Analysis support device, analysis support system, collection method, and analysis method

Abstract

To provide an analysis support device, an analysis support system, and a recovery method that can recover captured cells.SOLUTION: An analysis support device (2) comprises a base portion (21), an annular frame portion (22), a capturing portion (26), and a lid portion (25). The frame portion (22) protrudes from the base portion (21). The frame portion (22) forms an accommodation space (23) together with the base portion (21). The capturing portion (26) protrudes from the base portion (21) within the accommodation space (23). The capturing portion (26) captures one cell (C). The lid portion (25) opens and closes an open surface (20a) of the accommodation space (23). A height (H1) of the frame portion (22) with respect to the base portion (21) is higher than a height (H2) of the capturing portion (26) with respect to the base portion (21).SELECTED DRAWING: Figure 3

Description

[Technical Field]

[0001] The present invention relates to an analysis support device, an analysis support system, a collection method, and an analysis method. [Background technology]

[0002] Devices that trap cells one by one have been proposed. For example, Non-Patent Document 1 discloses a microfluidic chip that traps cells one by one. The microfluidic chip in Non-Patent Document 1 includes a chamber filled with a cell suspension and multiple trap sites provided in the chamber. By placing the microfluidic chip with the chamber filled with the cell suspension on a spin coater and rotating it in a horizontal plane, one cell is trapped at each trap site. The trapped cells can be observed using a confocal microscope. [Prior art documents] [Non-patent literature]

[0003] [Non-Patent Document 1] Chen Zhu, 8 others, "Single Cell Receptor Analysis Aided by a Centrifugal Microfluidic Device for Immune Cells Profiling", Bulletin of the Chemical Society of Japan, Public Interest Incorporated Association, Chemical Society of Japan, 2019, Vol.92, No.11, p.1834 - 1839 Summary of the Invention [Problem to be solved by the invention]

[0004] However, the microfluidic chip of Non-Patent Document 1 is only capable of observing the cells trapped at the trap site, but is not capable of recovering the cells trapped at the trap site.

[0005] The present invention has been made in view of the above-mentioned problems, and an object of the present invention is to provide an analysis support device, an analysis support system, and a recovery method that are capable of recovering trapped cells. Another object of the present invention is to provide an analysis method for analyzing cells captured by the analysis support device. [Means for solving the problem]

[0006] In the present invention, the analysis support device comprises a base portion, an annular frame portion, a capture portion, and a lid portion. The frame portion protrudes from the base portion. The frame portion forms a storage space together with the base portion. The capture portion protrudes from the base portion within the storage space. The capture portion captures a single cell. The lid portion opens and closes the open side of the storage space. The height of the frame portion relative to the base portion is greater than the height of the capture portion relative to the base portion.

[0007] In one embodiment, the analysis support device further includes a peripheral wall portion. The peripheral wall portion protrudes from the base portion within the storage space. The peripheral wall portion surrounds the capture portion. The peripheral wall portion has at least one gap through which the cell passes. The height of the peripheral wall portion relative to the base portion is greater than the height of the capture portion relative to the base portion and less than the height of the frame portion relative to the base portion.

[0008] In the present invention, an analysis support system includes the above-described analysis support device, a stage, and a support base. The stage rotates around a central axis. The support base is placed on the stage. The support base supports the analysis support device. The support base has an inclined surface on which the analysis support device is placed. The inclined surface is inclined so that the side farther from the central axis is farther from the stage than the side closer to the central axis.

[0009] In the present invention, a recovery method is a method for recovering cells using the above-described analysis support device, and includes the steps of introducing a solution containing the cells into the storage space, closing the open surface of the storage space with the lid, rotating the analysis support device in a tilted state around a central axis distant from the analysis support device to capture one cell in the capture unit, opening the open surface of the storage space, and recovering the cell captured in the capture unit from the storage space.

[0010] In the present invention, an analysis method is a method for analyzing cells using the analysis support device having the peripheral wall portion among the above-mentioned analysis support devices, and includes the steps of introducing a solution containing the cells into the storage space, closing the open face of the storage space with the lid portion, rotating the analysis support device in a tilted state around a central axis distant from the analysis support device to capture one of the cells in the capture portion, opening the open face of the storage space, replacing the solution outside the peripheral wall portion in the storage space with a liquid that separates from the solution, and measuring the phenotype of the cell captured in the capture portion.

[0011] In one embodiment, the above-described analysis method further comprises the step of recovering the cells captured by the capture unit from the storage space after measuring the phenotype of the cells.

[0012] In one embodiment, the above-mentioned analytical method measures a secretion product secreted from the cell in determining the phenotype of the cell. [Effects of the Invention]

[0013] According to the analysis support device, analysis support system, and recovery method of the present invention, it is possible to recover the trapped cells.

[0014] Furthermore, according to the analysis method of the present invention, it is possible to analyze cells captured by the analysis support device. [Brief explanation of the drawings]

[0015] [Figure 1] 1 is a perspective view showing an analysis support device according to an embodiment of the present invention. [Figure 2] 1 is a cross-sectional view of an analysis support device according to an embodiment of the present invention. [Figure 3] 10A and 10B are diagrams showing a procedure in which an operator closes the open surface of the storage space with the lid portion. [Figure 4] 1 is an enlarged cross-sectional view showing a part of an analysis support device according to an embodiment of the present invention. [Figure 5] 1 is an enlarged perspective view showing a part of an analysis support device according to an embodiment of the present invention. [Figure 6] FIG. [Figure 7] FIG. 1 is a plan view of a capture structure. [Figure 8] 1 is a side view showing an analysis support system according to an embodiment of the present invention. [Figure 9] 1 is a perspective view showing an analysis support system according to an embodiment of the present invention. [Figure 10] FIG. 2 is a diagram showing a support stand and an analysis support device. [Figure 11] 10A and 10B are cross-sectional views showing the movement of cells in a rotating analysis support device. [Figure 12] FIG. 10 is a diagram showing the movement of cells outside the peripheral wall. [Figure 13] FIG. 10 is a diagram showing cells captured by a capture unit. [Figure 14] 10A and 10B are diagrams showing the movement of other cells on the outside and inside of the peripheral wall. [Figure 15] 1 is a flowchart showing the steps of an analysis method according to an embodiment of the present invention. [Figure 16] FIG. 1 shows a receiving space filled with a cell suspension. [Figure 17] FIG. 10 is a diagram showing the storage space after the removal operation of the cell suspension. [Figure 18] FIG. 10 shows the storage space after oil has been introduced. [Figure 19] FIG. 1 shows a diagram illustrating a cell recovery process. [Figure 20] FIG. 10 shows a removal operation for removing a cell suspension. [Figure 21] FIG. 10 is a plan view showing a modified example of the capture structure. DETAILED DESCRIPTION OF THE INVENTION

[0016] Hereinafter, embodiments of the analysis support device, analysis support system, recovery method, and analysis method of the present invention will be described with reference to the drawings (FIGS. 1 to 20). However, the present invention is not limited to the following embodiments, and can be implemented in various forms without departing from the spirit of the present invention. Note that where explanations are redundant, they may be omitted as appropriate. Furthermore, in the drawings, the same or equivalent parts are designated by the same reference symbols, and explanations will not be repeated.

[0017] First, an analysis support device 2 of this embodiment will be described with reference to Figures 1 to 7. Figure 1 is a perspective view showing the analysis support device 2 of this embodiment. As shown in Figure 1, the analysis support device 2 includes a device main body 20. The device main body 20 has a base portion 21, a frame portion 22, and a plurality of capture structures 24.

[0018] The base portion 21 is plate-shaped. In this embodiment, the outer shape of the base portion 21 is rectangular. However, the outer shape of the base portion 21 is not limited to a rectangular shape. For example, the outer shape of the base portion 21 may be circular.

[0019] The frame portion 22 is annular. One of the open faces of the frame portion 22 is closed by the base portion 21. The frame portion 22 protrudes from the main surface of the base portion 21 and forms an accommodation space 23 together with the base portion 21. A frame height H1, which is the height of the frame portion 22 relative to the base portion 21, is, for example, 1.5 mm.

[0020] The frame portion 22 is rectangular. More specifically, the frame portion 22 has long sides and short sides. The frame portion length L1, which is the length of the long sides of the frame portion 22 (the length in the longitudinal direction of the frame portion 22), is, for example, 50 mm. The frame portion width L2, which is the length of the short sides of the frame portion 22 (the length in the direction perpendicular to the longitudinal direction of the frame portion 22), is, for example, 20 mm. However, the frame portion 22 is not limited to a rectangular shape. For example, the frame portion 22 may be circular.

[0021] The storage space 23 is the space inside the frame portion 22. A part of the base portion 21 forms the floor surface 23a of the storage space 23. The surface of the storage space 23 opposite the floor surface 23a (base portion 21) is open. Hereinafter, the surface of the storage space 23 opposite the floor surface 23a will be referred to as the "open surface 20a" (see Figure 3). The storage space 23 stores a cell suspension CL. The cell suspension CL is introduced into the storage space 23 from the open surface 20a (see Figure 3). The cell suspension CL is an example of a solution containing cells C.

[0022] The plurality of capture structures 24 are disposed inside the frame 22. Therefore, the plurality of capture structures 24 are disposed within the storage space 23 and are surrounded by the frame 22. More specifically, the plurality of capture structures 24 are provided on the floor surface 23a. Each of the plurality of capture structures 24 captures one cell C within the storage space 23.

[0023] 2 is a cross-sectional view of the analysis support device 2 of this embodiment. As shown in FIG. 2, the analysis support device 2 further includes a lid portion 25. The lid portion 25 opens and closes the open surface 20a (see FIG. 3) of the storage space 23. When the open surface 20a (see FIG. 3) is closed, the lid portion 25 covers the storage space 23. In this embodiment, the lid portion 25 is detachable from the device main body 20, and is supported by the frame portion 22.

[0024] The lid 25 is made of, for example, glass. For example, the lid 25 is a glass slide. Alternatively, the material of the lid 25 may be a synthetic resin such as cycloolefin polymer (COP) or a resin such as polycarbonate resin.

[0025] Next, the lid portion 25 will be described with reference to Fig. 3. Fig. 3 is a diagram showing a procedure in which an operator closes the open surface 20a of the storage space 23 with the lid portion 25. The operator closes the open surface 20a with the lid portion 25 in a state in which the storage space 23 is filled with a liquid (for example, the cell suspension liquid CL).

[0026] Specifically, the worker brings the lid portion 25 into contact with the upper surface 22a of the frame portion 22, and gradually moves the lid portion 25 laterally (from the outside to the inside of the frame portion 22) along the upper surface 22a of the frame portion 22 to cover the open surface 20a of the storage space 23 with the lid portion 25. At this time, due to the interfacial tension of the liquid, part of the liquid in the storage space 23 seeps into the gap between the lid portion 25 and the upper surface 22a of the frame portion 22. Then, due to the interfacial tension of the liquid, the lid portion 25 is held on the upper surface 22a of the frame portion 22.

[0027] According to this embodiment, the lid portion 25 is held to the frame portion 22 by utilizing the interfacial tension of the liquid, and therefore the lid portion 25 is detachable from the frame portion 22. Therefore, the lid portion 25 can open and close the open surface 20a of the storage space 23.

[0028] When opening the open surface 20a, the worker moves the lid portion 25 sideways (from the inside to the outside of the frame portion 22) little by little along the upper surface 22a of the frame portion 22, in the same procedure as when closing the open surface 20a with the lid portion 25, to remove the lid portion 25 from the frame portion 22.

[0029] Next, the analysis support device 2 will be further described with reference to Fig. 4. Fig. 4 is an enlarged cross-sectional view showing a portion of the analysis support device 2 of this embodiment. As shown in Fig. 4, the analysis support device 2 has a structural part 2A and a bottom plate 2B. The bottom plate 2B supports the structural part 2A. Specifically, the structural part 2A is bonded to the bottom plate 2B. For example, the structural part 2A and the bottom plate 2B are bonded by plasma.

[0030] The structural unit 2A has an upper portion of a base portion 21 (see FIG. 2) and a frame portion 22 (see FIG. 2). Therefore, the structural unit 2A has a plurality of capture structures 24 (see FIG. 2). In other words, the plurality of capture structures 24 (see FIG. 2) are formed on the structural unit 2A.

[0031] The structural portion 2A is made of a material capable of forming a fine structure. For example, the structural portion 2A can be made of a synthetic resin such as polydimethylsiloxane (PDMS), a composite of PDMS and glass, or COP. Alternatively, the structural portion 2A can be made of a resin such as polycarbonate resin. The bottom plate 2B can be made of glass or polycarbonate resin.

[0032] Next, the capture structure 24 will be described with reference to Fig. 5. Fig. 5 is an enlarged perspective view of a part of the analysis support device 2 of this embodiment. In detail, Fig. 5 shows an enlarged view of a part of the floor surface 23a.

[0033] 5, the plurality of capture structures 24 are provided on the floor surface 23a. The plurality of capture structures 24 are arranged regularly. In this embodiment, the plurality of capture structures 24 are arranged in a staggered pattern.

[0034] As shown in Figure 5, the capture structure 24 has a capture portion 26 and a peripheral wall portion 27. The capture portion 26 and the peripheral wall portion 27 protrude from the floor surface 23a (base portion 21) within the storage space 23 (see Figure 2). The capture portion 26 captures one cell C. The peripheral wall portion 27 surrounds the capture portion 26.

[0035] Next, the capturing structure 24 will be described with reference to Figures 6 and 7. Figure 6 is a perspective view showing the capturing structure 24. As shown in Figure 6, the capturing portion 26 has a first gap G1. The peripheral wall portion 27 has a second gap G2 and a third gap G3.

[0036] The first gap G1 to the third gap G3 are arranged in a straight line. More specifically, the second gap G2, the first gap G1, and the third gap G3 are arranged in this order. For ease of understanding, the following description will be given by defining the direction in which the first gap G1 to the third gap G3 are arranged as the front-to-rear direction. Furthermore, among the first gap G1 to the third gap G3, the second gap G2 side will be defined as the front side, and the third gap G3 side will be defined as the rear side.

[0037] As shown in FIG. 6, the capture portion 26 has a first protrusion 26a and a second protrusion 26b. The first protrusion 26a and the second protrusion 26b are aligned in a direction perpendicular to the front-rear direction. More specifically, the first protrusion 26a and the second protrusion 26b face each other with a first gap G1 between them. A capture portion height H2, which is the height of the capture portion 26 (first protrusion 26a and second protrusion 26b) relative to the floor surface 23a (base portion 21), is lower than the frame height H1 described with reference to FIG. 1. For example, the capture portion height H2 is 25 μm.

[0038] As shown in FIG. 6, the peripheral wall portion 27 forms a trapping space TS together with the base portion 21 (floor surface 23a). The trapping space TS is the space inside the peripheral wall portion 27. A part of the floor surface 23a of the storage space 23 forms the floor surface of the trapping space TS. The surface of the trapping space TS opposite the floor surface 23a is open. The trapping space TS stores a cell suspension CL. The cell suspension CL is held in the trapping space TS.

[0039] The peripheral wall portion 27 has a first peripheral wall portion 27a and a second peripheral wall portion 27b. The first peripheral wall portion 27a and the second peripheral wall portion 27b are aligned in a direction perpendicular to the front-rear direction. More specifically, the first peripheral wall portion 27a and the second peripheral wall portion 27b face each other with a second gap G2 and a third gap G3 in between.

[0040] Specifically, the first circumferential wall portion 27a and the second circumferential wall portion 27b each form a part of a cylindrical shape. Therefore, the first circumferential wall portion 27a and the second circumferential wall portion 27b are arc-shaped in a plan view (see FIG. 7). Front end portions 271 of the first circumferential wall portion 27a and the second circumferential wall portion 27b face each other across a second gap G2. Rear end portions 272 of the first circumferential wall portion 27a and the second circumferential wall portion 27b face each other across a third gap G3.

[0041] The inner diameter of the peripheral wall 27 is, for example, 70 μm. The peripheral wall height H3, which is the height of the peripheral wall 27 (first peripheral wall 27a and second peripheral wall 27b) relative to the floor surface 23a (base portion 21), is higher than the capture portion height H2 and lower than the frame portion height H1 described with reference to Figure 1. The peripheral wall height H3 is, for example, 100 μm.

[0042] Next, the first gap G1 to the third gap G3 will be described with reference to Fig. 7. Fig. 7 is a plan view of the capturing structure 24. The second gap G2 and the third gap G3 have widths W2 and W3, respectively, that satisfy the following two conditions.

[0043] The first condition is that at least one cell C to be captured can pass through the second gap G2 and the third gap G3. The second condition is that the cell suspension CL contained in the capture space TS does not leak out from the second gap G2 and the third gap G3. In other words, the second condition is that the cell suspension CL is held in the capture space TS by the surface tension of the cell suspension CL. The width W2 of the second gap G2 and the width W3 of the third gap G3 may be, for example, two or three times the size of the cell C to be captured. The width W2 of the second gap G2 and the width W3 of the third gap G3 are, for example, 20 μm.

[0044] The capturing unit 26 captures one cell C in the first gap G1. Specifically, the cell C fits into the first gap G1, causing the capturing unit 26 to capture the cell C. Therefore, the width W1 of the first gap G1 is smaller than the dimensions of the cell C to be captured. For example, the width W1 of the first gap G1 is 5 μm.

[0045] Next, the capturing portion 26 will be described with reference to Fig. 7. As shown in Fig. 7, the capturing portion 26 is located at the center of the capturing space TS in a plan view. The first protrusion 26a and the second protrusion 26b extend in a fan-shaped manner from the first gap G1 toward the rear side (the third gap G3 side).

[0046] Specifically, the tip ends 261 (front ends) of the first protrusion 26a and the second protrusion 26b face each other with a first gap G1 in between. The first protrusion 26a becomes farther from the second protrusion 26b as it moves away from the first gap G1 toward the rear (third gap G3 side). Similarly, the second protrusion 26b becomes farther from the first protrusion 26a as it moves away from the first gap G1 toward the rear (third gap G3 side). Therefore, the gap between the respective distal ends 262 (rear ends) of the first protrusion 26a and the second protrusion 26b is larger than the first gap G1.

[0047] Next, an analysis support system 100 and an analysis support device 2 of this embodiment will be described with reference to Figures 1 to 14. First, the analysis support system 100 of this embodiment will be described with reference to Figures 8 and 9. Figure 8 is a side view showing the analysis support system 100 of this embodiment. Figure 9 is a perspective view showing the analysis support system 100 of this embodiment.

[0048] 8 and 9, the analysis support system 100 includes the analysis support device 2 described with reference to FIGS. 1 to 7, a stage 3, two support bases 4, and a rotation mechanism 5 (see FIG. 8). Note that the rotation mechanism 5 is omitted in FIG. 9 to simplify the drawing.

[0049] The stage 3 is rotatable around a central axis AX. In this embodiment, the stage 3 is disk-shaped. The diameter of the stage 3 is, for example, 200 mm. However, the shape of the stage 3 is not limited to a disk-shape. The stage 3 may have any shape as long as it can accommodate at least one support base 4.

[0050] Each of the two support stands 4 is capable of supporting an analysis support device 2. Each support stand 4 is placed on the upper surface of the stage 3. The analysis support device 2 supported by the support stand 4 is held at a position away from the central axis AX. The length L3 (see FIG. 9) of each support stand 4 is, for example, 80 mm.

[0051] The two support bases 4 are disposed at positions where the stage 3 can rotate while maintaining a substantially horizontal state. In this embodiment, the two support bases 4 face each other across the central axis AX. However, the positions at which the two support bases 4 are disposed are not limited to positions where the stage 3 can rotate while maintaining a substantially horizontal state. Other members may be provided on the stage 3 so that the stage 3 can rotate while maintaining a substantially horizontal state.

[0052] The two support stages 4 each have an inclined surface 41 on which the analysis support device 2 is placed. The inclined surface 41 is inclined so that the side farther from the central axis AX is farther from the stage 3 than the side closer to the central axis AX. Therefore, when the analysis support device 2 is placed on the inclined surface 41, the side farther from the central axis AX is farther from the stage 3 than the side closer to the central axis AX.

[0053] The inclination angle θ of the inclined surface 41 with respect to the upper surface of the stage 3 (see FIG. 8) is, for example, greater than 0° and equal to or less than 10°. However, the inclination angle θ of the inclined surface 41 may be any angle that allows the cell C to move along the floor surface 23a described with reference to FIGS. 1 to 7 when the stage 3 is rotated (when the analysis support device 2 is rotated).

[0054] 6 and 7 are aligned (front-rear direction) and the direction in which the inclined surface 41 extends (radial direction of the stage 3). More specifically, the analysis support device 2 is positioned so that its front side is on the central axis AX side (lower end side of the support base 4 and inclined surface 41) and its rear side is on the peripheral edge side of the stage 3 (upper end side of the support base 4 and inclined surface 41). The analysis support device 2 is positioned on the inclined surface 41 of the support base 4 with the storage space 23 filled with the cell suspension CL and the open surface 20a of the storage space 23 closed by the lid portion 25.

[0055] The rotation mechanism 5 (see FIG. 8) rotates the stage 3 about the central axis AX. As a result, the analysis support device 2, which is disposed at a position away from the central axis AX, turns (rotates) about the central axis AX. In other words, the analysis support device 2 turns (rotates) around the central axis AX. As the analysis support device 2 rotates, centrifugal force acts on the cells C in the cell suspension CL, causing the cells C to move toward the rear side of the storage space 23 (the upper end side of the inclined surface 41).

[0056] The rotation speed of the rotation mechanism 5 can be set to any value. For example, the rotation speed of the rotation mechanism 5 is 1000 rpm. The rotation speed of the rotation mechanism 5 is set within a range that does not impose an excessive load on the cells C. Here, an excessive load refers to a load that causes the cells C to become abnormal.

[0057] The rotation mechanism 5 has a rotation shaft 51 and a drive unit 52. The rotation shaft 51 extends along a central axis AX. One end of the rotation shaft 51 is coupled to the stage 3. The drive unit 52 generates a rotational force to rotate the rotation shaft 51 about the central axis AX. As a result, the stage 3 rotates. The drive unit 52 includes, for example, a motor.

[0058] Next, the support base 4 will be described with reference to Fig. 10. Fig. 10 is a diagram showing the support base 4 and the analysis support device 2. In detail, Fig. 10 shows the analysis support device 2 placed on an inclined surface 41 of the support base 4. As shown in Fig. 10, the support base 4 has a support portion 42 that detachably supports the analysis support device 2. In this embodiment, the support portion 42 is provided on the inclined surface 41.

[0059] More specifically, the support portion 42 includes a plurality of protrusions 421. The plurality of protrusions 421 each protrude from the inclined surface 41. The plurality of protrusions 421 are arranged so as to surround the analysis support device 2. More specifically, the plurality of protrusions 421 are arranged so as to contact the edge of the analysis support device 2. The plurality of protrusions 421 surround the analysis support device 2, so that the analysis support device 2 is supported by the inclined surface 41 and the plurality of protrusions 421. Furthermore, by using the plurality of protrusions 421, the analysis support device 2 can be detachably arranged on the inclined surface 41.

[0060] Next, the analysis support device 2 and analysis support system 100 of this embodiment will be described with reference to Figs. 11 to 14. Fig. 11 is a cross-sectional view showing the movement of a cell C in the analysis support device 2 during rotation. Fig. 12 is a diagram showing the movement of a cell C outside the peripheral wall portion 27. Fig. 13 is a diagram showing a cell C captured by the capture portion 26. Fig. 14 is a diagram showing the movement of another cell C outside and inside the peripheral wall portion 27.

[0061] 8 and 9, the analysis support device 2 rotates (pivots) around the central axis AX. As a result, centrifugal force toward the radially outward direction of the stage 3, as described with reference to Figs. 8 and 9, acts on the cells C in the cell suspension CL contained in the containing space 23 of the analysis support device 2, causing the cells C to move toward the radially outward direction of the stage 3.

[0062] Specifically, because the analysis support device 2 is inclined, when centrifugal force acts on the cell C, the cell C moves toward the floor surface 23a, as shown in FIG. 11. Then, after the cell C approaches the floor surface 23a, it moves (rises) along the floor surface 23a. In other words, the cell C moves along the inclined surface 41 of the support base 4. Therefore, the cell C moves from the front to the rear in the front-to-rear direction. As a result, the cell C enters the trapping space TS through the second gap G2 of the peripheral wall portion 27, as shown in FIG. 12, and is trapped in the first gap G1 of the trapping portion 26, as shown in FIG. 13.

[0063] According to this embodiment, the analysis support device 2 can be rotated in an inclined state. Therefore, the cell C can be moved along the floor surface 23a. Therefore, even if the capturing portion height H2 (see FIG. 6) is lower than the frame portion height H1 (see FIG. 1), the cell C can be captured by the capturing portion 26.

[0064] 14, cells C moving at a position outside the second gap G2 of the peripheral wall portion 27 move from the front side to the rear side along the outer circumferential surface of the peripheral wall portion 27. According to this embodiment, the first peripheral wall portion 27a and the second peripheral wall portion 27b are arc-shaped in plan view, and therefore the movement of cells C is not hindered by the peripheral wall portion 27.

[0065] 14, after one cell C is captured by the capturing part 26, the cell C that enters the capturing space TS moves from the front side to the rear side along the outer surface of the capturing part 26 (first protrusion 26a or second protrusion 26b). According to this embodiment, as described with reference to FIG. 7, the capturing part 26 (first protrusion 26a and second protrusion 26b) extends in a fan-shaped manner from the first gap G1 toward the rear side (third gap G3), so that the movement of the cell C is not hindered by the capturing part 26 (first protrusion 26a and second protrusion 26b).

[0066] Next, the recovery method and analysis method of this embodiment will be described with reference to Figs. 1 to 19. Fig. 15 is a flowchart showing the steps of the analysis method of this embodiment. As shown in Fig. 15, the analysis method of this embodiment includes steps S1 to S10. Fig. 16 is a diagram showing the storage space 23 filled with cell suspension CL. Fig. 17 is a diagram showing the storage space 23 after a removal operation for removing the cell suspension CL. Fig. 18 is a diagram showing the storage space 23 after oil M has been introduced. Fig. 19 is a diagram showing a recovery operation for cells C. The analysis method of this embodiment includes a recovery method.

[0067] 15, the operator first introduces the cell suspension CL into the storage space 23 of the analysis support device 2 (step S1). As a result, the cell suspension CL is stored in the storage space 23 as shown in FIG.

[0068] Specifically, the operator drips the cell suspension CL from the open surface 20a into the storage space 23, causing the cells C to settle to the floor surface 23a. At this time, the operator drips the cell suspension CL from the open surface 20a into the storage space 23 until the storage space 23 is filled with the cell suspension CL up to the open surface 20a.

[0069] After introducing the cell suspension CL into the storage space 23, the operator closes the open surface 20a of the storage space 23 with the lid portion 25 as described with reference to FIG. 3 (step S2).

[0070] After closing the open surface 20a of the storage space 23 with the lid 25, the worker places the analysis support device 2 on the inclined surface 41 of the support stand 4 (step S3).

[0071] After placing the analysis support device 2 on the inclined surface 41 of the support base 4, the operator rotates the stage 3 to rotate (pivot) the analysis support device 2 around the central axis AX (step S4). At this time, the analysis support device 2 rotates in an inclined state. As a result, as described with reference to FIGS. 11 to 14, one cell C is captured in each capture section 26. The rotation time is, for example, one minute.

[0072] After stopping the rotation of the stage 3, the operator removes the analysis support device 2 from the inclined surface 41 of the support base 4 (step S5).

[0073] After removing the analysis support device 2 from the support stand 4, the worker removes the lid portion 25 from the device body 20 as described with reference to FIG. 3 to open the open surface 20a of the storage space 23 (step S6).

[0074] After opening the open surface 20a of the storage space 23, the operator replaces the cell suspension liquid CL outside each capture structure 24 (each peripheral wall portion 27) with oil M (step S7). The oil M is an example of a liquid that separates from the cell suspension liquid CL. In other words, the oil M is an example of a liquid that does not mix with the cell suspension liquid CL. The oil M is, for example, mineral oil.

[0075] Specifically, the operator first removes the cell suspension CL outside each capture structure 24 (each peripheral wall portion 27) from the storage space 23. Specifically, the cell suspension CL is absorbed using an absorbent material made of a water-absorbent material such as cotton, paper, or a high molecular weight polymer.

[0076] At this time, as explained with reference to Figures 6 and 7, the cell suspension CL in each trapping space TS is retained in each trapping space TS. Therefore, as shown in Figure 17, in the storage space 23, the cell suspension CL outside each trapping structure 24 (each peripheral wall portion 27) is absorbed and removed from the storage space 23, while the cell suspension CL in each trapping space TS remains (remains) in each trapping space TS. As a result, the cell suspension CL remaining in each trapping space TS forms a droplet separated from the surroundings.

[0077] After the water absorption operation, the worker introduces oil M into the storage space 23. More specifically, the worker drips oil M into the storage space 23 from the open surface 20a. According to this embodiment, as shown in FIG. 18, by introducing oil M into the storage space 23, each droplet of cell suspension CL (cell suspension CL in each trapping space TS) is surrounded by oil M. As described with reference to FIG. 6, since the peripheral wall height H3 is lower than the frame height H1, the upper side of the cell suspension CL in each trapping space TS is also surrounded by oil M.

[0078] After introducing the oil M, the operator measures the phenotype of the cells C captured in each capture section 26 (step S8). For example, the operator measures secretions secreted from each cell C. The secretions are, for example, cytokines or granzymes. The secretions can be observed using a fluorescence microscope such as a confocal fluorescence microscope.

[0079] After measuring the phenotype of each cell C, the operator collects the cell C that has released the target secretory product from the capture unit 26 (step S9). For example, as shown in FIG. 19, the operator can collect the cell C using a micromanipulator K. The target secretory product is, for example, a cytokine or a granzyme.

[0080] The operator analyzes the collected cells C (step S10). For example, the operator may analyze the genetic information of the cells C or the proteome of the cells C.

[0081] Next, with reference to Fig. 20, a removal operation for removing the cell suspension CL outside each capture structure 24 (each peripheral wall portion 27) from the storage space 23 will be described. Fig. 20 is a diagram showing the removal operation for removing the cell suspension CL.

[0082] When the cell suspension CL is absorbed using an absorbent material as described with reference to Fig. 15, the cell suspension CL may remain in places other than the capture space TS of the storage space 23. In this case, after the absorption work, the operator may further remove the cell suspension CL from the storage space 23 using a gas emitter AD such as an air duster, as shown in Fig. 20.

[0083] 6, in this embodiment, the peripheral wall height H3 is higher than the capture part height H2, so the cells C captured by the capture part 26 are not exposed above the liquid surface of the cell suspension CL held in the capture space TS. Therefore, the cells C are unlikely to be blown away from the capture part 26 by the gas ejected from the gas emitter AD.

[0084] As described above with reference to Figures 1 to 20, according to this embodiment, the open surface 20a of the storage space 23 can be opened and closed using the lid portion 25, so that the cells C captured by each capture portion 26 can be recovered one by one.

[0085] Furthermore, according to this embodiment, because the frame height H1 is higher than the capture part height H2, cells C are less likely to come off from the capture part 26 when the storage space 23 is opened. Specifically, when the lid 25 is removed from the frame 22, the liquid surface of the cell suspension CL in the storage space 23 may sway. If the frame height H1 and the capture part height H2 are equal, the swaying of the liquid surface of the cell suspension CL may act on the cells C captured in the capture part 26, causing the cells C to come off from the capture part 26. In contrast, according to this embodiment, because the frame height H1 is higher than the capture part height H2, the swaying of the liquid surface of the cell suspension CL is less likely to act on the cells C captured in the capture part 26. Therefore, cells C are less likely to come off from the capture part 26 when the storage space 23 is opened.

[0086] Furthermore, according to this embodiment, after measuring the phenotype of the cells C captured by each capture unit 26, the cells C can be collected one by one. Therefore, after detecting cells C having a desired phenotype in the analysis support device 2, the cells C having the desired phenotype can be collected and the genetic information of the cells C can be analyzed, or the cytokines or granzymes secreted by the cells C can be analyzed. This makes it possible to improve the efficiency of cell analysis.

[0087] Furthermore, according to this embodiment, the open surface 20a of the storage space 23 can be closed with the lid 25, so that the cells C in the cell suspension CL can be moved by rotating the analysis support device 2. Therefore, one cell C can be captured by each capture part 26.

[0088] Furthermore, according to this embodiment, the phenotype of the cells C trapped in each trapping space TS can be measured while the cell suspension CL held in each trapping space TS is separated by oil M. Therefore, the phenotype of the cells C can be measured in an environment where the secretion from one cell C does not mix with the secretion from another cell C. This improves the accuracy of measuring the phenotype of the cells C.

[0089] Furthermore, according to this embodiment, the cell suspension CL held in each trapping space TS can be surrounded by oil M. Therefore, the cell suspension CL held in each trapping space TS is less likely to evaporate. This makes it easier to measure the phenotype of the cells C. In other words, if the cell suspension CL is not surrounded by oil M, the cell suspension CL is more likely to evaporate. Therefore, it is necessary to finish measuring the phenotype of the cells C before the cell suspension CL evaporates. In contrast, according to this embodiment, the cell suspension CL is less likely to evaporate, so measurement can be performed without worrying about the time required to measure the phenotype of the cells C.

[0090] The embodiments of the present invention have been described above with reference to the drawings (FIGS. 1 to 20). However, the present invention is not limited to the above embodiments and can be implemented in various forms without departing from the spirit of the present invention. Furthermore, the components disclosed in the above embodiments can be modified as appropriate. For example, some of the components shown in one embodiment may be added to the components of another embodiment, or some of the components shown in one embodiment may be deleted from the embodiment.

[0091] The drawings mainly show each component in a schematic manner to facilitate understanding of the invention, and the thickness, length, number, spacing, etc. of each component shown in the drawings may differ from the actual ones due to the convenience of creating the drawings. Furthermore, the configuration of each component shown in the above embodiment is merely an example and is not particularly limited, and it goes without saying that various modifications are possible within a range that does not substantially deviate from the effects of the present invention.

[0092] For example, in the embodiment described with reference to FIGS. 1 to 20, the analysis support system 100 includes two support stages 4, but the number of support stages 4 may be one, or three or more.

[0093] 1 to 20, the analysis support device 2 includes the peripheral wall portion 27, but the peripheral wall portion 27 may be omitted. For example, when the analysis support device 2 is used only for collecting the cells C, the peripheral wall portion 27 can be omitted.

[0094] 1 to 20, the peripheral wall 27 is generally circular in plan view, but the shape of the peripheral wall 27 is not particularly limited as long as the movement of the cells C is not inhibited by the peripheral wall 27. For example, the peripheral wall 27 may be generally hexagonal or generally elliptical in plan view.

[0095] 1 to 20, the analysis support device 2 is configured using two members (the structural portion 2A and the bottom plate 2B), but the analysis support device 2 may also be configured using a single member. In this case, the material for the analysis support device 2 can be, for example, PDMS, a composite of PDMS and glass, or a synthetic resin such as COP. Alternatively, the material for the analysis support device 2 can be a resin such as polycarbonate resin.

[0096] Furthermore, in the embodiment described with reference to Figures 1 to 20, the lid portion 25 is held to the frame portion 22 by the interfacial tension of the liquid, but the device body 20 and the lid portion 25 may have a structure in which the lid portion 25 is removably engaged with the device body 20.

[0097] 1 to 20, lid portion 25 is detachable from device body 20, but lid portion 25 may be connected to device body 20 so as to be openable and closable. For example, device body 20 may have a hinge mechanism for opening and closing lid portion 25.

[0098] 1 to 20, the capturing unit 26 captured one cell C using the first gap G1, but the structure for capturing one cell C is not limited to a gap. A modified example of the capturing unit 26 will be described below with reference to Fig. 21. Fig. 21 is a plan view showing a modified example of the capturing structure 24.

[0099] 21, the capture structure 24 has a capture portion 26 and a peripheral wall portion 27. In the modification shown in Fig. 21, the first protrusion 26a and the second protrusion 26b of the capture portion 26 are connected, and the capture portion 26 has a recess 263 instead of the first gap G1 described with reference to Figs.

[0100] The recess 263 is provided at the connection point between the first protrusion 26a and the second protrusion 26b. The recess 263 is aligned in a straight line with the second gap G2 and the third gap G3 in the front-to-rear direction. The recess 263 is recessed from the front side to the rear side. The recess 263 has a size that allows one cell C to fit therein.

[0101] 21, similar to the capture structure 24 described with reference to FIGS. 1 to 20, the capture portion 26 can capture one cell C. Specifically, the capture portion 26 captures one cell C in the recess 263. [Industrial Applicability]

[0102] The present invention is useful in the field of cell analysis, for example, in the medical field. [Explanation of symbols]

[0103] 2: Analysis support device 3: Stage 4: Support stand 20a: Open surface 21: Base part 22: Frame 23: Containment space 25: Lid 26: Capture unit 27: Peripheral wall part 41: Inclined surface 100: Analysis support system AX: Central axis C: Cell CL: Cell suspension G1: First gap G2: Second gap G3: Third gap H1: Frame height H2: Height of the capture part H3: Peripheral wall height M: Oil

Claims

1. A base portion; an annular frame portion protruding from the base portion and forming an accommodation space together with the base portion; a capture portion that protrudes from the base portion within the storage space and captures one cell; a lid portion that opens and closes the open surface of the storage space; Equipped with a height of the frame portion relative to the base portion being greater than a height of the capture portion relative to the base portion; a peripheral wall portion that protrudes from the base portion within the accommodation space and surrounds the capture portion; the peripheral wall portion has at least one gap through which the cells pass, a height of the peripheral wall portion relative to the base portion is higher than a height of the capturing portion relative to the base portion and is lower than a height of the frame portion relative to the base portion; a height of the peripheral wall portion relative to the base portion is greater than a height of the cells relative to the base portion; the capturing portion has a first gap that captures the cell, the peripheral wall portion has a second gap and a third gap as the gap, The analysis support device, wherein the first gap, the second gap, and the third gap are aligned in a straight line.

2. The analysis support device according to claim 1 , wherein the lid portion moves along an upper surface of the frame portion.

3. The analysis support device according to claim 1 or 2; a stage that rotates around a central axis; a support table disposed on the stage and supporting the analysis support device; Equipped with the support base has an inclined surface on which the analysis support device is placed, The inclined surface is inclined so that the side farther from the central axis is farther from the stage than the side closer to the central axis.

4. A method for recovering cells using the analysis support device according to claim 1 or 2, comprising: introducing a solution containing the cells into the storage space; a step of closing an open surface of the storage space with the lid portion; a step of rotating the analysis support device in a tilted state around a central axis distant from the analysis support device to capture one of the cells in the capture unit; opening the open surface of the storage space; a step of recovering the cells captured in the capture unit from the storage space; a recovery method, including

5. A method for analyzing cells using the analysis support device according to claim 1, introducing a solution containing the cells into the storage space; a step of closing an open surface of the storage space with the lid portion; a step of rotating the analysis support device in a tilted state around a central axis distant from the analysis support device to capture one of the cells in the capture unit; opening the open surface of the storage space; replacing the solution outside the peripheral wall portion in the storage space with a liquid that is separate from the solution; measuring the phenotype of the cells captured in the capture section; Analytical methods, including:

6. The analytical method according to claim 5 , further comprising the step of recovering the cells captured by the capture unit from the storage space after measuring the phenotype of the cells.

7. The analytical method according to claim 5 or 6, wherein a secretion secreted from the cells is measured in measuring the phenotype of the cells.

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