Collection arrangement member, collection arrangement device, and collection arrangement method

Abstract

The present invention comprises: a collection pin (141) having a hollow shape and capable of cutting and collecting a to-be-processed material; an arrangement pin (142) that contacts, inside the collection pin (141), the to-be-processed material in the collection pin (141), and presses the to-be-processed material to arrange the to-be-processed material outside the collection pin (141); and a fixing holder (143) to which the collection pin (141) is fixed. The to-be-processed material is a gel sheet in which a gel for supporting an object to be collected is formed in a sheet shape.

Description

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

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

[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 cells. From the viewpoint of appropriately collecting only a desired number of 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 technique for forming droplets that protrude outward from the tip of a coating needle by using a coating needle whose outer periphery is surrounded by an outer cylinder and 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.

[0006] International Publication No. 2017 / 061387 Japanese Patent Application Laid-Open No. 2020-015016

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

[0008] Furthermore, 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.

[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.

[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.

[0015] 6 is a schematic front view of the inside of the collecting and placing device according to the present embodiment; FIG. 7 is a front view of a collecting and placing mechanism constituting the collecting and placing member of FIG. 1, as seen from the negative side in the Y direction; FIG. 8 is a side view of the collecting and placing mechanism of FIG. 2, as seen from the negative side in the X direction; FIG. 9 is a front view of the entire collecting and placing member of FIG. 1, as seen from the negative side in the Y direction; FIG. 10 is a side view of the collecting and placing member of FIG. 4, as seen from the negative side in the X direction; FIG. 11 is a front view of the collecting and placing member of FIG. 4, as seen from the negative side in the Y direction, showing the operation of the collecting and placing member of FIG. 4 in a step of cutting and collecting material to be treated; FIG. 6 is a side view of the operation of the collecting and placing mechanism of FIG. 2, as seen from the negative side in the Y direction, showing the operation of the first step of releasing and placing material to be treated; FIG. 8 is a front view of the operation of the second step following FIG. 8, as seen from the negative side in the Y direction; FIG. 9 is a front view of the operation of the third step following FIG. 10, as seen from the negative side in the X direction; FIG. 11 is a side view of the operation of FIG. 10, as seen from the negative side in the X direction; and FIG. 12 is a front view of the operation of the fourth step following FIG. 10, as seen from the negative side in the Y direction. 19 is a front view, seen from the negative side in the Y direction, of the operation in the fifth step following FIG. 12 . FIG. 20 is a schematic cross-sectional view showing a first step of the collection and placement method according to the present embodiment. FIG. 21 is a schematic cross-sectional view showing a second step of the collection and placement method according to the present embodiment. FIG. 22 is a schematic cross-sectional view showing a third step of the collection and placement method according to the present embodiment. FIG. 23 is a schematic cross-sectional view showing a fourth step of the collection and placement method according to the present embodiment. FIG. 24 is a schematic cross-sectional view showing a fifth step of the collection and placement method according to the present embodiment. FIG. 25 is a schematic enlarged perspective view showing the plate included in FIG. 18 and the gel plate thereon. FIG. 26 is a schematic cross-sectional view showing a sixth step of the collection and placement method according to the present embodiment. FIG. 27 is a schematic enlarged cross-sectional view showing a portion where the collection pin cuts off the gel plate immediately after FIG. 20 . FIG. 28 is a schematic enlarged cross-sectional view showing a seventh step of the collection and placement method according to the present embodiment. FIG. 29 is a schematic enlarged cross-sectional view showing an eighth step of the collection and placement method according to the present embodiment. FIG. 29 is a schematic cross-sectional view showing an example of a step in which the gel is dissolved from the gel sheet portion to recover only cells. FIG. 22 is a schematic enlarged cross-sectional view in which the sample in FIG. 22 is replaced with a factor. FIG. 29 is a schematic enlarged cross-sectional view showing a step in which the factor contained in the gel is gradually released or eluted from the gel sheet portion in FIG. 25 . FIG. 29 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. 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.21 ; FIG. 22 ; FIG. 23 ; FIG. 24 ; FIG. 25 ; FIG. 26 ; FIG. 27 ; FIG. 28 ; FIG. 29 ; FIG. 30 ; FIG. 31 ; FIG. 32 ; FIG. 33 ; FIG. 34 ; FIG. 35 ; FIG. 36 ; FIG. 37 ; FIG. 38 ; FIG. 39 ; FIG. 40 ; FIG. 41 ; FIG. 42 ; FIG. 43 ; FIG. 44 ; FIG. 45 ; FIG. 46 ; FIG. 47 ; FIG. 48 ; FIG. 49 ; FIG. 50 ; FIG. 51 ; FIG. 52 ; FIG. 53 ; FIG. 54 ; FIG. 55 ; FIG. 56 ; FIG. 57 ; FIG. 58 ; FIG. 59 ; FIG. 60 ;

[0016] This embodiment will now be described with reference to the drawings. (Collection and placement member) First, the features of this embodiment will be briefly described. Referring to FIG. 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 fixing 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.

[0017] 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 (described later). The sampling and placement device 100 mainly comprises a processing chamber, a sample XY stage 101 disposed within the processing chamber, a container XY stage 102, and a sampling and placement member 104. While the sampling and placement device 100 in FIG. 1 has only one sampling and placement member 104, it may include multiple members.

[0018] 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 101A that penetrates at least a portion of the sample XY stage 101 is formed. It is preferable that the gel plate 6 be fixed at a position overlapping the stage penetration 101A.

[0019] The container XY stage 102 is disposed below the sample XY stage 101. The container XY stage 102 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 container XY stage 102. The guide unit is slidably connected to a guide rail provided on the bottom surface of the processing chamber. The container XY stage 102 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 arranged in this order from top to bottom in the Z direction.

[0020] 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.

[0021] Although not shown, a control unit is installed outside the processing chamber. The control unit includes a monitor, a control computer, and an operation panel. In the control unit, a command value for the movement speed of the needle for sampling from the operation panel is input and stored in the storage device of the control computer. For example, during a sample sampling operation, the command value for the speed 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 pins 141 and 142 up and down at a predetermined speed. This allows 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 the 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 speed of the sampling operation may be calculated depending on the material, thickness, and sampling method of the specified gel sheet.

[0022] (Configuration of the sampling and placement mechanism) FIG. 2 is a front view of the sampling and placement mechanism constituting the sampling and placement member of FIG. 1, as viewed from the negative side in the Y direction. FIG. 3 is a side view of the sampling and placement mechanism of FIG. 2, as viewed from the negative side in the X direction. Referring to FIGS. 2 and 3, the sampling and placement member 104 of FIG. 1 includes a sampling and placement mechanism 104A, which is a part of the sampling and placement member 104. The sampling and placement mechanism 104A mainly includes a sampling pin 141, a placement 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 placement 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 placement pin 142 are formed of, for example, a metal such as stainless steel or plastic.

[0023] The sampling pin 141 comprises 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 circumferential surface of the upper portion 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 circumferential surface of the sampling pin body 141A by press-fitting. Alternatively, the sampling pin holder 141B is adhered to the outer circumferential surface of the sampling pin body 141A with adhesive 141C. 2 and 3, the sampling pin holder 141B may be arranged such that the top surface thereof is aligned with the XY plane, but the bottom surface thereof is inclined relative to the XY plane, so that the outer periphery has 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.

[0024] 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.

[0025] 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.

[0026] The sampling pin body 141A presses the material to be processed by descending, cutting out a portion of the material, and stores the cut 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 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 stored inside the sampling pin body 141A out of the sampling pin 141. The positioning pin body 142A can move up and down within 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 up and down movement of the positioning pin body 142A is controlled by a first drive unit 144, described below. This allows the cut material to be placed in a desired position.

[0027] A spring 142C is installed in the area of ​​the inside of the positioning pin holder 142B adjacent to the positioning pin 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 body 142A. The spring 142C absorbs the Z-direction force applied to the positioning pin holder 142B when the positioning pin body 142A collides with the workpiece 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.

[0028] The fixing holder 143 can hold the collection pin 141. The fixing holder 143 is disposed above the collection pin 141 and behind the positioning pin 142 in FIG. 2 . The fixing holder 143 has, for example, a plate shape (rectangular parallelepiped shape), but is not limited to this. The collection pin 141 is fixed to the fixing holder 143. Specifically, the top surface of the collection 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 collection 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 collection pin holder 141 to the fixing holder 143.

[0029] A first drive unit 144 is provided on the 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 drive unit 144 includes a first motor 144A, a disk member 144B, and a link member 144C. The first drive unit 144 is attached to a positioning pin holder fixing portion 145.

[0030] 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 rear main surface 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 front main surface opposite to 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.

[0031] The link member 144C is installed on the front side of the disk 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 disk shape of the disk 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 disk member 144B. At least a portion of the first motor 144A may be disposed within the fixing holder 143. The disk 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 portion of the outer periphery of the disk member 144B.

[0032] A second end (the lower end in FIG. 2 ) of the link member 144C, which is opposite to the first end in the extension direction (Z direction), is fixed to the positioning pin holder fixing portion 145 so as to contact the first portion 145a of the positioning pin holder fixing portion 145. The second end may be capable of contacting 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.

[0033] To achieve this operation, a 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 contact 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.

[0034] (Overall Configuration of the Collection and Placement Component) Figure 4 is a front view of the entire collection and placement component of Figure 1, viewed from the negative side in the Y direction. Figure 5 is a side view of the collection and placement component of Figure 4, viewed from the negative side in the X direction. With reference to Figures 4 and 5, the collection and placement component 104 further includes a second drive unit 146 in addition to the collection and placement mechanism 104A of Figures 2 and 3. The second drive unit 146 is disposed on the rear side of the fixed holder 143 of Figures 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.

[0035] The second drive unit 146 is a slide mechanism that slides the collection and placement mechanism 104A, including the fixed holder 143, in the Z direction. In addition to the movable holder 146A, the second drive unit 146 also includes a second motor 146B, a vertically extending screw 146C, and a screw connection 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 fixed holder 143. The vertically extending screw 146C may be housed in a groove extending in the Z direction formed on the front surface 146f of the movable holder 146A. 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.

[0036] 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.

[0037] (Collection Process) Figure 6 is a front view of the collection and placement member of Figure 4 in the process of cutting and collecting the material to be treated, as viewed from the negative Y-axis direction. Figure 7 is a side view of the operation of Figure 6, as viewed from the negative X-axis direction. Referring to Figures 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, vertically. It is preferable that the front surface 146f of the movable holder 146A, which faces the rear surface 143b of the fixing holder 143, be formed with a slide groove 146E (see Figure 6) extending along the Z-axis 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 (i.e., dug from the front side to the back side of the paper in FIG. 6). In this case, the screw connection portion 146D is disposed so that at least a portion thereof fits within the slide groove 146E.

[0038] 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 treated that is positioned below the tip. The cut-out portion of the material to be treated is stored in the hollow portion of the sampling pin body 141A.

[0039] (Placement Process) Figure 8 is a front view of the collection and placement mechanism of Figure 2, seen from the negative side in the Y direction, showing the operation of the first process of releasing and placing the material to be treated. Referring to Figure 8, the first end of link member 144C is fixed to a portion of the outer periphery of disc member 144B by joint 144D1. The second end of link member 144C is fixed to placement pin holder fixing portion 145 by joint 144D2. In the initial state of Figure 8, the first end of link member 144C is located at the top of disc member 144B. Therefore, the placement pin holder fixing portion 145 and placement pin 142 connected to link member 144C are located at the top in the Z direction. The tip (bottom) of placement pin main body 142A is located above the bottom of collection pin main body 141A. In this state, the bottom of collection pin main body 141A becomes the bottom of the entire assembly, and this portion can be used to cut out the material to be treated.

[0040] 9 is a front view, seen from the negative side in the Y direction, of the operation in the second step following FIG. 8. 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. This positions the first end of the link member 144C at the center position of the disk member 144B in the Z direction. Therefore, the link member 144C tilts slightly with respect to the Z direction, and lowers the positioning pin holder fixing portion 145 and the positioning pin 142 connected thereto, as indicated by the arrow M2 in the figure.

[0041] FIG. 10 is a front view of the operation in the third step following FIG. 9 , as viewed from the negative Y-axis direction. FIG. 11 is a side view of the operation in FIG. 10 , as 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. This causes the tip of the positioning pin body 142A to contact and press downward against the target material stored in the sampling pin body 141A, thereby ejecting it from the sampling pin body 141A and positioning it in the desired position.

[0042] 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 is further rotated, 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 the initial state in Fig. 8 is returned to as shown in Fig. 13.

[0043] 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 does not press downward, positioning pin holder fixing portion 145 rises due to the biasing force of the spring.

[0044] (Details of the Object Collection and Arrangement Method) FIG. 14 is a schematic cross-sectional view showing a first step of the object collection and arrangement method according to this embodiment. Referring to FIG. 14 , a film 2 is placed on, for example, a glass slide 1. As an example, the glass slide 1 is a thin glass plate material approximately 1.2 mm thick. The film 2 is formed thinly from a soft material so that it can be easily broken by puncturing with a needle. As an example, the film 2 is made of polyvinylidene chloride approximately 10 μm thick. That is, it is preferable to use a thin film for food packaging or a thin film of silicone rubber 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 the top (plan view) of 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.

[0045] Next, a fluid sample 4C, which includes one or more target samples 4B contained in 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.

[0046] 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.

[0047] 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.

[0048] 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. The gel 4D is solidified, but 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.

[0049] FIG. 17 is a schematic cross-sectional view showing the 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.

[0050] Fig. 18 is a schematic cross-sectional view showing the fifth step of the collection and placement method according to this embodiment. Fig. 19 is a schematic enlarged perspective view showing the plate shown in Fig. 18 and the gel plate thereon. Referring to Figs. 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 (Figs. 6 and 7).

[0051] 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. Due to 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.

[0052] The multiple containers 9A may be spaced apart from one another, for example, in eight rows in the depth direction and twelve rows in the width direction in FIG. 19 , for a total of 96 containers 9A. The planar shape of the containers 9A may be any 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.

[0053] 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 in 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 in position.

[0054] The plate 8 on which the container 9A serving as the culture container is formed is made of any material selected from the group consisting of polystyrene, polypropylene, polycarbonate, and polyethylene terephthalate.

[0055] 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, descends 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 positioning pin body 142A is positioned at the top, and the bottom of the sampling pin body 141A is positioned below the bottom of the positioning 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 positioning pin body 142A) be large enough to accommodate the cut gel sheet portion.

[0056] 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 is preferably composed of a gel 4D and a sample 4B embedded therein. In this way, the sample 4B does not flow 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.

[0057] After the gel sheet portion 4E has been cut off by the sampling pin body 141A, the sampling pin body 141A continues to descend while holding the gel sheet portion 4E inside (the 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.

[0058] 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 the process shown in 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, for example, to the state shown in FIGS. 10 and 11 above. 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 interior (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.

[0059] 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, and not the film 2, be 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 be collected within the container 9A, and not the film 2.

[0060] 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.

[0061] (Material) Gel 4D is a gel formed from biocompatible gel raw material 4A. Biocompatible here means that it does not cause inflammation or other toxicity when contacted with biological tissues such as cells, and has low invasiveness to cells. This reduces damage to cells (specimen 4B) caused by the gel. Gel raw material 4A is preferably transparent or translucent, allowing for observation of the position of embedded specimen 4B. To satisfy the above requirements, gel raw material 4A is 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).

[0062] The sample 4B as the object may be, for example, an isolated cell or a cell aggregate. Here, "isolated" refers to both a case in which only one sample 4B is contained within the gel sheet portion 4E, as shown in Figures 21 to 23, and a case in which multiple samples 4B are contained, each spaced apart, 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.

[0063] 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 prevents 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, the formation of an aggregate is acceptable.

[0064] 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 ５A gel sheet portion 4E containing fibroblasts at 0.1 cells / mL within a gel 4D is contained within the device. 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 within 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 can be used as the gel solubilizer 10.

[0065] 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 shown in 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 parenthesized symbol 4G 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.

[0066] 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.

[0067] Figure 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 Figure 25. Referring to Figure 26, a gel solubilizer is supplied to the gel sheet portion 4H, which embeds factor 4G and is placed in the container 9A of Figure 25, in the same manner as in Figure 24. The gel solubilizer dissolves the gel 4D, resulting in a state where the gel solubilizer 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 factor 4G. A concentration difference of factor 4G occurs between the gel sheet portion 4H embedding factor 4G and the solution 10A, which does not contain 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 factor 4G. That is, after the factor 4G is released from the sampling pin 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 even if the gel 4D does not dissolve.

[0068] (Effects) 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 comes into contact with the material to be processed cut out by pressing the collection pin 141 inside the collection pin 141, and by pressing the material to be processed, places 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 which a gel 4D is formed into a sheet shape and supports a sample 4B, which is the object to be collected.

[0069] 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 sample 4B within the material to be treated, which are a concern 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 of the sample 4B, etc., and improves the positional accuracy and reproducibility of the placement process of the sample 4B, etc.

[0070] 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 141. This allows the target object to be cut out with high stability and reproducibility.

[0071] In the collection and placement member 104, the placement pin 142 can be inserted into the collection pin 141 while being moved up and down 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.

[0072] 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, 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 vertical reciprocating motion 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.

[0073] 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.

[0074] In the collection and placement member 104, the placement pin 142 includes a placement pin main body 142A and a placement pin holder 142B that holds the placement pin main body 142A. A spring 142C is installed in an area inside the placement pin holder 142B adjacent to the placement pin main body 142A. This provides the effect of stably positioning the gel sheet portion 4E, etc., in the desired position using the placement pin main body 142A.

[0075] 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 having the second motor 146B attached to one end (e.g., the upper end) thereof, and a screw connector 146D connecting the vertically extending screw 146C to the fixing holder 143 and slidable along the extension direction of the vertically extending screw 146C. The second drive unit 146 converts the rotation of the second motor 146B and the vertically extending screw 146C into vertical reciprocating motion 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, enabling the desired portion of the gel sheet 4 to be cut out with high positional accuracy.

[0076] The sampling and placement device 100 according to the present disclosure includes the sampling and placement member 104 described above and a holding stage (sample XY stage 101) that holds the material to be treated that is cut out by the sampling pin 141. The sampling and placement device 100 configured in this manner achieves the effects provided by the sampling and placement member 104 described above.

[0077] 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 contacted and pressed against the material inside the collection pin 141, and the material to be treated is placed outside the collection pin 141.

[0078] 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 sample 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 sample 4B, etc., and improves the positional accuracy and reproducibility of the placement process for the sample 4B, etc.

[0079] In the collection and placement method according to the present disclosure, a gel supporting the object 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 the collection pin 141 by pressure from a placement pin 142 and placed outside the collection pin 141. In the step of placing the gel sheet portions outside the collection pin 141, the placement pin 142 is inserted through the interior of the collection pin 141.

[0080] 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 the object, such as the sample 4B, which can occur when the gel sheet portions 4E, 4H are ejected from the collection pin 141 using liquid pressure or the like. 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.

[0081] In the step of disposing the target outside the collection pin 141, the collection pin 141 cuts out the gel sheet portions 4E, 4H so as to include the target, and then the collection 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 collection 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.

[0082] (Modification) FIG. 27 is a schematic enlarged cross-sectional view showing a first step of supplying a high-viscosity solution into a container in a modification. FIG. 28 is a schematic enlarged cross-sectional view showing a second step of supplying a high-viscosity solution into a container in a modification. FIG. 29 is a schematic enlarged cross-sectional view showing a third step of supplying a high-viscosity solution into a container in a modification. 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 (e.g., the bottom surface). The coating needle 7I may be solid or hollow. Alternatively, a droplet dispensing mechanism using a commonly known dispenser may be used to dispense 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 .

[0083] 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.

[0084] Figure 30 is a schematic enlarged cross-sectional view corresponding to Figure 21 of a modified example in which a high-viscosity solution is supplied. Figure 31 is a schematic enlarged cross-sectional view corresponding to Figure 22 of a modified example in which a high-viscosity solution is supplied. Figure 32 is a schematic enlarged cross-sectional view corresponding to Figure 23 of a modified example in which a high-viscosity solution is supplied. With reference to Figures 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 Figures 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 Figures 21 to 23.

[0085] 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, etc.

[0086] 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.

[0087] 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.

[0088] Next, as shown in Figure 21, the collection pin 141 was lowered almost vertically from above, penetrating the gel sheet 4 and the film 2 to collect the gel sheet portion 4E. Two types of collection and placement members 104 were used: one with a collection pin body 141A (hereinafter referred to as the first collection pin) having an inner diameter of 0.7 mm and an outer diameter of 1 mm, and the other with a collection pin body 141A (hereinafter referred to as the second collection 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.

[0089] 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.

[0090] 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.

[0091] A similar test to the above was performed by forming gel sheets 4 of two different thicknesses (a first gel sheet having a thinner thickness and a second gel sheet having a thicker thickness than the first gel sheet). FIG. 35 is a photograph of a gel sheet portion taken from the first gel sheet. FIG. 36 is a photograph of a gel sheet portion taken from the second gel sheet. Referring to FIGS. 35 and 36, the thickness of the gel sheet portion taken from the first gel sheet having a thickness of approximately 1000 μm was 1174 μm, and the thickness of the gel sheet portion taken from the second gel sheet having 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.

[0092] From the above results, it is possible to collect gel sheets of any shape and size by controlling the dimensions of collection pin body 141A and the thickness of the gel sheet.

[0093] The relationship between the inner diameter of the hollow portion of the sampling pin body 141A and the outer diameter (diameter) of the positioning pin body 142A is as follows: When a gel sheet portion collected with a sampling pin body 141A having an inner diameter of approximately 0.7 mm was pushed out with a positioning 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 a sampling pin body 141A having an inner diameter of approximately 1.1 mm was pushed out with a positioning pin body 142A having an outer diameter of 0.5 mm, it was difficult to stably push out the gel sheet portion. Therefore, in order to stably push out the gel sheet, it is preferable that the outer diameter of the positioning pin body 142A be approximately 0.2 mm smaller than the inner diameter of the sampling pin body 141A.

[0094] The features described in the above-described embodiments (examples) may be applied in appropriate combinations within the scope of technical compatibility.

[0095] 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.

[0096] Various aspects of the present disclosure are summarized below as appendices. (Appendix 1) A collection and placement component comprising: a hollow collection pin capable of cutting and collecting material to be processed; a placement pin that can be inserted into the collection pin and that contacts the material to be processed cut out by pressing the collection pin inside the collection pin and presses the material to be processed, thereby placing the material to be processed outside the collection pin; and a fixing holder to which the collection pin is fixed, wherein the material to be processed is a gel sheet formed into a sheet-like gel that supports the material to be collected.

[0097] (Supplementary Note 2) The collection and positioning member according to Supplementary Note 1, wherein the positioning pin can be inserted through the inside of the collection pin while being moved in the up and down direction by a first drive unit.

[0098] (Appendix 3) The collection and placement member described in Appendix 2, wherein the first drive unit includes a first motor, a rotatable disc member attached to the first motor, and a link member having a first end fixed to a portion of the outer periphery of the disc member and extending in an up-down direction, and a second end of the link member opposite the first end in the extension direction moves in an up-down direction as the disc member is rotated by the first motor, thereby causing the second end to move the placement pin in an up-down direction.

[0099] (Supplementary Note 4) The collection and positioning member according to Supplementary Note 3, further comprising a positioning pin holder fixing portion fixed to the positioning pin between the second end and the positioning pin.

[0100] (Appendix 5) The collection and placement member described in any one of Appendices 1 to 4, wherein the placement pin includes a placement pin body and a placement pin holder that holds the placement pin body, and a spring is installed in an area inside the placement pin holder adjacent to the placement pin body.

[0101] (Appendix 6) A collection and placement member as described in any one of Appendices 1 to 5, further comprising a second drive unit capable of moving the fixing holder in an up and down direction, wherein the second drive unit includes a second motor, a vertically extending screw that extends in an up and down direction and has the second motor attached to one end thereof, and 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.

[0102] (Supplementary Note 7) A collection and placement device comprising: the collection and placement member according to any one of Supplementary Notes 1 to 6; and a holding table that holds the material to be treated that is cut out by the collection pin.

[0103] (Appendix 8) A collection and placement method comprising the steps of: preparing a collection and placement member as described in any one of Appendices 1 to 7; and lowering the tip of the collection pin to cut out the material to be treated that is placed below the collection pin and place it inside the collection pin, and then 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.

[0104] (Appendix 9) A collection and placement method comprising: a step of forming a gel that supports an object to be collected into a sheet shape on a film, and providing a gel sheet; and a step of puncturing the gel sheet on the film with a collection pin having a hollow shape, 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, wherein in the placing step, the placement pin is inserted inside the collection pin.

[0105] (Appendix 10) The collection and placement method described in Appendix 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.

[0106] (Appendix 11) The collection and placement method according to appendix 9 or 10, wherein the object is embedded in the gel.

[0107] (Appendix 12) The collection and placement method according to any one of Appendices 9 to 11, wherein the gel is a gel of a biocompatible gel raw material.

[0108] (Appendix 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.

[0109] (Appendix 14) The collection and placement method according to Appendix 13, further comprising the step of recovering the object by dissolving the gel.

[0110] (Appendix 15) The collection and placement method according to any one of Appendices 9 to 12, wherein the target object is a 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 a compound selected from the group consisting of an adrenergic β1 receptor agonist / antagonist, a Rho kinase inhibitor, and a Wnt signaling pathway-related inhibitor; and the particle is a compound selected from the group consisting of a plastic, a lipid bilayer membrane, and a polysaccharide.

[0111] (Appendix 16) The collection and placement method of appendix 15, wherein the factor is released gradually after being released from the collection pin.

[0112] 1, 5 slide glass, 2 film, 3 spacer, 3A through hole, 4 gel sheet, 4A gel raw material, 4B sample, 4C fluid 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 sample XY stage, 101A stage penetration portion, 102 container XY stage, 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 Fixing holder, 143f, 146f front, 143b back, 144 first drive unit, 144A first motor, 144B disc member, 144C link member, 144D1, 144D2 joint, 145 positioning pin holder fixing portion, 145a first portion, 145b second portion, 146 second drive unit, 146A movable holder, 146B second motor, 146C vertically extending screw, 146D screw connection portion, 146E slide groove, FXD fixing portion.

Claims

1. A collection and placement member comprising: a collection pin having a hollow shape capable of cutting out and collecting a material to be treated; a placement pin that can be inserted into the inside of the collection pin and that contacts the material to be treated cut out by pressing the collection pin within the collection pin and presses the material to be treated, thereby placing the material to be treated outside the collection pin; and a fixing holder to which the collection pin is fixed, wherein the material to be treated is a gel sheet made of a gel formed into a sheet shape to support the object to be collected.

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.

3. The collection and positioning member described in claim 2, wherein the first drive unit includes a first motor, a rotatable disc member attached to the first motor, and a link member having a first end fixed to a portion of the outer periphery of the disc member and extending in an upward and downward direction, and a second end of the link member opposite the first end in the extension direction moves in an upward and downward direction as the disc member is rotated by the first motor, thereby causing the second end to move the positioning pin in an upward and downward direction.

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

5. A collection and placement member as described in claim 1 or 2, wherein the placement pin includes a placement pin body and a placement pin holder that holds the placement pin body, and a spring is installed in an area of ​​the interior of the placement pin holder adjacent to the placement pin body.

6. A collection and placement member as described in claim 1 or 2, further comprising a second drive unit capable of moving the fixing holder in an up and down direction, the second drive unit including a second motor, a vertically extending screw extending in an up and down direction and having the second motor attached to one end, and a screw connection portion connecting the vertically extending screw and the fixing holder, and capable of sliding along the extension direction of the vertically extending screw.

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

8. A collection and placement method comprising the steps of: preparing a collection and placement member as described in claim 1 or 2; and lowering a tip of the collection pin to cut out the material to be treated that is located below the collection pin and place 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 collection and placement method comprising the steps of: forming a gel supporting an object to be collected into a sheet-like form on a film, and providing a gel sheet; and puncturing the gel sheet on the film with a hollow collection pin, and cutting out a portion of the gel sheet that is a part of the gel sheet, so that the portion of the gel sheet collected by the collection pin is pushed out from within the collection pin by pressure from a placement pin and placed outside the collection pin, 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. The method of claim 9 or 10, wherein the object is an isolated cell or a clump of cells.

14. The collection and placement method of claim 13, further comprising the step of retrieving said object by dissolving said gel.

15. The collection and placement method described in claim 9 or 10, wherein the target object is any factor selected from the group consisting of proteins as growth factors, proteins as enzymes, peptides, compounds and particles, the compound is any one selected from the group consisting of adrenergic β1 receptor agonists / antagonists, Rho kinase inhibitors and Wnt signaling pathway-related inhibitors, and the particle is any one selected from the group consisting of plastics, lipid bilayer membranes and polysaccharides.

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

Images