Collection and placement member, collection and placement device, and collection and placement method

Abstract

To provide a collection and placement member, a collection and placement device, and a collection and placement method capable of collecting and placing a sample so as to have high stability, reproducibility, and positional accuracy.SOLUTION: A collection and placement member comprises: a hollow collection pin 141 that can cut out and collect a material to be processed; a placement pin 142 that contacts the material to be processed inside the collection pin 141, in the collection pin 141 and presses the material to be processed to place the material to be processed outside the collection pin 141; and a fixing holder 143 in which the collection pin 141 is fixed. The material to be processed is a gel sheet in which a gel that supports an object to be collected is formed into a sheet shape.SELECTED DRAWING: Figure 2

Description

[Technical Field]

[0001] The present invention relates to a sampling and placement member, a sampling and placement device, and a sampling and placement method. [Background technology]

[0002] In recent years, techniques for selecting cells and cell populations of interest through single-cell analysis have been rapidly developing. Cell selection methods such as limiting dilution, cell sorters, microfluidics, and colony picking have attracted attention.

[0003] Furthermore, printing techniques that handle small amounts of droplets are attracting attention as a technique for handling small amounts of cells. Printing techniques include inkjet printing, dispenser printing, and coating printing.

[0004] However, there are cases where it is preferable to collect only a desired number of desired cells. From the viewpoint of appropriately collecting only a desired number of desired cells, a technique for extracting even smaller amounts of substances with higher precision than the above-mentioned printing technique is required. From this viewpoint, a method for cutting and collecting desired cells from a biological specimen or a cell group using a collection needle disclosed in International Publication No. 2017 / 061387 (Patent Document 1) has been developed.

[0005] Furthermore, as a method for applying minute droplets, Japanese Patent Laid-Open No. 2020-015016 (Patent Document 2) discloses a technology that uses a coating needle whose outer periphery is surrounded by an outer cylinder, and forms droplets that protrude outward from the tip of the coating needle by moving the outer cylinder relative to the coating needle. According to Japanese Patent Laid-Open No. 2020-015016, the liquid material can be applied stably. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] International Publication No. 2017 / 061387 [Patent Document 2] Japanese Patent Publication No. 2020-015016 Summary of the Invention [Problem to be solved by the invention]

[0007] In International Publication No. 2017 / 061387, silicone rubber and gel come into contact with the cell sample. A cut cell sample is placed inside a collection needle. Liquid is then injected into the collection needle, and the resulting pressure expels the cell sample from the needle. However, the silicone rubber and gel do not support and secure the tissue and cells. Therefore, the cell sample crumbles and dissipates when expelled from the collection needle. This can reduce the accuracy of sample collection and placement.

[0008] In addition, in JP 2020-015016 A, the outer cylinder is connected to a spring. Therefore, if an external vertical force is applied to the outer cylinder, it is difficult to maintain the fixed sliding position of the outer cylinder. In other words, the mechanism of JP 2020-015016 A can cause the outer cylinder to move unintentionally when pressed against the sample, making it difficult to cut out the sample.

[0009] The present invention has been made in consideration of the above-mentioned problems, and an object of the present disclosure is to provide a sampling and placement member, a sampling and placement device, and a sampling and placement method that are capable of sampling and placing a sample with high stability, reproducibility, and positional accuracy. [Means for solving the problem]

[0010] The collection and placement member according to the present disclosure includes a collection pin, a placement pin, and a fixing holder. The collection pin is hollow and can cut out and collect the material to be processed. The placement pin can be inserted into the collection pin, and when pressed by the collection pin, it comes into contact with the cut material to be processed inside the collection pin and presses the material to be processed, thereby placing the material to be processed outside the collection pin. The collection pin is fixed to the fixing holder. The material to be processed is a gel sheet formed into a sheet-like gel that supports the object to be collected.

[0011] The sampling and placement device according to the present disclosure includes the sampling and placement member described above and a holding table that holds the material to be treated that is cut out by the sampling pin.

[0012] In the collection and placement method according to the present disclosure, the collection and placement member is first prepared. By lowering the tip of the collection pin, the material to be treated located below the collection pin is cut and placed into the collection pin, and as the tip of the placement pin lowers, it contacts and presses the material to be treated inside the collection pin, thereby placing the material outside the collection pin.

[0013] In the collection and placement method according to the present disclosure, a gel supporting the object to be collected is formed into a sheet on a film, and a gel sheet is provided. The gel sheet on the film is punctured with a hollow collection pin, and a portion of the gel sheet is cut off, and the portion of the gel sheet collected by the collection pin is pushed out from the collection pin by the pressure of the placement pin and placed outside the collection pin. In the above-mentioned placement step, the placement pin is inserted into the collection pin. [Effects of the Invention]

[0014] According to the present disclosure, it is possible to provide a sampling and placement member, a sampling and placement device, and a sampling and placement method that are capable of sampling and placing samples with high stability, reproducibility, and positional accuracy. [Brief explanation of the drawings]

[0015] [Figure 1]1 is a schematic front view of the inside of a collection and placement device according to an embodiment of the present invention. [Figure 2] 2 is a front view of the collection and placement mechanism constituting the collection and placement member of FIG. 1, viewed from the negative side in the Y direction. [Figure 3] 3 is a side view of the collection and placement mechanism of FIG. 2 as viewed from the negative side in the X direction. [Figure 4] 2 is a front view of the entire collection and placement member of FIG. 1, viewed from the negative side in the Y direction. [Figure 5] 5 is a side view of the collection and placement member of FIG. 4 as viewed from the negative side in the X direction. [Figure 6] 5 is a front view of the collection and placement member of FIG. 4, seen from the negative side in the Y direction, showing the operation of the process of cutting and collecting the material to be treated. [Figure 7] 7 is a side view of the operation of FIG. 6 as viewed from the negative side in the X direction. [Figure 8] 3 is a front view of the collection and placement mechanism of FIG. 2, showing the operation of a first step of releasing and placing the material to be treated, as viewed from the negative side in the Y direction. [Figure 9] 9 is a front view showing the operation in the second step following FIG. 8, as viewed from the negative side in the Y direction. [Figure 10] 10 is a front view showing the operation in the third step following FIG. 9, as viewed from the negative side in the Y direction. [Figure 11] 11 is a side view of the operation of FIG. 10 as viewed from the negative side in the X direction. [Figure 12] 11 is a front view of the operation in the fourth step following FIG. 10, as viewed from the negative side in the Y direction. [Figure 13] 13 is a front view showing the operation in the fifth step following FIG. 12, as viewed from the negative side in the Y direction. [Figure 14] 1 is a schematic cross-sectional view showing a first step of the collection and placement method according to the present embodiment. FIG. [Figure 15] 10 is a schematic cross-sectional view showing a second step of the collection and placement method according to the present embodiment. FIG. [Figure 16] FIG. 10 is a schematic cross-sectional view showing a third step of the collection and placement method according to the present embodiment. [Figure 17] FIG. 10 is a schematic cross-sectional view showing a fourth step of the collection and placement method according to the present embodiment. [Figure 18]FIG. 10 is a schematic cross-sectional view showing a fifth step of the collection and placement method according to the present embodiment. [Figure 19] FIG. 19 is a schematic enlarged perspective view showing the plate included in FIG. 18 and the gel plate thereon. [Figure 20] FIG. 10 is a schematic cross-sectional view showing a sixth step of the collection and placement method according to the present embodiment. [Figure 21] FIG. 21 is a schematic enlarged cross-sectional view of the portion where the collection pin cuts off the gel plate immediately after FIG. 20. [Figure 22] FIG. 10 is a schematic enlarged cross-sectional view showing the seventh step of the collection and placement method according to the present embodiment. [Figure 23] FIG. 10 is a schematic enlarged cross-sectional view showing the eighth step of the collection and placement method according to the present embodiment. [Figure 24] FIG. 10 is a schematic cross-sectional view showing an example of a process in which the gel is dissolved from the gel sheet portion and only the cells are collected. [Figure 25] FIG. 23 is a schematic enlarged cross-sectional view in which the sample in FIG. 22 is replaced with a factor. [Figure 26] FIG. 26 is a schematic enlarged cross-sectional view showing the process of the gradual release or elution of a factor contained in a gel from the gel sheet portion of FIG. 25. [Figure 27] FIG. 10 is a schematic enlarged cross-sectional view showing a first step of supplying a high-viscosity solution into a container in a modified example. [Figure 28] FIG. 10 is a schematic enlarged cross-sectional view showing a second step of supplying a high-viscosity solution into a container in the modified example. [Figure 29] FIG. 10 is a schematic enlarged cross-sectional view showing a third step of supplying a high-viscosity solution into a container in the modified example. [Figure 30] FIG. 22 is a schematic enlarged cross-sectional view corresponding to FIG. 21 in a modified example in which a high-viscosity solution is supplied. [Figure 31] FIG. 23 is a schematic enlarged cross-sectional view corresponding to FIG. 22 in a modified example in which a high-viscosity solution is supplied. [Figure 32] FIG. 24 is a schematic enlarged cross-sectional view corresponding to FIG. 23 in a modified example in which a high-viscosity solution is supplied. [Figure 33] 10 is a photograph of the gel sheet portion collected with the first collection pin. [Figure 34]10 is a photograph of the gel sheet portion collected with the second collection pin. [Figure 35] 1 is a photograph of a gel sheet portion taken from the first gel sheet. [Figure 36] 10 is a photograph of a gel sheet portion taken from the second gel sheet. DETAILED DESCRIPTION OF THE INVENTION

[0016] Hereinafter, the present embodiment will be described with reference to the drawings.

[0017] (Collection placement components) First, the features of this embodiment will be briefly described. Referring to Figure 3, the collection and placement member (collection and placement mechanism 104A) of this embodiment comprises a collection pin 141, a placement pin 142, and a fixing holder 143. The collection pin 141 is hollow and can cut out and collect the material to be treated. The placement pin 142 can be inserted into the collection pin 141, and by pressing the collection pin 141, the cut out material to be treated can be pushed out of the collection pin 141 and placed in a desired position. The collection pin 141 is fixed to the fixing holder 143.

[0018] FIG. 1 is a schematic front view of the interior of a sampling and placement device according to this embodiment. For ease of explanation, the X, Y, and Z directions are introduced. Referring to FIG. 1, the sampling and placement device 100 is a device for sampling a sample from a gel plate 6, which serves as a specimen sample, as described below. The sampling and placement device 100 mainly comprises a processing chamber, a sample XY stage 101 arranged inside the processing chamber, a container XY stage 102, and a sampling and placement member 104. Although the sampling and placement device 100 in FIG. 1 has only one sampling and placement member 104, it may include multiple members.

[0019] The sample XY stage 101 is movable horizontally, i.e., along the X and Y directions (X and Y directions). Specifically, for example, a guide unit is provided on the underside of the sample XY stage 101. The guide unit is slidably connected to a guide rail provided on the bottom surface of the processing chamber. The upper surface of the sample XY stage 101 (holding table) serves as a mounting surface on which a gel plate 6 can be fixed. A stage penetration portion 101A that penetrates at least a portion of the sample XY stage 101 is formed. The gel plate 6 is preferably fixed at a position that overlaps the stage penetration portion 101A.

[0020] An XY stage 102 for containers is disposed below the XY stage 101 for samples. The XY stage 102 for containers is movable horizontally, i.e., along the X and Y directions (X and Y directions). Specifically, for example, a guide unit is disposed on the underside of the XY stage 102 for containers. The guide unit is slidably connected to a guide rail disposed on the bottom surface of the processing chamber. The XY stage 102 for containers fixes a plate 8 on which a container 9A capable of accommodating a portion of a gel plate 6 is formed. As a result, in FIG. 1, the collection and placement member 104, the gel plate 6, and the plate 8 on which the container 9A is formed are disposed in this order from top to bottom in the Z direction.

[0021] The sampling and positioning member 104 and the observation optical system 106 are connected to a member movable in the Z direction, such as a Z-axis table. In other words, the sampling and positioning member 104 and the observation optical system 106 are held within the sample sampling device 100 so that they can move in the Z direction. The observation optical system 106 observes and measures the position of the sample to be sampled contained in the gel plate 6. The observation optical system 106 may be equipped with a CCD camera that converts the observed image into an electrical signal. Observation of the gel plate 6 and the like using the observation optical system 106 may be performed using visible light. However, observation of the gel plate 6 and the like is not limited to visible light, and may also be performed using infrared light, X-rays, ultrasound, and the like. Depending on the material of the gel plate 6, magnetic observation of the gel plate 6 is also possible. A gel plate 6 observed by means other than visible light does not need to be transparent or translucent and may be opaque.

[0022] Although not shown, a control unit is installed outside the processing chamber. The control unit has a monitor, a control computer, and an operation panel. In the control unit, a command value for the needle movement speed for sampling from the operation panel is input and stored in the control computer's storage device. For example, during a sample sampling operation, the speed command value is read from the storage device and sent to the control program for the sampling placement member 104. The control program for the sampling placement member 104 determines the rotation speed of the motor included in the sampling placement member 104 based on the command value and moves the sampling pin 141 and the placement pin 142 up and down at a predetermined speed. This causes the sampling operation of the sampling placement member 104 to be performed. If the control computer communicates with a control system located further upstream, the command value may be received from that control system. In addition, parameters corresponding to the material of the gel sheet from which the sample is to be sampled may be stored in the storage device, and a command value for the sampling operation speed may be calculated depending on the specified gel sheet material, thickness, and sampling method.

[0023] (Configuration of collection and placement mechanism) FIG. 2 is a front view of the sampling and positioning mechanism constituting the sampling and positioning member of FIG. 1, as viewed from the negative side in the Y direction. FIG. 3 is a side view of the sampling and positioning mechanism of FIG. 2, as viewed from the negative side in the X direction. Referring to FIGS. 2 and 3, the sampling and positioning member 104 of FIG. 1 includes a part thereof, the sampling and positioning mechanism 104A. The sampling and positioning mechanism 104A mainly includes a sampling pin 141, a positioning pin 142, and a fixing holder 143. The sampling pin 141 is a pin (needle-shaped member) for cutting and sampling a material to be processed, including a sample, etc. The positioning pin 142 is a pin for releasing a sample, etc., sampled and held by the sampling pin 141, and placing it at a desired position. The sampling pin 141 and the positioning pin 142 are formed of, for example, a metal such as stainless steel, or plastic.

[0024] The sampling pin 141 has a sampling pin body 141A and a sampling pin holder 141B. The sampling pin body 141A is pin-shaped (pipe-shaped) and cylindrical with a hollow center in a cross section intersecting the extension direction (the vertical direction in Figures 2 and 3: Z direction). The sampling pin holder 141B is inserted onto the outer peripheral surface of the upper part of the sampling pin body 141A. Therefore, the sampling pin holder 141B also has a hollow center in a cross section intersecting the Z direction in Figures 2 and 3. The sampling pin holder 141B is fixed onto the outer peripheral surface of the sampling pin body 141A by press-fitting. Alternatively, the sampling pin holder 141B is adhered to the outer peripheral surface of the sampling pin body 141A with adhesive 141C. 2 and 3, the sampling pin holder 141B is arranged so that the top surface is along the XY plane, but the bottom surface may be inclined with respect to the XY plane, with the outer periphery having a smaller dimension in the Z direction than the inner periphery. As described above, the sampling pin 141 has a hollow shape as a whole.

[0025] The positioning pin 142 has a positioning pin main body 142A and a positioning pin holder 142B. The positioning pin main body 142A is pin-shaped and extends in the Z direction. The positioning pin main body 142A may, for example, be solid inside. However, it is sufficient that at least the bottom surface of the positioning pin main body 142A in the Z direction is closed so as not to have a hole in the center or the like, and the inside may be hollow. The positioning pin holder 142B has a larger dimension along the X direction (and Y direction) than the positioning pin main body 142A. The positioning pin holder 142B holds the positioning pin main body 142A. For example, the bottom surface of the positioning pin holder 142B may be connected to the top surface of the positioning pin main body 142A, and the two may be integrated.

[0026] The location pin body 142A has a smaller cross-sectional dimension than the hollow portion of the sampling pin body 141A. Therefore, the location pin body 142A can be inserted into the interior of the sampling pin body 141A. On the other hand, the location pin holder 142B has a larger cross-sectional dimension than the hollow portion of the sampling pin body 141A. Therefore, the location pin holder 142B cannot be inserted into the interior of the sampling pin body 141A.

[0027] The sampling pin body 141A presses the material to be processed by descending, cutting out a portion of the material, and can store the cut-out portion inside the hollow cylindrical portion. The material to be processed is a gel sheet 4, which is a sheet of gel 4D that supports the sample 4B, the object to be sampled (see FIG. 16, described below). The positioning pin body 142A then moves inside the sampling pin body 141A. The positioning pin body 142A contacts the material to be processed inside the sampling pin 141 and presses against it. This causes the positioning pin body 142A to push the material to the outside of the sampling pin 141. The positioning pin body 142A can move up and down inside the hollow portion of the sampling pin body 141A. In other words, the positioning pin body 142A passes through the interior of the sampling pin body 141A. The positioning pin body 142A moves up and down by a first drive unit 144, described below. This allows the cut material to be placed in a desired position.

[0028] A spring 142C is installed in the area of ​​the interior of the positioning pin holder 142B adjacent to the positioning pin main body 142A, i.e., the lowest area within the positioning pin holder 142B. The spring 142C connects the positioning pin holder 142B to the top of the positioning pin main body 142A. The spring 142C absorbs the Z-direction force applied to the positioning pin holder 142B when the positioning pin main body 142A collides with the material to be processed, such as a substrate. The positioning pin 142 does not need to have the spring 142C. If the positioning pin 142 does not have the spring 142C, it will not be able to absorb the force during a collision. However, if the positioning pin 142 does not have the spring 142C, the effects of a collision can be eliminated by precisely controlling the descending position of the positioning pin 142.

[0029] The fixing holder 143 can hold the sampling pin 141. The fixing holder 143 is disposed above the sampling pin 141 in FIG. 2 and on the rear side of the positioning pin 142. The fixing holder 143 has, for example, a plate shape (rectangular parallelepiped shape), but is not limited to this. The sampling pin 141 is fixed to the fixing holder 143. Specifically, the top surface of the sampling pin holder 141B and the bottom surface of the fixing holder 143 are in contact with and fixed to each other at the fixing portion FXD. The fixing portion FXD may be detachably fixed by, for example, a stainless steel screw screwed near the fixing portion FXD of the sampling pin holder 141B and the magnetic force of a magnet embedded near the fixing portion FXD of the fixing holder 143. Alternatively, the fixing portion FXD may be detachably fixed by screwing the stainless steel screw screwed into the sampling pin holder 141 to the fixing holder 143.

[0030] A first driving unit 144 is provided on a front surface 143f (the surface on the negative side in the Y direction) of the fixing holder 143, above the positioning pin holder 142B in the Z direction. The first driving unit 144 includes a first motor 144A, a disk member 144B, and a link member 144C. The first driving unit 144 is attached to a positioning pin holder fixing portion 145.

[0031] The positioning pin holder fixing portion 145 has a first portion 145a and a second portion 145b. The first portion 145a has a plate shape and is installed so that its main surface on the back side contacts the front surface 143f of the fixing holder 143 and is slidable on the front surface 143f. The second portion 145b is a plate-shaped portion installed on the main surface on the front side opposite the main surface of the first portion 145a that contacts the front surface 143f, so as to be approximately perpendicular to the first portion 145a. The front surface 143f of the fixing holder 143 may be formed with, for example, a groove extending along the Z direction, along which the positioning pin holder fixing portion 145 slides.

[0032] The link member 144C is installed on the front side of the disc member 144B in Figure 2. The link member 144C has an elongated shape extending in the vertical direction, i.e., the Z direction. The main surface on the back side of the disc shape of the disc member 144B is in contact with the front surface 143f of the fixing holder 143. A first motor 144A is attached at a position overlapping with the circular center of the disc member 144B. At least a portion of the first motor 144A may be disposed within the fixing holder 143. The disc member 144B can be rotated around its center by the first motor 144A. A first end, which is one end of the link member 144C in the extension direction, is fixed to a part of the outer periphery of the disc member 144B.

[0033] A second end (the lower end in FIG. 2) of the link member 144C opposite the first end in the extension direction (Z direction) is fixed to the positioning pin holder fixing portion 145 so as to come into contact with the first portion 145a of the positioning pin holder fixing portion 145. The second end may be capable of coming into contact with the second portion 145b of the positioning pin holder fixing portion 145. As the first motor 144A rotates the disc member 144B, the link member 144C moves up and down, and the second end moves the positioning pin 142 up and down. This will be described in detail later.

[0034] To achieve this operation, the positioning pin holder fixing portion 145 is provided between the second end and the positioning pin 142 (positioning pin holder 142B), and the positioning pin 142 is fixed thereto. The positioning pin 142 is fixed so as to come into contact with both the first portion 145a and the second portion 145b of the positioning pin holder fixing portion 145. The positioning pin 142 (positioning pin holder 142B) and the positioning pin holder fixing portion 145 (first portion 145a and second portion 145b) may be detachably fixed to each other by, for example, magnetic force between a magnet embedded in the positioning pin holder 142B and a magnet embedded in the first portion 145a. Alternatively, the two may be detachably fixed to each other by screw fastening.

[0035] (Overall configuration of collection placement components) 4 is a front view of the entire collection and placement member of FIG. 1, as viewed from the negative side in the Y direction. FIG. 5 is a side view of the collection and placement member of FIG. 4, as viewed from the negative side in the X direction. Referring to FIGS. 4 and 5, the collection and placement member 104 further includes a second drive unit 146 in addition to the collection and placement mechanism 104A of FIGS. 2 and 3. The second drive unit 146 is disposed on the rear side of the fixed holder 143 of FIGS. 2 and 3, and includes a movable holder 146A. The movable holder 146A has, for example, a plate shape (rectangular parallelepiped shape) and is configured to store components therein, but is not limited to this. The movable holder 146A has a larger dimension in the Z direction than the fixed holder 143. The collection and placement mechanism 104A is attached to the movable holder 146A.

[0036] The second driving unit 146 is a slide mechanism that slides the collection and placement mechanism 104A, including the fixing holder 143, in the Z direction. In addition to the movable holder 146A, the second driving unit 146 also includes a second motor 146B, a vertically extending screw 146C, and a screw connecting unit 146D. The vertically extending screw 146C extends in the up-down direction and has a male thread formed on its outer periphery. The vertically extending screw 146C is disposed within the rectangular parallelepiped movable holder 146A, for example, and has a larger dimension in the Z direction than the fixing holder 143. The vertically extending screw 146C may be housed in a groove that is formed in the front surface 146f of the movable holder 146A and extends in the Z direction. The second motor 146B is attached to one end (upper end) of the vertically extending screw 146C within the movable holder 146A. The screw connecting portion 146D is attached to the back surface 143b (the surface opposite to the front surface 143f) of the fixing holder 143 so as to be able to connect the vertically extending screw 146C and the fixing holder 143. The screw connecting portion 146D has a female thread formed thereon, and this female thread is fastened to the male thread of the vertically extending screw 146C.

[0037] The vertically extending screw 146C is rotated by the second motor 146B. Although the vertically extending screw 146C does not move vertically when rotated, the screw connector 146D fastened to the vertically extending screw 146C moves vertically. As a result, the entire collection and placement mechanism 104A, including the fixing holder 143 and collection pin 141 attached to the screw connector 146D, moves vertically. This will be described in detail later.

[0038] (Collection process) FIG. 6 is a front view of the collection and placement member of FIG. 4, showing the operation of the process of cutting and collecting the material to be processed, as viewed from the negative side in the Y direction. FIG. 7 is a side view of the operation of FIG. 6, as viewed from the negative side in the X direction. Referring to FIGS. 6 and 7, the rotational motion of the second motor 146B is converted into vertical motion of the screw connection part 146D. The screw connection part 146D and the rear surface 143b of the fixing holder 143 are fixed by screws. Therefore, vertical movement of the screw connection part 146D moves the entire collection and placement mechanism 104A, including the fixing holder 143 and the collection pin 141, in the vertical direction. Note that a slide groove 146E (see FIG. 6) extending along the Z direction is preferably formed on the front surface 146f of the movable holder 146A, which faces the rear surface 143b of the fixing holder 143, to allow the screw connection part 146D to slide. The slide groove 146E is formed so that its depth extends from the front surface 146f toward the positive side in the Y direction (digging from the front side to the back side of the paper in FIG. 6). In this case, the screw connecting portion 146D is arranged so that at least a portion thereof is fitted into the slide groove 146E.

[0039] For example, the sampling and positioning mechanism 104A moves downward as indicated by arrow M1 in Figures 6 and 7. As a result, the tip of the sampling pin 141 (sampling pin body 141A) cuts out the material to be processed that is positioned below the tip. The cut-out portion of the material to be processed is stored in the hollow portion of the sampling pin body 141A.

[0040] (Placement process) FIG. 8 is a front view of the sampling and placement mechanism of FIG. 2, showing the operation of the first step of releasing and placing the material to be processed, as viewed from the negative side in the Y direction. Referring to FIG. 8, a first end of a link member 144C is fixed to a portion of the outer periphery of a disc member 144B by a joint 144D1. A second end of the link member 144C is fixed to the positioning pin holder fixing portion 145 by a joint 144D2. In the initial state of FIG. 8, the first end of the link member 144C is positioned at the top of the disc member 144B. Therefore, the positioning pin holder fixing portion 145 and the positioning pin 142 connected to the link member 144C are positioned at the top in the Z direction. The tip (bottom) of the positioning pin main body 142A is positioned above the bottom of the sampling pin main body 141A. In this state, the bottom of the sampling pin main body 141A becomes the bottom of the entire assembly, and this portion can be used to cut out the material to be processed.

[0041] 9 is a front view of the operation in the second step following FIG. 8, seen from the negative side in the Y direction. Referring to FIG. 9, the first motor 144A (see FIG. 3) rotates the disk member 144B in the direction indicated by the arrow R in the figure. As a result, the first end of the link member 144C is positioned at the center position of the disk member 144B in the Z direction. Therefore, the link member 144C is tilted slightly with respect to the Z direction, and the positioning pin holder fixing portion 145 and the positioning pin 142 connected thereto are lowered as indicated by the arrow M2 in the figure.

[0042] FIG. 10 is a front view of the operation in the third step following FIG. 9, viewed from the negative Y-axis direction. FIG. 11 is a side view of the operation in FIG. 10, viewed from the negative X-axis direction. Referring to FIGS. 10 and 11, the disk member 144B further rotates from the state shown in FIG. 9, and the first end reaches the bottom of the disk member 144B. At this time, as indicated by arrow M2 in the figure, the positioning pin 142 moves to the lowest position relative to the sampling pin 141. As the positioning pin 142 moves downward, the tip of the positioning pin body 142A slides downward to penetrate the hollow portion of the sampling pin body 141A. As a result, the tip of the positioning pin body 142A contacts and presses downward against the material to be treated housed within the sampling pin body 141A, ejecting it from the sampling pin body 141A and positioning it in the desired position.

[0043] Fig. 12 is a front view, seen from the negative side in the Y direction, of the operation in a fourth step following Fig. 10. Fig. 13 is a front view, seen from the negative side in the Y direction, of the operation in a fifth step following Fig. 12. Referring to Figs. 12 and 13, when the disk member 144B rotates further, the movement direction of the positioning pin 142 and the positioning pin holder fixing portion 145 is reversed as shown by arrow M3 in the figures, and they return to the initial state of Fig. 8 as shown in Fig. 13.

[0044] As a modified example instead of joints 144D1 and 144D2, the following configuration may be used: For example, a spring may be provided to connect positioning pin holder fixing portion 145 and link member 144C, and when link member 144C presses downward, positioning pin holder fixing portion 145 descends, but when link member 144C is not pressed, positioning pin holder fixing portion 145 rises due to the biasing force of the spring.

[0045] (Details of how to collect and place the object) FIG. 14 is a schematic cross-sectional view showing a first step of the collection and placement method according to this embodiment. Referring to FIG. 14, a film 2 is placed on, for example, a glass slide 1. For example, the glass slide 1 is a thin glass plate having a thickness of approximately 1.2 mm. The film 2 is made of a soft material and is thin so that it can be easily broken by puncturing with a needle. For example, the film 2 is made of polyvinylidene chloride having a thickness of approximately 10 μm. For example, a thin film used for food packaging or a thin film of silicone rubber is preferably used as the film 2. A spacer 3 is placed on the film 2. The spacer 3 may be made of the same glass material as the glass slide 1. The spacer 3 may have a thickness of 100 μm or more and 200 μm or less. For example, the thickness of the spacer 3 may be 100 μm or 200 μm. A through-hole 3A is formed in the center of the spacer 3 when viewed from above (in a plan view) in FIG. 14. The through-hole 3A penetrates the spacer 3. Therefore, the inside of through-hole 3A is hollow and is not filled with glass or other materials that constitute spacer 3. Through-hole 3A may have any planar shape, such as a rectangular shape.

[0046] Next, a fluid sample 4C, which includes one or more samples 4B as the target object and a fluid gel raw material 4A capable of forming a gel, is supplied into the through-hole 3A. The sample 4B is, for example, a cell to be collected. In FIG. 14, multiple samples 4B are arranged one by one at intervals. The gel raw material 4A is, for example, a 1.5% agarose gel in which a fluorescent dye is added to 1.5 mL of a gel precursor. However, the gel raw material 4A is not limited to this, and may also be, for example, collagen or a mixture of alginate and collagen.

[0047] 15 is a schematic cross-sectional view showing a second step of the collection and placement method according to the present embodiment. Referring to FIG. 15, a film 2 is attached to the underside of a glass slide 5. The glass slide 5 may be made of the same material and have the same size as the glass slide 1 in FIG. 14. The film 2 on the upper side of the flowing sample 4C in FIG. 15 may be made of the same material and have the same size as the film 2 in FIG. 14.

[0048] FIG. 16 is a schematic cross-sectional view showing the third step of the collection and placement method according to this embodiment. Referring to FIG. 16, a glass slide 5 with a film 2 attached thereto is placed on a spacer 3. The film 2 attached to the glass slide 5 comes into contact with the flowing sample 4C and the spacer 3. The glass slide 5 and film 2 press the flowing sample 4C and the spacer 3 downward. As a result, the flowing sample 4C and the spacer 3 are sandwiched between the glass slide 1 with a film 2 attached thereto below and the glass slide 5 with a film 2 attached thereto above. As a result, the glass slide 1, film 2, spacer 3 and flowing sample 4C, film 2, and glass slide 5 are stacked in this order from bottom to top.

[0049] Next, for example, if the gel raw material 4A is a material that solidifies depending on temperature, the entire layered structure is cooled to 4°C. By cooling, the gel raw material 4A that constitutes the fluid sample 4C solidifies into a gel 4D, and a gel sheet 4 is formed in the through-hole 3A with the sample 4B supported inside the gel 4D. Although the gel 4D is solidified, it is soft enough to be easily broken by puncturing with a needle. Note that "the sample 4B is supported by the gel 4D" means, for example, that the sample 4B is embedded so that it is surrounded by and buried in the hard gel 4D, and the sample 4B is in a non-fluid state.

[0050] FIG. 17 is a schematic cross-sectional view showing a fourth step of the collection and placement method according to this embodiment. Referring to FIG. 17, after the gel sheet 4 is provided, the film 2 and slide glass 5 placed on top of the gel sheet 4 are removed, and the slide glass 1 and spacer 3 are also removed. This results in a gel plate 6 including the film 2 and the gel sheet 4 placed on the film 2. The gel sheet 4 is formed by supporting the sample 4B to be collected in a solidified gel 4D, for example, by embedding it therein. The gel 4D is formed in a sheet-like form on the film 2. In other words, the gel sheet 4 is formed in a sheet-like form on the film 2 and includes the gel 4D that supports the sample 4B to be collected. In this embodiment, "providing the gel sheet 4" refers to the process of obtaining the gel sheet 4 by supporting the sample 4B in the gel 4D, for example, by embedding it therein, being performed on the film 2. Alternatively, a gel sheet 4 previously formed externally may be attached to the film 2. The gel sheet 4 can be formed in advance on any flat surface.

[0051] Figure 18 is a schematic cross-sectional view showing the fifth step of the collection and placement method according to this embodiment. Figure 19 is a schematic enlarged perspective view showing the plate included in Figure 18 and the gel plate thereon. With reference to Figures 18 and 19, the formed gel plate 6 is punctured by a collection pin 141 descending from above. The mechanism by which the collection pin 141 descends during puncturing is as described above (Figures 6 to 7).

[0052] Meanwhile, the gel plate 6 is placed on the plate 8. The plate 8 is a cell culture plate. A plurality of containers 9A are formed on the plate 8 as collection sections. The plurality of containers 9A are recessed portions of the upper surface of the plate 8. Because of their recessed shape, a portion of the cut gel sheet 4 (gel sheet portion 4E, described below) can be stored in the container 9A. After being cut, the portion of the cut gel sheet 4 (gel sheet portion 4E, described below) may be pulled out from the container 9A and the film 2 while still held in the needle (collection pin body 141A) and stored in another prepared container or the like before being placed.

[0053] The multiple containers 9A may be spaced apart from one another, for example, in 8 rows in the depth direction and 12 rows in the width direction in FIG. 19, for a total of 96 containers. The containers 9A may have any planar shape, such as a circle. In FIGS. 18 and 19, the film 2 constituting the gel plate 6 is in contact with the upper surface of the plate 8 so as to cover at least a portion of the multiple containers 9A. To achieve this, a portion (upper region) of the plate 8 may be disposed within the stage penetration portion 101A in FIG. 1.

[0054] For example, a sheet fixing member 21 may be placed above the gel plate 6. The sheet fixing member 21 prevents the gel plate 6 from shifting out of position due to force being applied from above to the gel 4D during the process of cutting out the gel sheet 4. Instead of the sheet fixing member 21, an adhesive film tape may be used as a means for preventing the gel plate 6 from shifting out of position.

[0055] Plate 8, on which vessel 9A as a culture vessel is formed, is made of any material selected from the group consisting of polystyrene, polypropylene, polycarbonate, and polyethylene terephthalate.

[0056] FIG. 20 is a schematic cross-sectional view showing the sixth step of the sampling and positioning method according to this embodiment. FIG. 21 is a schematic enlarged cross-sectional view of the portion where the sampling pin cuts off the gel plate immediately after the step shown in FIG. 20. Referring to FIGS. 20 and 21, the entire sampling and positioning member 104, including the sampling pin 141, is lowered from the state shown in FIG. 18. The position to which the sampling pin 141 should be lowered is the position of the target sample 4B to be sampled, as determined from an image acquired using an observation optical system (such as the observation optical system 106 in FIG. 1). For example, as shown in FIG. 8, the sampling pin body 142A is positioned at the top, and the bottom of the sampling pin body 141A is positioned below the bottom of the sampling pin body 142A. The sampling pin body 141A punctures the gel plate 6, particularly the gel sheet 4. At this time, it is preferable that the volume of at least the hollow portion of the sampling pin body 141A (the hollow portion from the bottom of the sampling pin body 141A to the bottom of the sampling pin body 142A) be large enough to accommodate the cut gel sheet portion.

[0057] The punctured portion of the gel sheet 4 is damaged by the passage of the sharp tip of the sampling pin body 141A. The sampling pin body 141A penetrates the gel sheet 4, tearing it apart. The portion of the gel sheet 4 cut away by the sampling pin body 141A from the original gel sheet 4 is the gel sheet portion 4E. The cut gel sheet portion 4E preferably comprises a gel 4D and a sample 4B embedded therein. This prevents the sample 4B from flowing relative to the gel 4D. This prevents the sample 4B from collapsing and scattering when the gel sheet portion 4E is collected and placed in the sampling pin 141. The cut gel sheet portion 4E is collected (stored) within the sampling pin body 141A.

[0058] After the gel sheet portion 4E is cut off by the sampling pin body 141A, the sampling pin body 141A further descends while holding the gel sheet portion 4E inside (hollow portion). As a result, the sampling pin body 141A penetrates the portion of the film 2 located directly below the gel sheet 4 on the gel plate 6. As a result, the film 2 is also damaged in the same way as the gel sheet 4.

[0059] FIG. 22 is a schematic enlarged cross-sectional view showing the seventh step of the sampling and positioning method according to this embodiment. Referring to FIG. 22, after FIG. 21, the positioning pin 142 is lowered in the direction indicated by the arrow in the figure. The positioning pin 142, particularly the positioning pin body 142A, is inserted through the hollow portion inside the sampling pin body 141A. FIG. 22 corresponds to the state shown in FIGS. 10 and 11 above, for example. The lowered positioning pin body 142A contacts and presses downward against the gel sheet portion 4E collected by the sampling pin body 141A. This pushes the gel sheet portion 4E out of the sampling pin body 141A and places it outside the sampling pin 141A. Specifically, the gel sheet portion 4E is collected on the inside (bottom) of the container 9A. The bottom of the container 9A may be a dry surface that is not coated with a highly viscous solution or the like.

[0060] Because the sampling pin body 141A penetrates the gel plate 6, the sampling pin body 141A breaks through both the gel sheet 4 and the film 2. However, as shown in Figure 21, it is preferable that only the gel sheet 4 is stored within the hollow shape portion 7C, and that the film 2 is not stored within the hollow shape portion 7C. As a result, as shown in Figure 22, it is preferable that only the gel sheet portion 4E is collected within the container 9A, and that the film 2 is not collected within the container 9A.

[0061] 23 is a schematic enlarged cross-sectional view showing the eighth step of the collection and placement method according to the present embodiment. Referring to FIG. 23, after the gel sheet portion 4E is placed on the bottom surface of the container 9A, the placement pin body 142A rises again. FIG. 23 corresponds to the state shown in FIGS. 12 and 13, for example.

[0062] (material) Gel 4D is a gel formed from biocompatible gel raw material 4A. Here, "biocompatible" means that it does not cause inflammation or other toxicity when contacted with living tissue such as cells, and has low invasiveness to cells. This can suppress damage to cells (specimen 4B) caused by the gel. Gel raw material 4A is preferably transparent or translucent, so that the position of embedded specimen 4B can be observed. From the viewpoint of satisfying the above, gel raw material 4A is any material selected from the group consisting of collagen, gelatin, fibrin, sodium alginate, gellan gum, agarose, hyaluronic acid, chitin, chitosan, polyethylene glycol, polyvinyl alcohol, and 2-methacryloyloxyethyl phosphorylcholine (MPC polymer).

[0063] The sample 4B as the object may be, for example, an isolated cell or a cell aggregate. Here, "isolated" refers to both the case where only one sample 4B is contained within the gel sheet portion 4E as shown in Figures 21 to 23, and the case where multiple samples 4B are contained, one by one, spaced apart from each other, as shown in the left half of the gel raw material 4A in Figures 14 and 15. On the other hand, a mass of multiple samples 4B, as shown in the right half of the gel raw material 4A in Figures 14 and 15, is called an aggregate. The cells as sample 4B may be any one selected from the group consisting of primary cultured cells, established cell lines, and stem cell-derived differentiated cells, including cardiomyocytes, stem cells, neurons, and fibroblasts.

[0064] For example, if multiple samples 4B gather in the gel sheet 4 to form an aggregate, particularly if the aggregate is large, there is a risk that the sampling pin body 141A may puncture the sample 4B and damage the sample 4B. Damage to the sample 4B may make it difficult to analyze, for example, the cross section of the tissue. Therefore, as described above, it is preferable to disperse the multiple samples 4B so that they are spaced apart from each other. This reduces damage to the samples 4B due to puncturing. However, if the aggregate is particularly small in size, can pass through the hollow portion of the sampling pin body 141A, and there is little chance that the sampling pin body 141A will puncture the sample 4B, then the formation of an aggregate is acceptable.

[0065] 24 is a schematic cross-sectional view showing an example of a process in which the gel is dissolved from the gel sheet portion to recover only the cells. Referring to FIG. 24, it is also possible to recover only the cells (sample 4B) by dissolving the gel 4D that supports and fixes the sample 4B from the cut gel sheet portion 4E. Specifically, for example, 5×10 5 A gel sheet portion 4E containing fibroblasts at 0.1 cells / mL is placed inside the gel 4D. A gel solubilizer 10 is supplied into the device. The gel solubilizer 10 dissolves the gel 4D. As a result, as shown in the lower diagram of Figure 24, the gel solubilizer 10 contains the dissolved gel 4D and accumulates inside the device as gel solubilizer 10A. Because this is liquid, it can be discarded to recover only the sample 4B. For example, if the gel 4D uses sodium alginate, an alginate-degrading enzyme or an EDTA (ethylenediaminetetraacetic acid) solution is used as the gel solubilizer 10.

[0066] FIG. 25 is a schematic enlarged cross-sectional view in which the sample in FIG. 22 is replaced with a factor. Referring to FIG. 25, gel sheet portion 4H is shown instead of gel sheet portion 4E, and factor 4G is shown instead of sample 4B. In the collection and placement methods of FIGS. 14 to 23, the object embedded in gel 4D may be any factor 4G selected from the group consisting of proteins, peptides, compounds, and particles. For this reason, the symbol 4G in parentheses is added to FIGS. 14 to 17. The factor 4G protein may be any one selected from the group consisting of fibroblast growth factor (FGF), hepatocyte growth factor (HGF), nerve growth factor (NGF), and vascular endothelial growth factor (VEGF) as growth factors. Alternatively, the factor 4G protein may be glucosidase alpha as an enzyme. Furthermore, the factor 4G protein may be a cytokine such as interleukin-2 (IL-2) or granulocyte colony-stimulating factor (G-CSF).The factor 4G peptide may be either a hormone or a neuropeptide.

[0067] The compound of factor 4G may be any one selected from the group consisting of adrenergic β1 receptor agonists / antagonists, Rho kinase inhibitors, and Wnt signaling pathway-related inhibitors. The particle of factor 4G may be made from any one selected from the group consisting of plastic, lipid bilayer membrane, and polysaccharide. These factor 4G proteins, peptides, and compounds thereof, as well as proteins, peptides, and compounds thereof adsorbed or encapsulated in particles, can be sustained-released as described below.

[0068] FIG. 26 is a schematic enlarged cross-sectional view showing the process of gradually releasing or eluting a factor encapsulated in a gel from the gel sheet portion of FIG. 25. Referring to FIG. 26, a gel solubilizer is supplied to the gel sheet portion 4H, which embeds factor 4G and is placed in the container 9A of FIG. 25, in the same manner as in FIG. 24. The gel solubilizer dissolves the gel 4D, and the gel 4D accumulates in the container 9A as gel solubilizer 10A. Within this gel solubilizer 10A, the factor 4G contained in the gel sheet portion 4H is gradually released from the gel sheet portion 4H, forming a concentration gradient 15 of the factor 4G. A difference in concentration of the factor 4G occurs between the gel sheet portion 4H embedding the factor 4G and the solution 10A, which does not contain the factor 4G. This concentration difference causes the factor 4G to disperse. The factor 4G contained in the gel sheet portion 4H is gradually released from the gel sheet portion 4H, forming a concentration gradient 15 of the factor 4G. That is, after the factor 4G is released from the sampling pin main body 141A as shown in Figure 25, the factor 4G contained in the gel sheet portion 4H can be gradually released as shown in Figure 26. In Figure 26, the factor 4G may be gradually released by dissolving the gel 4D as described above, but the factor 4G may also be eluted. Elution means that the factor 4G contained in the gel 4D dissolves without dissolving the gel 4D.

[0069] (Action and effect) The collection and placement member 104 according to the present disclosure comprises a collection pin 141, a placement pin 142, and a fixing holder 143. The collection pin 141 is hollow and can cut out and collect the material to be processed. The placement pin 142 can be inserted into the collection pin 141, and when pressed by the collection pin 141, it comes into contact with the material to be processed cut out inside the collection pin 141 and presses the material to be processed, thereby placing the material to be processed outside the collection pin 141. The collection pin 141 is fixed to the fixing holder 143. The material to be processed is a gel sheet 4 in the form of a sheet of gel 4D that supports a sample 4B, which is the object to be collected.

[0070] The material to be treated cut by the collection pin 141 is pushed out by the placement pin 142 and placed in the desired position (e.g., inside the container 9A). This prevents problems such as the collapse and scattering of objects such as the specimen 4B in the material to be treated, which are a concern when the material to be treated is discharged from the collection pin 141 using the pressure of a liquid, etc. This improves the stability of the collection process of the specimen 4B, etc., and improves the positional accuracy and reproducibility in the placement process of the specimen 4B, etc.

[0071] Furthermore, the sampling pin 141 is fixed to a fixing holder 143. This reduces the possibility of the sampling pin 141 moving unintentionally when an external vertical force is applied to the pin. This allows the target object to be cut out with high stability and reproducibility.

[0072] In the collection and placement member 104, the placement pin 142 can be inserted into the collection pin 141 while being moved vertically by the first drive unit 144. This allows the placement pin 142 to eject the gel sheet portion 4E stored in the collection pin 141 and place it in a desired position.

[0073] In the collection and placement member 104, the first drive unit 144 includes a first motor 144A, a rotatable disk member 144B attached to the first motor 144A, and a link member 144C extending vertically with a first end fixed to a portion of the outer periphery of the disk member 144B. A second end of the link member 144C, located opposite the first end in the extension direction, moves vertically as the first motor 144A rotates the disk member 144B, causing the second end to move the placement pin 142 vertically. The first drive unit 144 converts the rotation of the first motor 144A and the disk member 144B into a reciprocating motion in the vertical direction via the link member 144C. This allows the placement pin 142 to be inserted into the hollow portion of the collection pin 141, enabling the gel sheet portion 4E to be placed in the desired position.

[0074] The collection and positioning member 104 further includes a positioning pin holder fixing portion 145 fixed to the positioning pin 142 between the second end and the positioning pin 142. The positioning pin holder fixing portion 145 allows the positioning pin 142 to be stably fixed to the wiring pin holder fixing portion 145.

[0075] In the collection and positioning member 104, the positioning pin 142 includes a positioning pin main body 142A and a positioning pin holder 142B that holds the positioning pin main body 142A. A spring 142C is installed in an area inside the positioning pin holder 142B adjacent to the positioning pin main body 142A. This provides the effect of stably positioning the gel sheet portion 4E or the like in a desired position using the positioning pin main body 142A.

[0076] The collection and placement member 104 further includes a second drive unit 146 capable of moving the fixing holder 143 in the vertical direction. The second drive unit 146 includes a second motor 146B, a vertically extending screw 146C that extends in the vertical direction and has the second motor 146B attached to one end (e.g., the upper end) thereof, and a screw connector 146D that connects the vertically extending screw 146C to the fixing holder 143 and is slidable along the extension direction of the vertically extending screw 146C. The second drive unit 146 converts the rotational motion of the second motor 146B and the vertically extending screw 146C into a reciprocating motion in the vertical direction via the screw connector 146D. This allows the entire collection and placement mechanism 104A, including the collection pin 141, to reciprocate in the vertical direction, making it possible to cut out the desired portion of the gel sheet 4 with high positional accuracy.

[0077] The sampling and placement device 100 according to the present disclosure includes the sampling and placement member 104 described above, and a holding stage (XY stage for sample 101) that holds the material to be processed to be cut out by the sampling pin 141. The sampling and placement device 100 configured in this manner exhibits the effects provided by the sampling and placement member 104 described above.

[0078] In the collection and placement method according to the present disclosure, the collection and placement member 104 is prepared. By lowering the tip of the collection pin 141, the material to be treated that is placed below the collection pin 141 is cut and placed into the collection pin 141, and by lowering the tip of the placement pin 142, the material to be treated is brought into contact with and pressed against the material to be treated inside the collection pin 141, and the material to be treated is placed outside the collection pin 141.

[0079] The tip of the placement pin 142 comes into contact with and presses against the material to be treated, thereby releasing the material. This prevents problems such as the collapse and scattering of objects within the material to be treated, such as the specimen 4B, which can occur when the material to be treated is ejected from the collection pin 141 using liquid pressure or the like. This improves the stability of the collection process for the specimen 4B, etc., and improves the positional accuracy and reproducibility of the placement process for the specimen 4B, etc.

[0080] In the collection and placement method according to the present disclosure, a gel supporting the objects to be collected (sample 4B, factor 4G) is formed in a sheet shape on a film 2, and a gel sheet 4 is provided. The gel sheet 4 on the film 2 is punctured with a hollow collection pin 141, and gel sheet portions 4E and 4H, which are part of the gel sheet 4, are cut out, and the gel sheet portions 4E and 4H collected on the collection pin 141 are pushed out from inside the collection pin 141 by the pressure of the placement pin 142 and placed outside the collection pin 141. In the step of placing the gel outside the collection pin 141, the placement pin 142 is inserted through the interior of the collection pin 141.

[0081] When the placement pin 142 presses against the gel sheet portions 4E, 4H, the gel sheet portions 4E, 4H are pushed out from within the collection pin 141 and released to the outside. This prevents problems such as the collapse and scattering of objects such as the sample 4B, which are a concern when using liquid pressure to eject the gel sheet portions 4E, 4H from the collection pin 141. This improves the stability of the object collection process and improves the positional accuracy and reproducibility of the object placement process. The collection and placement device 100, to which this collection and placement method can be applied, may include a fixing holder 143 to which the collection pin 141 is fixed.

[0082] In the step of disposing the target outside the sampling pin 141, the sampling pin 141 cuts out the gel sheet portions 4E, 4H so as to include the target object, and then the sampling pin 141 penetrates the film 2 while holding the gel sheet portions 4E, 4H inside. According to the above method, only the desired gel sheet portions 4E, 4H are held inside the sampling pin 141 so as not to mix with the film 2, and this can be disposed in a desired position such as in the container 9A.

[0083] (Variation) FIG. 27 is a schematic enlarged cross-sectional view showing a first step of supplying a high-viscosity solution into a container in a modified example. FIG. 28 is a schematic enlarged cross-sectional view showing a second step of supplying a high-viscosity solution into a container in a modified example. FIG. 29 is a schematic enlarged cross-sectional view showing a third step of supplying a high-viscosity solution into a container in a modified example. With reference to FIGS. 27 to 29, a coating needle 7I of a commonly known coating mechanism is used to supply a high-viscosity solution 12 into a container 9A (for example, to the bottom surface). The coating needle 7I may be solid or hollow. Alternatively, a droplet dropping mechanism using a commonly known dispenser may be used to drop the high-viscosity solution 12 onto the bottom surface of the container 9A from above in the Z direction, as in FIGS. 27 to 29.

[0084] As the high-viscosity solution 12, it is preferable to use, for example, cellulose, cellulose nanofiber, methyl cellulose, carboxymethyl cellulose, hydroxybutyl cellulose, sodium alginate, sodium hyaluronate, polyethylene glycol, or the like.

[0085] FIG. 30 is a schematic enlarged cross-sectional view corresponding to FIG. 21 of a modified example in which a high-viscosity solution is supplied. FIG. 31 is a schematic enlarged cross-sectional view corresponding to FIG. 22 of a modified example in which a high-viscosity solution is supplied. FIG. 32 is a schematic enlarged cross-sectional view corresponding to FIG. 23 of a modified example in which a high-viscosity solution is supplied. With reference to FIGS. 30 to 32, the gel sheet portion 4E is placed on the high-viscosity solution 12 supplied into the container 9A by the collection and placement member 104, as in FIGS. 21 to 23. This allows the gel sheet portion 4E to be fixed more firmly than when it is placed on the dry surface of the container 9A as in FIGS. 21 to 23.

[0086] Alternatively, although not shown, the gel sheet portion 4E collected by the collection pin 141 can be supplied into the liquid filled in the container 9A. In this case, at least a portion of the tip of the collection pin 141 containing the gel sheet portion 4E is immersed in the liquid and adjusted to the desired position for discharging the gel sheet portion 4E. In this state, the gel sheet portion 4E is released into the liquid by pressing the positioning pin 142. By adjusting the viscosity of the liquid, the gel sheet portion 4E can be fixed within the container 9A, for example.

[0087] Furthermore, the sample 4B in this embodiment may be the above-mentioned cells. However, instead of cells, the sample 4B may be a biocompatible material in a granular or other form. The biocompatible material here is, for example, the aforementioned gel raw material 4A. The gel raw material 4A may be a biological material. In this case, the gel raw material 4A is granulated and suspended within the sheet-like gel 4D. Furthermore, the sheet-like gel 4D cut from the gel sheet 4 of this embodiment can also be applied to an evaluation chip for drug response evaluation. More specifically, the sheet-like gel 4D cut from the gel sheet 4 can be applied to an evaluation chip, particularly for drug discovery and development. [Example]

[0088] The material to be processed was collected and placed using the collection and placement device 100 shown in the above embodiment. The material to be processed was a gel sheet 4, which was a sheet of 5% agarose gel without a sample embedded inside, and was placed on a film 2 as shown in Figure 17 to form a gel plate 6. The film 2 was made of polyvinylidene chloride with a thickness of approximately 10 μm.

[0089] Next, as shown in Figure 21, the sampling pin 141 was lowered almost vertically from above, penetrating the gel sheet 4 and film 2 and collecting the gel sheet portion 4E. Two types of sampling and placement members 104 were used: one with a sampling pin body 141A (hereinafter referred to as the first sampling pin) having an inner diameter of 0.7 mm and an outer diameter of 1 mm, and the other with a sampling pin body 141A (hereinafter referred to as the second sampling pin) having an inner diameter of 1.1 mm and an outer diameter of 1.5 mm. The inner diameter and outer diameter here refer to diameters.

[0090] 22, a positioning pin body 142A with an outer diameter of 0.5 mm was inserted from above into the hollow portion of each of the two types of sampling pins 141. The tip of the positioning pin body 142A pushed out the gel sheet portion 4E inside the sampling pin 141. The pushed-out gel sheet portion 4E was placed on a slide glass, and its morphology was observed using a phase-contrast microscope.

[0091] Figure 33 is a photograph of a gel sheet portion collected with the first collection pin. Figure 34 is a photograph of a gel sheet portion collected with the second collection pin. Referring to Figures 33 and 34, both gel sheet portions have a round shape similar to the hollow portion of collection pin 141. The size (diameter) of the gel sheet portion collected with the first collection pin was 737 μm, and the size (diameter) of the gel sheet portion collected with the second collection pin was 1122 μm.

[0092] A similar test to the above was performed by forming gel sheets 4 of two different thicknesses (a first gel sheet with a thin thickness and a second gel sheet with a thicker thickness than the first gel sheet). Figure 35 is a photograph of a gel sheet portion taken from the first gel sheet. Figure 36 is a photograph of a gel sheet portion taken from the second gel sheet. Referring to Figures 35 and 36, the thickness of the gel sheet portion taken from the first gel sheet with a thickness of approximately 1000 µm was 1174 µm, and the thickness of the gel sheet portion taken from the second gel sheet with a thickness of approximately 2000 µm was 2286 µm. From the above, it was found that the thickness of the taken gel sheet portion depended on the thickness of the gel sheet.

[0093] From the above results, when collecting a gel sheet, it is possible to collect a gel sheet of any shape and size by controlling the dimensions of collection pin main body 141A and the thickness of the gel sheet.

[0094] The relationship between the inner diameter of the hollow portion of sampling pin body 141A and the outer diameter (diameter) of location pin body 142A is as follows: When a gel sheet portion collected with sampling pin body 141A having an inner diameter of approximately 0.7 mm was pushed out with location pin body 142A having an outer diameter of 0.5 mm, the gel sheet portion could be stably pushed out. However, when a gel sheet portion collected with sampling pin body 141A having an inner diameter of approximately 1.1 mm was pushed out with location pin body 142A having an outer diameter of 0.5 mm, it was difficult to stably push out the gel sheet portion. For this reason, in order to stably push out the gel sheet, it is preferable that the outer diameter of location pin body 142A be approximately 0.2 mm smaller than the inner diameter of sampling pin body 141A.

[0095] The features described in the above-described embodiments (examples) may be applied in appropriate combinations within the scope of technical compatibility.

[0096] The embodiments disclosed herein should be considered to be illustrative in all respects and not restrictive. The scope of the present invention is defined by the claims, not by the above description, and is intended to include all modifications within the meaning and scope of the claims.

[0097] Various aspects of the present disclosure are summarized below as appendices.

[0098] (Appendix 1) A hollow sampling pin capable of cutting and sampling the material to be treated; a positioning pin that can be inserted into the sampling pin, that contacts the material to be treated cut by pressing the sampling pin inside the sampling pin, and that presses the material to be treated to position the material to the outside of the sampling pin; a fixing holder to which the sampling pin is fixed, The processing target material is a collection and placement member in which a gel for supporting the target material to be collected is formed into a sheet-like gel sheet.

[0099] (Appendix 2) 2. The collection and placement member according to claim 1, wherein the placement pin can be inserted through the inside of the collection pin while being moved vertically by a first drive unit.

[0100] (Appendix 3) the first drive unit includes a first motor, a rotatable disk member attached to the first motor, and a link member having a first end fixed to a part of the outer periphery of the disk member and extending in the vertical direction; A collection and placement member as described in Appendix 2, wherein a second end portion opposite the first end portion in the extension direction of the link member moves vertically as the first motor rotates the disc member, thereby causing the second end portion to move the placement pin vertically.

[0101] (Appendix 4) 4. The collection and positioning member of claim 3, further comprising a positioning pin holder fixing portion fixed to the positioning pin between the second end and the positioning pin.

[0102] (Appendix 5) The positioning pin includes a positioning pin body and a positioning pin holder that holds the positioning pin body, 5. The collection and positioning member according to any one of claims 1 to 4, wherein a spring is provided in an area inside the positioning pin holder adjacent to the positioning pin body.

[0103] (Appendix 6) Further, a second drive unit is provided which can move the fixing holder in the up and down direction, The second drive unit includes a second motor and a vertically extending screw that extends in the vertical direction and has the second motor attached to one end thereof. The collection and placement member according to any one of appendices 1 to 5, including a screw connection portion that connects the vertically extending screw and the fixing holder and is slidable along the extension direction of the vertically extending screw.

[0104] (Appendix 7) A collection and placement member according to any one of appendices 1 to 6, A collection and placement device comprising a holding table that holds the material to be treated that is cut out by the collection pin.

[0105] (Appendix 8) Preparing a collection and placement member according to any one of appendices 1 to 7; A collection and placement method including the steps of: lowering the tip of the collection pin to cut out the material to be treated that is positioned below the collection pin and collect it in the collection pin; and lowering the tip of the placement pin to contact and press the material to be treated inside the collection pin, thereby placing the material to be treated outside the collection pin.

[0106] (Appendix 9) a step of forming a gel sheet on a film to support the object to be collected; a step of puncturing the gel sheet on the film with a hollow collection pin, and cutting out a gel sheet portion that is a part of the gel sheet, so that the gel sheet portion collected by the collection pin is pushed out from inside the collection pin by being pressed by a placement pin and placed outside the collection pin, In the step of placing, the placement pin is inserted through the interior of the collection pin.

[0107] (Appendix 10) 10. The collection and placement method of claim 9, wherein in the placing step, after the gel sheet portion is cut out by the collection pin to include the target object, the collection pin penetrates the film while holding the gel sheet portion inside.

[0108] (Appendix 11) 11. The collection and placement method of claim 9 or 10, wherein the object is embedded in the gel.

[0109] (Appendix 12) 12. The method for collecting and placing a sample according to any one of claims 9 to 11, wherein the gel is a gel of a biocompatible gel raw material.

[0110] (Appendix 13) 13. The collection and placement method according to any one of appendices 9 to 12, wherein the object is an isolated cell or a cell aggregate.

[0111] (Appendix 14) 14. The collection and placement method of claim 13, further comprising the step of recovering the object by dissolving the gel.

[0112] (Appendix 15) the target substance is any factor selected from the group consisting of a protein as a growth factor, a protein as an enzyme, a peptide, a compound, and a particle; the compound is any one selected from the group consisting of an adrenergic β1 receptor agonist / antagonist, a Rho kinase inhibitor, and a Wnt signaling pathway-related inhibitor; 13. The collection and placement method according to any one of appendices 9 to 12, wherein the particles are any one selected from the group consisting of plastics, lipid bilayers, and polysaccharides.

[0113] (Appendix 16) 16. The collection and placement method of claim 15, wherein the factor is released slowly after it is released from the collection pin. [Explanation of symbols]

[0114] 1, 5 slide glass, 2 film, 3 spacer, 3A through hole, 4 gel sheet, 4A gel raw material, 4B sample, 4C flow sample, 4D gel, 4E, 4H gel sheet portion, 4G factor, 6 gel plate, 7I application needle, 8 plate, 9A container, 10, 10A gel dissolving agent, 12 high viscosity solution, 15 concentration gradient, 21 sheet fixing member, 24 collection needle portion, 100 collection placement device, 101 XY stage for sample, 101A stage penetration portion, 102 XY stage for container, 104 collection placement member, 104A collection placement mechanism, 106 observation optical system, 141 collection pin, 141A collection pin main body, 141B collection pin holder, 142 placement pin, 142A placement pin main body, 142B placement pin holder, 142C spring, 143 Fixed holder, 143f, 146f front, 143b back, 144 first drive part, 144A first motor, 144B disc member, 144C link member, 144D1, 144D2 joint, 145 positioning pin holder fixing part, 145a first part, 145b second part, 146 second drive part, 146A movable holder, 146B second motor, 146C vertically extending screw, 146D screw connection part, 146E slide groove, FXD fixing part.

Claims

1. A sampling pin having a hollow shape capable of cutting and sampling the material to be treated; a positioning pin that can be inserted into the sampling pin, contacts the material to be treated that has been cut out by pressing the sampling pin, and presses the material to be treated, thereby positioning the material to the outside of the sampling pin; a fixing holder to which the sampling pin is fixed; The collection and placement member, wherein the processing target material is a gel sheet in which a gel for supporting the target material to be collected is formed into a sheet shape.

2. The collection and positioning member according to claim 1 , wherein the positioning pin is capable of being inserted through the inside of the collection pin while being moved in the vertical direction by a first drive unit.

3. the first drive unit includes a first motor, a rotatable disk member attached to the first motor, and a link member having a first end fixed to a portion of an outer periphery of the disk member and extending in a vertical direction; The collection and placement member of claim 2, wherein a second end opposite the first end in the extension direction of the link member moves vertically as the first motor rotates the disc member, thereby causing the second end to move the placement pin vertically.

4. The collection and positioning member of claim 3 , further comprising a locating pin holder fixing portion fixed to the locating pin between the second end and the locating pin.

5. The positioning pin includes a positioning pin body and a positioning pin holder that holds the positioning pin body, The collection and positioning member according to claim 1 or 2, wherein a spring is provided in an area inside the positioning pin holder adjacent to the positioning pin body.

6. Further, a second drive unit capable of moving the fixing holder in a vertical direction is provided, The second drive unit includes a second motor and a vertically extending screw that extends in the vertical direction and has one end to which the second motor is attached. The collection and placement member according to claim 1 or 2, further comprising a screw connection portion that connects the vertically extending screw and the fixing holder and is slidable along the extension direction of the vertically extending screw.

7. The collection arrangement member according to claim 1 ; A sampling and placement device comprising a holding table that holds the material to be treated that is cut out by the sampling pin.

8. Providing a collection arrangement member according to claim 1 or 2; A method for collecting and placing the material to be treated, comprising the steps of: lowering a tip of the collection pin to cut out the material to be treated that is positioned below the collection pin and collect it within the collection pin; and lowering the tip of the placement pin to contact and press the material to be treated within the collection pin, thereby placing the material to be treated outside the collection pin.

9. A step of forming a gel for supporting an object to be collected in a sheet shape on a film and providing a gel sheet; a step of puncturing the gel sheet on the film with a hollow collection pin, and cutting out a gel sheet portion that is a part of the gel sheet, so that the gel sheet portion collected by the collection pin is pushed out from inside the collection pin by being pressed by a placement pin and placed outside the collection pin, A collection and placement method, wherein in the placing step, the placement pin is inserted through the inside of the collection pin.

10. The collection and placement method according to claim 9, wherein in the placing step, after the gel sheet portion is cut out to include the object by the collection pin, the collection pin penetrates the film while holding the gel sheet portion inside.

11. The method of claim 9 or 10, wherein the object is embedded in the gel.

12. The method for collecting and placing a tissue according to claim 9 or 10, wherein the gel is a gel of a biocompatible gel material.

13. 11. The method of claim 9 or 10, wherein the object is an isolated cell or a clump of cells.

14. The method of claim 13, further comprising retrieving the object by dissolving the gel.

15. The target substance is any factor selected from the group consisting of a protein as a growth factor, a protein as an enzyme, a peptide, a compound, and a particle; The compound is adrenaline β 1 The compound is any one selected from the group consisting of a receptor agonist / antagonist, a Rho kinase inhibitor, and a Wnt signaling pathway-related inhibitor; The method for collecting and placing a sample according to claim 9 or 10, wherein the particles are any one selected from the group consisting of plastics, lipid bilayer membranes, and polysaccharides.

16. 16. The collection and deployment method of claim 15, wherein the factor is released slowly after it is released from the collection pin.

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