a DLD microchannel structure, which circulates the sample liquid introduced from the sample liquid inlet port and separates the sample liquid into a liquid having an increased concentration of the target particles and a liquid from which the target particles have been almost completely removed; a target particle concentrated liquid outlet that discharges a liquid in which the concentration of the target particles separated in the DLD flow path portion has been increased as the target particle concentrated liquid; a target particle removed liquid outlet for discharging a liquid from which the target particles separated in the DLD flow path portion have been almost removed; Equipped with The second separation section, a target particle concentrated liquid inlet into which the target particle concentrated liquid discharged from the target particle concentrated liquid outlet of the first separation unit is introduced; a DLD flow path section including a DLD microchannel structure, which circulates the target particle concentrated solution introduced from the target particle concentrated solution inlet and separates the target particle concentrated solution into a solution having an increased concentration of the target particles and a solution from which the target particles have been substantially removed; an outlet for discharging a liquid having an increased concentration of the target particles separated in the DLD flow path; an outlet for discharging a liquid from which the target particles separated in the DLD flow path portion have been almost completely removed; Equipped with adjusting the air pressure in the sample liquid storage unit, the air pressure in the target particle concentrated liquid storage unit, and the air pressure in the target particle removal liquid recovery unit simultaneously; A particle separation device characterized in that the air pressure in the sample liquid storage section is made higher than the air pressure in the target particle concentrated liquid storage section, and a state in which the air pressure in the target particle concentrated liquid storage section and the air pressure in the target particle removal liquid recovery section are approximately equal is maintained during particle separation.

4. A particle separation device described in any one of claims 1 to 3, characterized in that the air pressure within the target particle concentrated liquid storage section is higher than atmospheric pressure.

5. A liquid level sensor for detecting a liquid level of the target particle concentrate in the target particle concentrate storage section; a control unit that adjusts the amount of the target particle concentrate in the target particle concentrate storage unit based on a detection result of the liquid level sensor; The particle separator according to any one of claims 1 to 4, further comprising:

6. A liquid level sensor for detecting a liquid level of the sample liquid in the sample liquid storage portion; a control unit that replenishes the sample liquid from the outside based on a detection result of the liquid level sensor, and adjusts the amount of the sample liquid in the sample liquid storage unit; The particle separator according to any one of claims 1 to 5, further comprising:

7. 7. The particle separation device according to claim 1, wherein the sample liquid contains non-target particles having a size different from that of the target particles.

8. The particle separation device according to any one of claims 1 to 7, wherein the target particles are cells.

9. The particle separation device according to claim 8 , wherein the sample liquid contains circulating tumor cells as the target particles.

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