Gel sheet chip and method for manufacturing the same

Abstract

To provide a gel sheet chip and a sampling set which are used for sampling excellent in stability, reproductivity, and location accuracy, and a manufacturing method of the gel sheet chip.SOLUTION: A gel sheet chip 6 includes a film 2, a frame 3, a gel 4D, and samples 4B to be collected. The frame 3 is placed on the film 2 and has a through hole 3A. The gel 4D is placed on the film 2 in the through hole 3A. The samples 4B to be collected are supported by the gel 4D.SELECTED DRAWING: Figure 7

Description

Technical Field

[0001] The present disclosure relates to a gel sheet chip, a method for manufacturing the same, and a sample collection set.

Background Art

[0002] In recent years, techniques for selecting target cells and cell populations by analyzing single cells have been rapidly developing. As techniques for selecting target cells, cell selection methods such as the limiting dilution method, cell sorter, microfluidic channel, and colony pickup method have attracted attention.

[0003] In addition, as a technique for handling a small number of cells, a printing technique for handling a small number of droplets has attracted attention. The printing techniques include an inkjet method, a dispenser method, and a coating method.

[0004] However, there are cases where it is preferable to collect only one cell. From the viewpoint of appropriately collecting only one arbitrary cell, a technique for precisely extracting even smaller droplets with higher accuracy than the above-described printing techniques is required. From this viewpoint, a method using a manipulator disclosed in Japanese Patent Application Laid-Open No. 2008-152044 (Patent Document 1) and a method for cutting and collecting a biological sample such as a cell using a sampling needle disclosed in International Publication No. 2017 / 061387 (Patent Document 2) have been developed.

Prior Art Documents

Patent Documents

[0005]

Patent Document 1

Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

[0006] In Japanese Patent Publication No. 2008-152044, a solution containing the cells to be collected is drawn into the hollow part of a tube. Since the cells to be collected are contained in the solution, if the solution is discharged into a container, the cells will disperse inside the container. This presents a challenge in that it is difficult to collect samples with high stability and reproducibility.

[0007] In International Publication No. 2017 / 061387, the gel comes into contact with the cell specimen. The cut cell specimen is temporarily placed inside the collection needle. Then, liquid is injected into the collection needle, and the pressure causes the cell specimen inside the needle to be expelled from the needle. However, the gel is not fixed and supported around the cells. Therefore, especially with cells that are not adherent to the gel, the cell specimen may break down and disperse when expelled from the collection needle. There is also a risk of damaging the cells by piercing them with the collection needle. The position of the cells is not fixed by the gel, making it difficult to collect cells with high precision and reproducibility.

[0008] This disclosure has been made in view of the above-mentioned problems. The object of this disclosure is to provide a gel sheet chip, a sample collection set, and a method for manufacturing the gel sheet chip, which are used for sample collection with high stability, reproducibility, and positional accuracy. [Means for solving the problem]

[0009] A gel sheet chip to which a sample to be collected is fixed, according to this disclosure, has the sample to be collected fixed to it. The gel sheet chip comprises a film, a frame, a gel, and a sample. The frame is placed on the film and a through-hole is formed therein. The gel is placed on the film within the through-hole. The sample to be collected is supported by the gel.

[0010] A method for manufacturing a gel sheet chip on which a sample to be collected is fixed, according to this disclosure, involves placing a frame with through-holes on a film. A fluid sample containing the sample to be collected is supplied into the through-holes on the film. The thickness of the fluid sample is made uniform by sandwiching the structure, which consists of the film, frame, and fluid sample, between flat plates placed on the upper and lower sides. The fluid sample is then gelled.

[0011] A sample collection set for fixing a sample to be collected according to this disclosure comprises a gel material, a film, and a frame. The gel material is capable of supporting the sample to be collected by forming a gel. The film is capable of supporting the gel. The frame is capable of being placed on the film and has through holes. [Effects of the Invention]

[0012] According to this disclosure, it is possible to provide a gel sheet chip, a sample collection set, and a method for manufacturing the gel sheet chip that enable high-precision sample collection with high stability, reproducibility, and positional accuracy. [Brief explanation of the drawing]

[0013] [Figure 1] This is a schematic front view of the inside of the sample collection device according to this embodiment. [Figure 2] Figure 1 is a schematic diagram illustrating the sampling mechanism of the sample collection device shown. [Figure 3] This is a schematic diagram illustrating the configuration of the sampling needle holder in the sampling mechanism shown in Figure 2. [Figure 4] This is a schematic perspective view of the film and frame constituting the gel sheet chip according to this embodiment. [Figure 5] Figure 4 is a schematic perspective view showing the frame placed on the film. [Figure 6] This is a schematic cross-sectional view of the portion along the line VI-VI in Figure 5. [Figure 7] This is a schematic perspective view of a gel sheet chip according to a first example of this embodiment. [Figure 8]It is a schematic cross-sectional view showing the first step of the method for manufacturing a gel sheet chip according to the first example of the present embodiment. [Figure 9] It is a schematic cross-sectional view showing the second step of the method for manufacturing a gel sheet chip according to the first example of the present embodiment. [Figure 10] It is a schematic cross-sectional view showing the third step of the method for manufacturing a gel sheet chip according to the first example of the present embodiment. [Figure 11] It is a schematic cross-sectional view showing the fourth step of the method for manufacturing a gel sheet chip according to the first example of the present embodiment. [Figure 12] It is a schematic perspective view of a gel sheet chip according to the second example of the present embodiment. [Figure 13] It is a schematic cross-sectional view showing the first step of the method for manufacturing a gel sheet chip according to the second example of the present embodiment. [Figure 14] It is a schematic cross-sectional view showing the second step of the method for manufacturing a gel sheet chip according to the second example of the present embodiment. [Figure 15] It is a schematic cross-sectional view showing the third step of the method for manufacturing a gel sheet chip according to the second example of the present embodiment. [Figure 16] It is a schematic cross-sectional view showing the fourth step of the method for manufacturing a gel sheet chip according to the second example of the present embodiment. [Figure 17] It is a schematic perspective view of a gel sheet chip according to the third example of the present embodiment. [Figure 18] It is a schematic cross-sectional view showing the first step of the supporting method in an example where a gel supports a sample by embedding. [Figure 19] It is a schematic cross-sectional view showing the second step of the supporting method in an example where a gel supports a sample by embedding. [Figure 20] It is a schematic perspective view showing the first step of the supporting method in an example where a gel supports a sample by adhesion. [Figure 21] It is a schematic perspective view showing the second step of the supporting method in an example where a gel supports a sample by adhesion. [Figure 22] It is a schematic cross-sectional view of a portion along line XXII-XXII in FIG. 21. [Figure 23]This is a schematic cross-sectional view showing the third step of the support method in an example where a gel supports a sample by adhesion. [Figure 24] This is a schematic cross-sectional view showing the fourth step of the support method in an example where a gel supports a sample by adhesion. [Figure 25] This is a schematic diagram showing a modified version of Figure 24. [Figure 26] This is a schematic diagram showing a sample collection set for fixing a sample to be collected, according to this embodiment. [Figure 27] This is a schematic diagram showing the first step of a sample collection method using a gel sheet chip in which the gel supports the sample by embedding it. [Figure 28] This is a schematic diagram showing the second step of a sample collection method using a gel sheet chip in which the gel supports the sample by embedding it. [Figure 29] This is a schematic diagram showing the third step of a sample collection method using a gel sheet chip in which the gel supports the sample by embedding it. [Figure 30] This is a schematic diagram showing the first step of a sample collection method using a gel sheet chip in which the gel adheres to the sample. [Figure 31] This is a schematic diagram showing the second step of a sample collection method using a gel sheet chip in which the gel adheres to the sample. [Figure 32] This is a schematic diagram showing the third step of a sample collection method using a gel sheet chip in which the gel adheres to the sample. [Figure 33] This is a schematic cross-sectional view showing the first example of a process in which the gel is dissolved from the gel sheet and only the cells are recovered. [Figure 34] This is a schematic cross-sectional view showing a second example of a process in which the gel is dissolved from the gel sheet and only the cells are recovered. [Modes for carrying out the invention]

[0014] This embodiment will be described below with reference to the drawings.

[0015] First, the features of this embodiment will be briefly described. As shown in Figure 7, the gel sheet chip 6 comprises a film 2, a frame 3, a gel 4D, and a sample 4B to be collected. The frame 3 is placed on the film 2, and a through hole 3A is formed. The gel 4D is placed on the film 2 so as to be in contact with the wall surface of the through hole 3A. The sample 4B is supported by the gel 4D. The embodiment will now be described in detail.

[0016] (Overall configuration of the sample collection device) Figure 1 is a schematic front view of the interior of the sample collection device according to Embodiment 1. For the sake of explanation, the X, Y, and Z directions are introduced. Referring to Figure 1, the sample collection device 100 is a device for collecting a sample from a gel sheet chip 6, which will be described later. The sample collection device 100 mainly comprises a processing chamber, an XY stage 101 for the sample located inside the processing chamber, an XY stage 102 for the container, and a collection mechanism 104. Although the sample collection device 100 in Figure 1 has only one collection mechanism 104, it may include multiple such mechanisms.

[0017] The sample XY stage 101 (sample stage) is movable horizontally, i.e., along the XY direction (X direction and Y direction). Specifically, for example, a guide is installed on the lower surface of the sample XY stage 101. This guide is slidably connected to a guide rail installed on the bottom surface of the processing chamber. The upper surface of the sample XY stage 101 is a mounting surface on which a gel sheet chip 6 can be fixed. At least a portion of the sample XY stage 101 has a stage penetration portion 101A that penetrates it. It is preferable that the gel sheet chip 6 is fixed in a position that overlaps with the stage penetration portion 101A.

[0018] Below the sample XY stage 101, a container XY stage 102 (container stage) is positioned. The container XY stage 102 is movable horizontally, i.e., along the XY direction (X and Y directions). Specifically, for example, a guide is installed on the underside of the container XY stage 102. This guide is slidably connected to a guide rail installed on the bottom surface of the processing chamber. The container XY stage 102 fixes a container 9A that can house a portion of the gel sheet chip 6. As a result, in Figure 1, the sampling mechanism 104, gel sheet chip 6, and container 9A are arranged in the order from top to bottom in the Z direction.

[0019] The sampling mechanism 104 and the observation optical system 106 are connected to a member that is movable in the Z direction, such as a Z-axis table. In other words, the sampling mechanism 104 and the observation optical system 106 are held within the sample collection device 100 so as to be movable in the Z direction. The observation optical system 106 observes and measures the position of the sample to be collected contained in the gel sheet chip 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 sheet chip 6 by the observation optical system 106 may be performed using visible light. However, observation of the gel sheet chip 6 is not limited to visible light; it may also be performed using infrared light, X-rays, ultrasound, etc., and depending on the material of the gel sheet chip 6, it may be possible to observe the gel sheet chip 6 using magnetism. The gel sheet chip 6 observed by means other than visible light does not need to be transparent or translucent, and may be opaque.

[0020] Although not shown in the diagram, 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, command values ​​for the operating speed of the needle for sampling, obtained from the operation panel, are input and stored in the control computer's storage device. For example, during sample collection, the command value for the speed is read from the storage device and sent to the control program of the sampling mechanism 104. Based on this command value, the control program of the sampling mechanism 104 determines the rotational speed of the servo motor 41 (described later) included in the sampling mechanism 104 and moves the sampling needle unit 24 up and down at a predetermined speed. This performs the sampling operation of the sampling mechanism 104. If the control computer is communicating with an upstream control system, the above command values ​​may be received from that control system. In addition, parameters corresponding to the material of the gel sheet from which the sample is collected may be stored in the storage device, and the command value for the sampling operation speed may be calculated according to the specified gel sheet material, thickness, and sampling method.

[0021] (Configuration of the sampling mechanism and sampling needle holder) Figure 2 is a schematic diagram showing the sampling mechanism of the sample collection device shown in Figure 1. Figure 3 is a schematic diagram illustrating the configuration of the sampling needle holder in the sampling mechanism shown in Figure 2. The sampling mechanism 104 described above will be explained in more detail with reference to Figures 2 and 3.

[0022] The sampling mechanism 104 in Figure 1 includes a sampling needle holder 7H containing a sampling needle pin 7B. The sampling mechanism 104 includes a first drive unit 40. The sampling needle holder 7H is detachably connected to the first drive unit 40. Any structure can be used for connecting the sampling needle holder 7H to the first drive unit 40.

[0023] Any configuration can be adopted for the first drive unit 40. For example, as shown in Figure 2, the first drive unit 40 includes a servo motor 41, a cam 43, a bearing 44, a cam connecting plate 45, and a movable part 46. The servo motor 41 is installed such that its rotation axis extends in the direction along the Z direction as shown in Figure 1. The cam 43 is connected to the rotation axis of the servo motor 41. The cam 43 is rotatable about the rotation axis of the servo motor 41.

[0024] The cam 43 includes a central part connected to the rotation axis of the servo motor 41 and a flange part connected to one end of the central part. The upper surface of the flange part (the surface on the servo motor 41 side) is the cam surface. This cam surface is formed in an annular shape along the outer circumference of the central part. Furthermore, as shown in Figure 2, the cam surface is formed in a slope shape so that the distance from the bottom surface of the flange part varies. For example, the cam surface includes an upper flat region where the distance from the bottom surface of the flange part is greatest, a lower flat region located at a distance from the upper flat region and where the distance from the bottom surface of the flange part is smallest, and a slope part connecting the upper flat region and the lower flat region.

[0025] A bearing 44 is positioned so as to be in contact with the cam surface of the cam 43. As shown in Figure 2, the bearing 44 is positioned in a specific direction (to the right of the servo motor 41) when viewed from the cam 43. The bearing 44 maintains contact with the cam surface when the cam 43 rotates due to the rotation of the servo motor 41's rotation axis. A cam connecting plate 45 is connected to this bearing 44. In the cam connecting plate 45, one end connected to the bearing 44 and the other end opposite to it are fixed to a movable part 46. A collection needle holder housing is connected to the movable part 46. The collection needle holder 7H described above is housed in this collection needle holder housing. For example, a first connecting member such as a magnet may be placed on the surface of the collection needle holder 7H facing the collection needle holder housing. Alternatively, a second connecting member such as a magnet or magnetic material may be placed on the collection needle holder housing side to fix the first connecting member on the collection needle holder 7H side.

[0026] A fixing pin is installed on the movable part 46. Another fixing pin is installed on the frame that holds the servo motor 41. A spring is installed connecting these two fixing pins. Due to this spring, the movable part 46 is subjected to a downward tensile force. This tensile force from the spring acts on the bearing 44 via the movable part 46 and the cam connecting plate 45. This tensile force from the spring maintains the bearing 44 in a state of being pressed against the cam surface of the cam 43.

[0027] Furthermore, the movable part 46 and the collection needle holder housing are connected to a linear guide installed on the frame. The linear guide is positioned to extend in the Z direction. Therefore, the movable part 46 and the collection needle holder housing (including the pin 7B and collection needle holder 7H) are movable along the Z direction.

[0028] Figure 3 is a schematic diagram illustrating the configuration of the sampling needle holder in the sampling mechanism shown in Figure 2. Referring to Figure 3, the sampling needle holder 7H is capable of gripping the central sampling equipment consisting of a syringe 7A and a pin 7B. The pin 7B has a hollow, or hollow, section 7C inside. The pin 7B can cut and penetrate the target object by descending.

[0029] Pin 7B is fixed to syringe 7A. In the collection needle holder 7H, syringe 7A is fixed by an upper syringe fixing part 7D and a lower syringe fixing part 7E. A syringe cap 7CP is attached to the upper side of syringe 7A. A tube 7G is connected to the upper side of syringe cap 7CP. Gas or liquid is injected into syringe 7A from tube 7G through syringe cap 7CP. As a result, a portion of the cut gel sheet tip 6, which is housed in the hollow part 7C of pin 7B, is pressed downwards and discharged from pin 7B.

[0030] The sampling needle holder 7H houses the above components within the holder shell 7F. However, parts of the pin 7B and tube 7G may protrude outside the holder shell 7F.

[0031] (Gel sheet chip and method for manufacturing the same) Figure 4 is a schematic perspective view of the film and frame constituting the gel sheet chip according to this embodiment. Figure 5 is a schematic perspective view showing the frame placed on the film of Figure 4. Figure 6 is a schematic cross-sectional view of the portion along the line VI-VI in Figure 5. Referring to Figures 4 to 6, the gel sheet chip according to this embodiment comprises a film 2 and a frame 3.

[0032] Film 2 is, for example, a thin film having a rectangular shape in plan view. Film 2 may be formed from any material selected from the group consisting of polyethylene, polyvinylidene chloride, and polyvinyl chloride. Alternatively, film 2 may be formed from either polydimethylsiloxane or butyl rubber. The thickness of film 2 is preferably 1 mm or less. In particular, the thickness of film 2 is preferably 10 μm or more and 100 μm or less. Also from the above viewpoint, the maximum elongation (ultimate elongation) of film 2 is preferably 100% or more and 1300% or less. By using such material and size, film 2 is soft and brittle enough to be easily torn by the small pressure caused by the descent of the pin described later, but the parts other than the puncture site do not tear significantly and can support the gel sheet or the like on top even after penetration. In other words, film 2 makes the puncture site even easier to tear when pin 7B is descent, while preventing damage to parts other than the puncture site. Furthermore, in order to visually confirm the position of sample 4B through film 2, film 2 is preferably transparent or translucent, suitable for observation by an optical system.

[0033] Preferably, frame 3 has substantially the same shape and size as film 2 in plan view, for example. That is, frame 3 has a rectangular outer shape, similar to film 2. As shown in Figures 5 and 6, frame 3 is placed on film 2. A portion of frame 3 (in plan view), i.e., the central part in plan view, is penetrated. In other words, a through-hole 3A is formed in frame 3. The through-hole 3A is hollow, and no material constituting frame 3 is placed inside. The through-hole 3A may be rectangular, similar to the outer shape of frame 3. The through-hole 3A may be similar in shape to the outer shape of frame 3. The center of the through-hole 3A may coincide with the center of the entire frame 3. The size of the through-hole 3A is determined according to the size of the gel sheet 4 (see Figure 7), which is determined according to the type and amount of sample 4B to be embedded.

[0034] The frame 3 is preferably made of a material that is highly rigid and capable of supporting the film 2 and gel sheet 4 (see Figure 7) from below. Specifically, the frame 3 may be formed from any of the group selected from polystyrene or other plastic materials, glass, metal, silicone rubber-based materials, and nonwoven materials.

[0035] Figure 7 is a schematic perspective view of a gel sheet chip according to the first example of this embodiment. Referring to Figure 7, the gel sheet chip 6 includes a gel sheet 4 in addition to the film 2 and frame 3 described above. The gel sheet 4 contains a sample 4B and a gel 4D. The sample 4B is cells to be collected. In Figure 7, multiple cells, which are samples 4B, are arranged one by one with space between them. The sample 4B is supported by the solidified gel 4D. The gel 4D is solidified but soft enough to be easily broken by pin puncture. The statement that the sample 4B is supported by the gel 4D means, for example, that the sample 4B is embedded so that it is covered and buried by the hard gel 4D, and the sample 4B does not flow. Therefore, in Figure 7, the sample 4B is arranged so that it is contained inside the gel 4D.

[0036] In Figure 7, the gel sheet chip 6 has a gel 4D thickness that is approximately the same as the frame 3 thickness T1. Here, "approximately the same thickness" does not only mean that the dimensions are exactly the same, but also includes cases where the difference between the average thickness of gel 4D and the average thickness of frame 3 is within 10% of the average thickness of frame 3.

[0037] The gel 4D, which supports the sample 4B, for example by embedding, is placed on the film 2 within the through-hole 3A of the frame 3. Preferably, the gel 4D is placed in contact with the four walls (side surfaces as inner walls) of the through-hole 3A. At least a portion (only a portion) of the four walls of the through-hole 3A may be in contact with the walls of the gel 4D. The entire four walls of the through-hole 3A may be in contact with the walls of the gel 4D. As the frame 3 is placed on the film 2, the bottom of the through-hole 3A is covered by the film 2. A container-like region is formed by the film 2 covering the through-hole 3A and the walls of the through-hole 3A. The gel 4D is housed within this container-like region so as to be in contact with both the walls of the through-hole 3A and the film 2. This forms the gel sheet chip 6.

[0038] Gel 4D is a gel made from a biocompatible gel material. Biocompatibility means that it does not exhibit toxicity such as inflammation even when it comes into contact with living tissues such as cells, and has low invasiveness to cells. This suppresses damage to cells (sample 4B) caused by the gel. The biocompatibility of a material can be confirmed, for example, by observing that the number of living cells does not decrease when cells are cultured on the material and stained to distinguish between living and dead cells, or when inflammatory markers expressed by cells or contents eluted from cells are measured. Alternatively, the biocompatibility of a material can be confirmed by observing that the expression of inflammatory markers does not increase, or that contents do not elute from cells. Furthermore, the gel material 4A is preferably transparent or translucent from the viewpoint of allowing observation of the position of the embedded sample 4B. In addition, it is preferable that gel 4D is made of a material that has adhesive properties to cells. Adhesion means that after cells are seeded on the surface of gel 4D and left to stand for more than 4 hours, the cells adhere to the surface of gel 4D.

[0039] From the standpoint of satisfying the above, the 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).

[0040] In the gel sheet chip 6, it is preferable that the film 2 and the frame 3 are attached using an adhesive such as cyanoacrylate. This is because cyanoacrylate is less invasive to the cells, which are sample 4B.

[0041] Next, the manufacturing method of the gel sheet chip 6 shown in Figure 7 will be described using Figures 8 to 11. Figure 8 is a schematic cross-sectional view showing the first step of the manufacturing method of the gel sheet chip according to the first example of this embodiment. Referring to Figure 8, a film 2 is placed on a flat plate, such as a glass slide 1. A frame 3 with through holes 3A formed therein is attached to the film 2, for example, with the adhesive described above.

[0042] Next, a fluid sample 4C, which contains one or more samples 4B within a fluid gel raw material 4A capable of forming a gel, is supplied into the through-hole 3A on the film 2. In Figure 13, multiple samples 4B are arranged one by one, spaced apart from each other.

[0043] Figure 9 is a schematic cross-sectional view showing the second step of the manufacturing method for a gel sheet chip according to the first example of this embodiment. Referring to Figure 9, a film 2 is attached to the lower surface of the slide glass 5. The slide glass 5 may be of the same material and size as the slide glass 1 in Figure 8. That is, the slide glass 5 in Figure 9 is a flat plate with a nearly uniform overall thickness. The film 2 on the upper side of the fluid sample 4C in Figure 5 may be of the same material and size as the film 2 in Figure 4.

[0044] Figure 10 is a schematic cross-sectional view showing the third step of the manufacturing method for a gel sheet chip according to the first example of this embodiment. Referring to Figure 10, a slide glass 5 with film 2 attached is placed on a frame 3. The film 2 attached to the slide glass 5 is in contact with the fluid sample 4C and the frame 3. The slide glass 5 and film 2 press down on the fluid sample 4C and the frame 3. As a result, the lower structure in Figure 10, consisting of film 2 (attached to slide glass 1), frame 3, and fluid sample 4C, is sandwiched between the slide glass 1 with film 2 attached below it and the slide glass 5 with film 2 attached above it. Thus, from bottom to top, slide glass 1, film 2, frame 3 and fluid sample 4C, film 2, and slide glass 5 are stacked in that order. In addition, the thickness of the fluid sample 4C is made uniform by the contact of the slide glass 5 and the flat plate made of film 2 from above. Here, uniform thickness does not mean that the thickness of the fluid sample 4C is completely the same throughout, but rather that the difference between the maximum and minimum thickness of the fluid sample 4C is within 1% of the average thickness of the fluid sample 4C.

[0045] Next, while the structure is left to stand in the above state, if, for example, the gel material 4A is a material that solidifies depending on temperature, the entire stacked structure is cooled to, for example, 4°C. Due to the cooling, the gel material 4A constituting the fluid sample 4C becomes a solidified gel 4D, and a gel sheet 4 is formed in the through-hole 3A with the sample 4B embedded inside the gel 4D. In this way, if the gel material 4A is a material that gels with temperature, the above structure is left to stand in an optimal temperature environment. Although the gel 4D is solidified, it is soft enough to be easily broken by pin puncture.

[0046] Figure 11 is a schematic cross-sectional view showing the fourth step of the manufacturing method for a gel sheet chip according to the first example of this embodiment. Referring to Figure 11, after the gel sheet 4 is formed, the film 2 and slide glass 5 placed on top of the gel sheet 4 are removed, and the slide glass 1 is also removed. This forms a gel sheet chip 6 comprising the film 2, the frame 3, and the gel sheet 4 placed on the film 2. In other words, Figure 11 shows a cross-sectional view of the gel sheet chip 6 in the form of Figure 7.

[0047] Figure 12 is a schematic perspective view of a gel sheet chip according to a second example of this embodiment. Referring to Figure 12, the gel sheet chip 6 has a largely similar configuration to the gel sheet chip 6 of the first example in Figure 7. Therefore, the same reference numerals are used for the same components as in Figure 7, and the description is not repeated as long as the function, etc., is the same. However, in the gel sheet chip 6 of Figure 12, the gel 4D is thinner than the frame 3. In particular, here the difference between the average thickness of the gel 4D and the average thickness of the frame 3 exceeds 10% of the average thickness of the frame 3. For example, the average thickness of the gel 4D is half the average thickness of the frame 3. Also, the gel 4D (including the gel raw material 4A) in Figure 12 (including Figures 13 to 16) includes both cases, as described later, where the sample 4B is supported inside the gel 4D and where it is supported by adhering it to the outside of the gel 4D. From the perspective of including both of these cases, the sample 4B is not shown inside the gel sheet 4 in Figure 12. Combining the examples in Figure 7 and Figure 12, in this embodiment, the thickness of gel 4D is less than or equal to the thickness of frame 3.

[0048] Next, the manufacturing method of the gel sheet chip 6 shown in Figure 12 will be described using Figures 13 to 16. However, the explanation of parts that overlap with the steps in Figures 8 to 11 will not be repeated. Figure 13 is a schematic cross-sectional view showing the first step of the manufacturing method of the gel sheet chip according to the second example of this embodiment. Referring to Figure 13, the same process as in Figure 8 is basically performed. A fluid gel raw material 4A is supplied into the through hole 3A.

[0049] Figure 14 is a schematic cross-sectional view showing the second step of the method for manufacturing a gel sheet chip according to a second example of this embodiment. Referring to Figure 14, the process is basically the same as in Figure 9. However, in Figure 14, the thickness of the region inside the through hole 3A of the glass slide 5A is thicker than the thickness of the region outside the through hole 3A. Specifically, the difference in thickness between the region inside the through hole 3A and the other regions of the glass slide 5A is made equal to the difference between the thickness of the frame 3 and the thickness of the gel material 4A to be formed after compression.

[0050] Figure 15 is a schematic cross-sectional view showing the third step of the manufacturing method for a gel sheet chip according to a second example of this embodiment. Referring to Figure 15, the process is basically the same as in Figure 10. However, in Figure 15, a slide glass 5A with film 2 attached is placed on a frame 3. The slide glass 5A and film 2 press down on the gel material 4A and the frame 3. The slide glass 5A protrudes downward in the region that overlaps with the through hole 3A compared to the other regions. As a result, the protruding portion enters the through hole 3A and presses down on the gel material 4A. This makes the gel material 4A thinner than the frame 3.

[0051] Figure 16 is a schematic cross-sectional view showing the fourth step of the method for manufacturing a gel sheet chip according to a second example of this embodiment. Referring to Figure 16, the process is basically the same as in Figure 11. Figure 16 shows a cross-sectional view of the gel sheet chip 6 of Figure 11.

[0052] Figure 17 is a schematic perspective view of a gel sheet chip according to the third example of this embodiment. Referring to Figure 17, the gel sheet chip 6 has a largely similar configuration to the gel sheet chip 6 of the first example in Figure 7. Therefore, the same reference numerals are used for the same components as in Figure 7, and the explanation will not be repeated as long as the function, etc., is the same. However, in the gel sheet chip 6 of Figure 17, the thickness of the frame 3 is T2, and the sample 4B is in the form of a cell aggregate. That is, multiple samples 4B are gathered together to form a clump. In this respect, Figure 17 differs in configuration from Figure 7, in which the samples 4B are supported one by one on the gel 4D, separated from each other. The aggregate needs to be large enough to pass through the hollow-shaped part of the pin, which will be described later. That is, the aggregate needs to be smaller than the width in the direction intersecting the direction in which the hollow-shaped part extends. In Figure 17, as an example, three samples 4B are gathered in the aggregate, but the aggregate is not limited to this, and may consist of two or four or more samples 4B.

[0053] Next, the method of supporting sample 4B with gel 4D will be explained using Figures 18 to 25.

[0054] Figure 18 is a schematic cross-sectional view showing the first step of the support method in an example where the gel supports the sample by embedding. Figure 19 is a schematic cross-sectional view showing the second step of the support method in an example where the gel supports the sample by embedding. Referring to Figure 18, when forming a gel sheet chip 6 in which the sample 4B is supported so as to be embedded in the gel 4D, as in Figures 7 to 11 (Figures 12 to 16), the sample 4B is embedded in the gel raw material 4A, similar to Figure 8. Figure 18 shows the same process as in Figure 8, but the amount of fluid sample 4C supplied into the through-hole 3A on the film 2 is less than in Figure 8. Referring to Figure 19, the formed gel sheet chip 6 is broadly the same as in Figure 11, but the gel sheet 4 is formed thinner than in Figure 11. In other words, in Figure 19, the gel sheet 4 is formed thinner than the frame 3 shown in Figures 12 and 16. Figure 19 shows an example in which the gel sheet 4 is arranged so that the sample 4B is embedded in the gel 4D.

[0055] In the example where sample 4B is embedded in gel 4D, the first example shown in Figures 7 to 11 (where gel sheet 4 is approximately the same thickness as frame 3) may be applied, or the example shown in Figures 18 and 19 (where gel sheet 4 is thinner than frame 3) may be applied.

[0056] In examples where sample 4B is embedded and supported within gel 4D, as shown in Figures 18-19 (including Figures 7-11), the cells in sample 4B may be non-adherent or adherent to gel 4D. Conversely, gel 4D may or may not be made of a material that adheres to cells.

[0057] Figure 20 is a schematic perspective view showing the first step of a support method in an example where the gel supports the sample by adhesion. Referring to Figure 20, for example, a gel sheet chip is formed by the same method as in Figures 8 to 11 (except that the sample 4B is included in the gel raw material 4A), comprising only gel 4D without sample 4B. In Figure 20, the thickness of gel 4D is approximately equal to the thickness of frame 3. However, in Figure 20, a gel 4D thinner than frame 3 may be formed by the same method as in Figures 12 to 16. In Figure 20, it is preferable that gel 4D is made of a material that has adhesive properties to cells. Gel 4D is formed of, for example, collagen.

[0058] Figure 21 is a schematic perspective view showing the second step of the support method in an example where the gel supports the sample by adhesion. Figure 22 is a schematic cross-sectional view of the portion along the line XXII-XXII in Figure 21. Referring to Figures 21 and 22, another frame 3B is placed on top of the frame 3 of Figure 20. Preferably, the other frame 3B has the same shape, substantially the same size, and the same material as frame 3. Through holes 3C are formed in the other frame 3B. Preferably, the through holes 3C have the same shape and substantially the same size as through holes 3A.

[0059] Figure 23 is a schematic cross-sectional view showing the third step of the support method in an example where the gel supports the sample by adhesion. Referring to Figure 23, the sample 4B and the culture medium 20 in which it is suspended are supplied into the through-hole 3C of the frame 3B. Preferably, the culture medium 20 is supplied so as to fill the through-hole 3C. If the gel 4D is thinner than the frame 3, the sample 4B and a portion of the culture medium 20 may be positioned so as to enter the area within the through-hole 3A.

[0060] Figure 24 is a schematic cross-sectional view showing the fourth step of the support method in an example where the gel supports the sample by adhesion. Referring to Figure 24, the supplied sample 4B descends compared to the supply stage in Figure 23 and adheres to the upper surface of the gel 4D. This forms a gel sheet chip 6A comprising a film 2, a frame 3, another frame 3B, the gel 4D on the film 2, the sample 4B adhered to the gel 4D, and a culture medium 20 filling the through-holes 3C of the other frame 3B.

[0061] In the examples shown in Figures 18 to 19, as in the examples shown in Figures 20 to 24, a culture medium and buffer solution may be supplied into the through-hole 3A to prevent drying.

[0062] Figure 25 is a schematic diagram showing a modified version of Figure 24. Referring to Figure 25, for example, a gel 4D formed thinner than the frame 3, similar to Figures 12 or 16, is subjected to the same treatment as in Figures 23-24. In Figure 25, the sample 4B is supported on the upper surface of the gel 4D by adhesion. In Figure 25, the culture medium 20 is supplied so as to immerse the sample 4B. As a result, the through-hole 3A is filled with the gel 4D, the sample 4B, and the culture medium 20. Figure 25 has a similar configuration to Figure 24 in general. However, in Figure 25, there is no frame 3B, only the frame 3, and the through-hole 3A is filled with the gel 4D, the sample 4B, and the culture medium 20. Therefore, in Figure 25, a gel sheet chip 6B is formed comprising a film 2, a frame 3, the gel 4D on the film 2, the sample 4B adhered to the gel 4D in the through-hole 3A of the frame 3, and the culture medium 20 filling the through-hole 3A of the frame 3. Such a configuration is also acceptable.

[0063] As described above, in this embodiment, the statement that the sample 4B is supported by the gel 4D includes not only the example in which the sample 4B is embedded in the gel 4D as shown in Figure 11, but also the example in which the sample 4B is fixed to the gel 4D by adhering to the outer surface of the gel 4D as shown in Figures 24 and 25. In any of the above examples, the gel sheet 4 is formed by the gel 4D and the sample 4B.

[0064] Figure 26 is a schematic diagram showing a sample collection set for fixing a sample to be collected according to this embodiment. Referring to Figure 26, in this embodiment, the sample collection set 11 may be implemented in the form of a sample collection set 11 comprising the components used above, namely a gel raw material 4A (medicine) capable of supporting the sample 4B to be collected by forming a gel, a film 2 on which the gel can be placed, and a frame 3. The frame 3 can be placed on the film 2 and has through holes 3A formed therein. In other words, the gel raw material 4A, film 2 and frame 3 are not integrated into the sample collection set 11 but are included as separate components. In addition to the above, the sample collection set 11 may further include another frame 3B. The sample collection set 11 allows the user to support the sample using the fluid gel raw material 4A before it is fixed. The sample collection set 11 allows the user to arbitrarily create gel sheet chips 6, etc., with a high degree of freedom. Using the sample collection set 11, the above sample collection method can be easily carried out by the user.

[0065] (Sampling method) Figure 27 is a schematic diagram showing the first step of a sample collection method using a gel sheet chip in which the gel supports the sample by embedding. Referring to Figure 27, for example, the gel sheet chip 6 shown in Figures 7 and 11 is placed on the sample XY stage 101. At least a portion of the gel sheet 4 constituting the gel sheet chip 6 is positioned to overlap with the stage penetration portion 101A formed to penetrate the sample XY stage 101. In particular, the sample 4B to be collected from the gel sheet 4 is positioned to overlap with the stage penetration portion 101A.

[0066] Figure 28 is a schematic diagram showing the second step of a sample collection method using a gel sheet chip in which the gel supports the sample by embedding it. Referring to Figure 28, a sample collection needle, such as pin 7B, descends from the state shown in Figure 27. The position where pin 7B should be descended is the location of the target sample 4B to be collected, which can be determined from the image acquired using an observation optical system (such as the observation optical system 106 in Figure 4). At this time, the gel sheet 4 of the gel sheet chip 6 is punctured by pin 7B. That is, the punctured portion of the gel sheet 4 is damaged by the passage of the sharp tip of pin 7B. Pin 7B penetrates the gel sheet 4 while tearing through it. The portion of the gel sheet 4 that is cut off from the original gel sheet 4 by pin 7B is the gel sheet portion 4E. Preferably, the cut-off gel sheet portion 4E consists of the gel 4D and the sample 4B embedded inside it. The pin 7B has a hollow portion 7C formed so as to penetrate it along its extending direction. The cut-out gel sheet portion 4E is guided into the hollow-shaped portion 7C and housed within the hollow-shaped portion 7C.

[0067] From the above state, pin 7B continues to descend. Pin 7B penetrates the portion of the gel sheet 4E that is being cut off, and the portion of the film 2 located directly beneath the gel sheet 4E. As a result, the film 2 is damaged in the same way as the gel sheet 4.

[0068] Figure 29 is a schematic diagram showing the third step of a sample collection method using a gel sheet chip in which the gel supports the sample by embedding. Referring to Figure 29, the pin 7B continues to descend from the state in which the pin 7B has penetrated the gel sheet 4 and film 2 as in Figure 28. At this time, the pin 7B discharges the gel sheet portion 4E from within the hollow portion 7C. The discharged gel sheet portion 4E is collected in the container 9A.

[0069] The gel sheet portion 4E is discharged from the hollow section 7C as follows: When the pin 7B is lowered to a position where at least the tip reaches the inside of the container 9A, air pressure is supplied to the hollow section 7C. The inside of the hollow section 7C under the pin 7B is pressurized. As a result, the gel sheet portion 4E inside the hollow section 7C is pressed downward and pushed out of the hollow section 7C.

[0070] Figure 30 is a schematic diagram showing the first step of a sample collection method using a gel sheet chip in which the gel adheres to the sample. Referring to Figure 30, for example, the gel sheet chip 6A shown in Figure 24 is placed on the sample XY stage 101, and the same process as in Figure 27 is basically performed.

[0071] Figure 31 is a schematic diagram showing the second step of a sample collection method using a gel sheet chip in which the gel adheres to the sample. Referring to Figure 31, the process is basically the same as in Figure 28.

[0072] Figure 32 is a schematic diagram showing the third step of a sample collection method using a gel sheet chip in which the gel adheres to the sample. Referring to Figure 32, the process is basically the same as in Figure 29.

[0073] (Effectiveness of sample collection methods) Since suspended cells cannot adhere to a substrate, fixation for collection is difficult. Furthermore, when collecting samples such as cells cultured in solid culture media within a thin-film culture device by aspiration, there is a concern that collection accuracy may decrease.

[0074] Therefore, in this embodiment, as described above (Figures 27 to 32), in the sample collection method, a portion of the gel sheet 4 is cut off by puncturing it with a pin 7B having a hollow shape portion 7C, and the gel sheet portion 4E is housed inside the hollow shape portion 7C. At this time, the pin 7B penetrates the gel sheet 4 and the portion of the film 2 directly beneath it. The gel sheet portion 4E housed inside the hollow shape portion 7C due to the penetration is collected in the container 9A.

[0075] According to the sample collection method described above, the sample 4B is supported by the gel 4D of the gel sheet 4, preventing it from flowing relative to the gel 4D. This suppresses the collapse and dissipation of the sample 4B when the gel sheet portion 4E is placed in the hollow-shaped portion 7C and during collection. Because the sample 4B is supported by the gel 4D, the possibility of the sample 4B moving in an unintended direction and damaging the sample 4B during puncture is reduced. Therefore, sample collection with high stability, reproducibility, and positional accuracy can be performed. For this reason, cells that are sufficiently supported and fixed can be collected with high accuracy compared to aspirating samples cultured in solid medium.

[0076] Furthermore, if a solid needle is used for contact, especially when collecting animal cells, there is a risk of damaging the cells due to contact with the needle. In addition, in methods of separating cells using liquid flow such as cell sorters and microfluidics, the high speed of the liquid flow can damage the separated cells, which is a problem. However, in this embodiment, a method is used in which the gel sheet portion 4E is housed within the hollow portion 7C of the pin 7B. When the gel sheet 4 is cut and collected so that it contains only one cell, if the cell is supported by embedding, for example, in the sheet member (gel), then the cell that is embedded in the gel and supported is cut out. This reduces the possibility of directly puncturing and damaging the cell.

[0077] Furthermore, as shown in Figures 29 and 32, in the above sample collection method, in the step where the gel sheet portion 4E is collected in the collection unit, only the gel sheet portion 4E may be collected in the collection unit. Generally, when cutting out a sheet member to which cells are attached, the sheet member is also cut out together with the cells. As a result, the material of the sheet member may be mixed with the collected cells, which can lead to problems in accurately analyzing the cells. However, according to this embodiment, unlike the sheet member, the film 2, which is a separate component from the gel sheet portion 4E, is not collected in the collection unit. Therefore, the possibility that the material of the film 2 may be mixed with the collected sample 4B and affect the accuracy of the analysis of sample 4B can be eliminated.

[0078] (Effects and Benefits) In this embodiment, a gel sheet chip on which a sample to be collected is fixed is disclosed. The gel sheet chip 6 comprises a film 2, a frame 3, a gel 4D, and a sample 4B to be collected. The frame 3 is placed on the film 2 and a through hole 3A is formed therein. The gel 4D is placed on the film 2 within the through hole 3A. The sample 4B to be collected is supported by the gel 4D.

[0079] The sample 4B is supported by the gel 4D within the gel sheet 4. Therefore, even if the gel sheet 4 is pierced by the pin 7B shown in Figures 27 to 32, the sample 4B will not flow relative to the gel 4D. This suppresses the collapse and dissipation of the sample 4B when the gel sheet portion 4E is placed in the hollow-shaped portion 7C and during sampling. Because the sample 4B is supported by the gel 4D, the possibility of the sample 4B moving in an unintended direction and damaging the sample 4B during puncture is reduced. Thus, sample collection with high stability, reproducibility, and positional accuracy can be performed. If the gel 4D and film 2 are made of a soft and easily torn material, they will easily break when pierced by the pin 7B. Therefore, the process of cutting out a part of the gel sheet 4 is not difficult.

[0080] As shown in Figures 10 and 15, a flat plate (glass slide 5) is placed on the frame 3. Therefore, the downward pressure on the glass slide 5 controls the thickness of the gel material 4A within the through-hole 3A. In other words, the frame 3 controls the thickness of the gel material 4A. This allows the sample 4B to be embedded without protruding or separating from the gel 4D. The sample 4B is securely supported (embedded or adhered) to the gel 4D, preventing displacement or dissipation of the sample 4B. Thus, highly stable, reproducible, and positionally accurate sample collection is possible.

[0081] Furthermore, it is preferable that the gel 4D is in contact with the inner wall surface that forms the through-hole 3A of the frame 3. This has the effect of preventing the dissipation and displacement of the sample 4B supported by the gel 4D (the sample 4B is fixed in the desired position with high precision).

[0082] If the gel sheet chip were to consist only of the film 2 and gel sheet 4 without a frame 3, handling the fluid sample 4C would be difficult. However, in this embodiment, the gel sheet chip 6 is equipped with a frame 3. By bringing the fluid sample 4C, surrounded by the frame 3, into contact with the inner wall surface, handling the fluid sample 4C becomes easier. As a result, displacement of the sample 4B contained in the gel 4D (gel sheet 4) is prevented.

[0083] In the gel sheet chip 6 described above, the thickness of the gel 4D may be less than or equal to the thickness of the frame 3. In other words, the thickness of the gel 4D may be approximately the same as the thickness of the frame 3 (see Figures 7 and 11). The gel 4D may also be thinner than the frame 3 (see Figures 12 and 16). Due to the presence of the frame 3, the thickness of the gel 4D can be freely controlled according to the shape and dimensions of the slide glass 5, 5A during the manufacturing of the gel sheet chip 6. By freely controlling the thickness of the gel 4D, a gel sheet portion 4E of any size can be cut out (see Figures 29 and 32).

[0084] In the gel sheet chip 6 described above, the sample 4B may be placed on the upper surface of the gel 4D so as to adhere to the gel 4D. This has the following advantages compared to, for example, the case where the sample 4B is placed so as to be contained within the gel 4D: If the sample 4B is cells, a mixture of cells with adhesive ability and cells without adhesive ability can be adhered to the gel 4D before the sample collection operation, thereby selectively separating only the cells that adhere to the gel 4D.

[0085] In the gel sheet chip 6 described above, another frame 3B may be further provided on top of the frame 3 so as to surround the sample 4B on the upper surface of the gel 4D. This allows for the formation of a gel sheet chip 6 in which, for example, the gel 4D is filled into the through-hole 3A of the lower frame 3, the sample 4B is placed in the through-hole 3C of the frame 3B above it, and the sample 4B adheres to the upper surface of the gel 4D.

[0086] Conversely, in the gel sheet chip 6 described above, the sample 4B may be placed inside the gel 4D within the through-hole 3A. This allows the sample 4B to be fixed regardless of whether or not it adheres to the gel 4D, and any sample 4B can be separated from the gel 4D.

[0087] In the gel sheet tip 6 described above, the sample 4B may be either cells or cell aggregates. For example, if multiple samples 4B gather in the gel sheet 4 to form an aggregate, there is a risk of puncturing and damaging the sample 4B with the pin 7B, especially if the aggregate is large. Damage to the sample 4B may make it difficult to analyze, for example, a cross-section of the tissue. Therefore, it is preferable that the multiple samples 4B be dispersed so that they are far apart from each other. This suppresses damage to the sample 4B due to puncture. However, if the aggregate is particularly small and can pass through the hollow section 7C, and the possibility of the sample 4B being punctured by the pin 7B is small, this configuration is also acceptable.

[0088] Furthermore, in order to disperse multiple samples 4B within the gel sheet 4 so that they are spaced apart from each other, for example as shown in Figure 7, it is preferable that the gel solution be treated to ensure that the low-concentration sample 4B is sufficiently dispersed. Here, low concentration refers to, for example, 3.5 × 10⁻⁶ for a sample with a diameter of 10 μm. 4 This refers to a concentration of less than or equal to 1 / mL.

[0089] In the above, sample 4B is described as either a single (isolated) cell or an aggregate of multiple cells. However, sample 4B may be any microorganism other than a cell or a plant. Furthermore, sample 4B may be any non-biological material selected from the group consisting of metals, plastics, proteins, oils, and polysaccharides.

[0090] In the gel sheet chip 6 described above, the gel 4D may be made of a material that has adhesive properties to cells. This allows the sample 4B to be adhered to and supported on the surface of the gel 4D. Even when the gel 4D embeds the sample 4B, if the gel 4D has adhesive properties, the sample 4B will be supported by the gel 4D more reliably.

[0091] Gel 4D is preferably made of a material having properties that allow for decomposition and dissolution. Figure 33 is a schematic cross-sectional view showing a first example of a process in which the gel is dissolved from the gel sheet portion and only the cells are recovered. Referring to Figure 33, it is also possible to recover only the cells (sample 4B) by dissolving the gel 4D that supports and fixes sample 4B from the cut-out gel sheet portion 4E. Specifically, for example, sample 4B may be 5 × 10 5 A gel sheet portion 4E, in which cells / mL of fibroblasts are placed inside gel 4D, is housed inside the apparatus. A gel dissolving agent 10, such as a 1% EDTA (ethylenediaminetetraacetic acid) solution, is supplied inside the apparatus. In Figure 33, gel 4D consists of a mixture of alginic acid with a content of 1.0% and collagen with a content density of 1.2 mg / mL. The gel dissolving agent 10 dissolves gel 4D. As a result, as shown in the lower part of Figure 33, the gel dissolving agent 10 contains the dissolved gel 4D and accumulates in the substrate as gel dissolving agent 10A. Since this is liquid, only the sample 4B can be recovered by discarding it.

[0092] Figure 34 is a schematic cross-sectional view showing a second example of the process in which the gel is dissolved from the gel sheet portion and only the cells are recovered. Referring to Figure 34, the sample 4B contained in the gel sheet portion 4E may be adhered to the outside of the gel 4D. In this case as well, similar to Figure 33, a gel dissolving agent 10, for example, a 1% EDTA solution, is supplied inside the apparatus.

[0093] Gel 4D is preferably made of a material that allows for the decomposition of crosslinking points and gel material molecules. This allows for the decomposition of gel 4D from gel sheet portion 4E after it has been cut off, thereby extracting only sample 4B. For example, consider the case where sodium alginate is used as gel 4D. In this case, by adding alginate lyase or a chelating agent (such as EDTA), the alginate gel can be dissolved to obtain sample 4B. Alginate lyase is an alginate-degrading enzyme. Chelating agents remove calcium ions necessary for crosslinking the alginate gel.

[0094] Alginate lyase has high specificity for alginate. EDTA primarily chelates calcium and magnesium ions in the culture medium. Therefore, methods using these enzymes have low toxicity to cells and are less invasive in sample collection.

[0095] This embodiment discloses a method for manufacturing a gel sheet chip 6 on which a sample 4B to be collected is fixed. In this method for manufacturing the gel sheet chip 6, a frame 3 with through-holes 3A formed therein is placed on a film 2. A fluid sample 4C containing the sample 4B to be collected is supplied into the through-holes 3A on the film 2. The thickness of the fluid sample 4C is made uniform by sandwiching the structure, which consists of the film 2, frame 3, and fluid sample 4C, between flat plates (slide glass 1, 5) placed on the upper and lower sides. The fluid sample 4C is then gelled. As a result, the thickness of the gel raw material 4A is controlled, and a gel sheet chip 6 can be provided that allows for stable, reproducible, and highly accurate collection of the sample 4B without damaging it.

[0096] (modified version) The embodiment described above may have the following features. The features of each of the following components may be combined as appropriate.

[0097] Firstly, from the viewpoint of suppressing the problem of the shape of the gel sheet portion 4E, which is housed by the pin 7B upon puncture, collapsing, for example, in the hollow portion 7C, the inner wall surface of the pin 7B may have a tapered shape extending in a direction inclined with respect to the direction in which the pin 7B extends, for example, at its tip. The angle of the tapered shape with respect to the direction in which the pin 7B extends may be, for example, 90° or less.

[0098] Secondly, the needle used in this embodiment is not limited to pin 7B, but can be any needle having a hollow section. For example, it may be an injection needle in which the hollow section is cylindrical (i.e., the entire needle is cylindrical).

[0099] Thirdly, from the viewpoint of easily discharging the stored gel sheet portion 4E, it is preferable that the hollow-shaped portion 7C used in this embodiment is coated with a material that has low protein adsorption properties and low affinity with gel 4D. Specifically, the inner wall surface of the hollow-shaped portion 7C may be coated with, for example, a fluorine-based material. Here, gel 4D is, for example, MPC polymer (2-methacryloyloxyethyl phosphorylcholine).

[0100] Furthermore, in this embodiment, gel 4D is not limited to commonly used materials such as the collagen gel described above, but may be formed from any of the following selected materials: tamarind seed gum, pectin, and carrageenan.

[0101] The features of each example described in the embodiments above may be applied in appropriate combinations within a technically consistent range.

[0102] The embodiments disclosed herein should be considered in all respects to be illustrative and not restrictive. The scope of the present invention is indicated by the claims rather than by the foregoing description, and all modifications within the meaning and scope equivalent to the claims are intended to be included. [Explanation of Symbols]

[0103] 1,5,5A Glass slides, 2 Film, 3 Frame, 3A,3C Through-holes, 3B Other frames, 4 Gel sheets, 4A Gel raw materials, 4B Sample, 4C Fluid sample, 4D Gel, 4E Gel sheet portion, 6,6A,6B Gel sheet tips, 7A Syringe, 7B Pin, 7C Hollow section, 7CP Syringe cap, 7D Upper syringe fixing section, 7E Lower syringe fixing section, 7F Holder shell, 7G Tube, 7H Collection needle holder, 9A Container, 10,10A Gel dissolving agent, 11 Sample collection set, 20 Culture medium, 40 First drive unit, 41 Servo motor, 43 Cam, 44 Bearing, 45 Cam connecting plate, 46 Movable part, 100 XY stage for container, 101 XY stage for sample, 101A Stage penetration section, 102 XY stage for container, 104 sampling mechanism, 106 observation optical system.

Claims

1. A gel sheet chip on which the sample to be collected is fixed, Film and, A frame placed on the aforementioned film and having through holes formed therein, A gel placed on the film within the through hole, The gel is supported by the sample to be collected, and the collection is comprised of the sample to be collected. The sample is placed on the upper surface of the gel so as to adhere to the gel. A gel sheet chip further comprising another frame placed on the frame so as to surround the sample on the upper surface of the gel.

2. The gel sheet chip according to claim 1, wherein the thickness of the gel is less than or equal to the thickness of the frame.

3. The gel sheet chip according to claim 1 or 2, wherein the sample is either cells or cell aggregates.

4. The gel sheet chip according to any one of claims 1 to 3, wherein the gel is made of a material that has adhesive properties to cells.

5. The gel sheet chip according to any one of claims 1 to 4, wherein the gel is a gel made from a biocompatible gel raw material.

6. The gel sheet chip according to any one of claims 1 to 5, wherein the gel is made of a material having properties that allow for decomposition and dissolution.

7. A method for manufacturing a gel sheet chip on which a sample to be collected is fixed, A step of installing a frame with through holes formed on the film, The steps include supplying a fluid sample containing the sample to be collected into the through-hole on the film, A step of making the thickness of the fluid sample uniform by sandwiching the structure, which consists of the film, the frame, and the fluid sample, between flat plates placed on the upper and lower sides of the structure, A method for manufacturing a gel sheet chip, comprising the step of gelling the aforementioned fluid sample.

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