Container for embedding, embedding method and embedding device

Abstract

The invention provides a container for embedding. An embedding container according to one embodiment of the present invention is used for accommodating an object to be embedded and is cut together with the object, and a surface near the object at a placement site where the object is placed has a cross-section different from a surface adjacent to the surface near the surface. Also provided is an embedding method including a placement step for placing an object in the embedding container and an embedding step for embedding the object together with the embedding container. Also provided is an embedding device comprising a moving unit that moves the embedding container and an introduction unit that introduces a treatment liquid and / or an embedding medium into the embedding container.

Description

Technical Field

[0001] The present invention relates to an embedding container, an embedding method using the embedding container, and an embedding device. Background Art

[0002] In cell biology and medical research, cells and tissues are fixed with formalin and embedded in paraffin to prepare specimens. For example, Patent Document 1 discloses a "tissue cassette set for embedding cell blocks." There is a need for more methods for embedding cells and tissues.

[0003] Prior art literature

[0004] Patent Literature

[0005] Patent Document 1: Japanese Patent No. 5204810 Summary of the Invention

[0006] An embedding container is provided. The embedding container can accommodate an object to be embedded. The embedding container can be cut together with the object. Any of the above embedding containers can include a retaining member that holds the object and allows passage of an embedding agent for embedding the object. Any of the above retaining members can be embedded in the embedding agent together with the object. Any of the above retaining members can be embedded while in contact with the object. A surface of any of the above embedding containers, where the object is to be placed, near the object can have a cross-section different from a surface adjacent to the near surface.

[0007] Any of the above-described embedding containers may have a first cross-section that is a cut surface of the embedding container and includes the proximal surface. Any of the above-described embedding containers may have a second cross-section that is a cut surface of the embedding container and includes a surface adjacent to the proximal surface and has a shape different from the shape of the first cross-section.

[0008] Any of the above-described embedding containers may have a first cross-section as a cut surface of the embedding container that includes the proximal surface. Any of the above-described embedding containers may have a third cross-section that is a cut surface of the embedding container that includes a surface adjacent to the proximal surface and has a cross-sectional area different from that of the first cross-section.

[0009] Any of the above-mentioned embedding containers may have a first cross-section as a cut surface of the embedding container including the proximal surface. Any of the above-mentioned embedding containers may have a fourth cross-section, the fourth cross-section being a cut surface of the embedding container including a surface adjacent to the proximal surface, the fourth cross-section being made of a material of the embedding container different from the material of the embedding container in the first cross-section.

[0010] The vicinity surface may be parallel to a placement surface on which any of the above-mentioned embedding containers is placed.

[0011] Any of the above-mentioned embedding containers may include an upper portion defining an upper space and a lower portion defining a lower space. The proximal surface may be arranged at a boundary between the upper portion and the lower portion.

[0012] The upper portion may have an open shape opened to the outside.

[0013] The lower portion may have a hole that can be connected to the outside of any of the above-mentioned embedding containers.

[0014] Any of the above-mentioned embedding containers may include a first porous member on the proximal surface. The first porous member may be the holding member.

[0015] Any of the above-mentioned embedding containers may further include a second porous member at a position facing the first porous member with the object interposed therebetween. The second porous member may be the holding member.

[0016] The thickness of the first porous member in a direction perpendicular to the installation surface on which any of the above-mentioned embedding containers is installed may be 100 μm or less.

[0017] Any of the above-mentioned embedding containers may include an upper portion defining an upper space and a lower portion defining a lower space with the retaining member as a reference. The cross-sectional area of ​​the lower portion may differ from the cross-sectional area of ​​the upper portion. The circumferential length of the lower portion may differ from the circumferential length of the upper portion. Any of the above-mentioned embedding containers may have a step formed at the boundary between the upper portion and the lower portion. Any of the above-mentioned first porous members may be disposed on the step.

[0018] Any of the above-mentioned embedding containers may include a water absorbing member disposed in the lower space. The water absorbing member may be the holding member.

[0019] The water absorbing member may have a plurality of colors. In any of the above-mentioned embedding containers, the color of the water absorbing member may be switched at the proximal surface.

[0020] The lower space may have an inner diameter of the embedding container that gradually increases as approaching the upper portion in a direction perpendicular to a mounting surface on which any of the above-mentioned embedding containers is mounted.

[0021] The upper space may have an inner diameter of the embedding container that gradually decreases as it approaches the lower portion in a direction perpendicular to a mounting surface on which any of the above-mentioned embedding containers is mounted.

[0022] The height of the upper portion may be greater than 3 mm and less than 20 mm. The height of the lower portion may be greater than 0.2 mm and less than 3 mm. The inner diameter of any of the above embedding containers may be greater than 0.5 mm and less than 6 mm.

[0023] The plurality of upper spaces of any of the above-mentioned embedding containers may be arranged in a planar shape.

[0024] Any of the above-mentioned embedding containers may form a honeycomb structure by the upper spaces arranged in the planar shape.

[0025] Any of the above-mentioned embedding containers may be formed of at least one of plastic, paper, wood, and fiber.

[0026] The object may include at least one of cells, cell aggregates, biological tissues, organisms, fungi, and algae.

[0027] Any of the above embedding containers may further include an outer peripheral portion surrounding the outer periphery of the embedding container. The outer peripheral portion may have a surface that is not perpendicular to a direction in which any of the above embedding containers is cut.

[0028] The outer peripheral portion may include an intermediate portion extending in the radial direction of the embedding container between the outer peripheral portion and any of the above embedding containers. The intermediate portion may be a region extending 3 mm or more relative to the direction in which the embedding container is cut.

[0029] An embedding method using any of the above embedding containers is provided. The embedding method may include a placement step of placing the object in any of the above embedding containers and an embedding step of embedding the object together with any of the above embedding containers.

[0030] Any of the above embedding methods may include a pretreatment step after the placement step and before the embedding step, wherein a treatment liquid is introduced into any of the above embedding containers with the object placed therein to pretreat the object.

[0031] Any of the above embedding methods may include an embedding step in a container after the pretreatment step and before the embedding step, wherein, with the object disposed in any of the above embedding containers, an embedding agent is introduced into the embedding container to embed the object in the embedding container.

[0032] An embedding device having any of the above embedding containers is provided. The embedding device may include a moving portion for moving any of the above embedding containers and an introduction portion for introducing a treatment liquid and / or an embedding agent into the embedding container. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 An example of the embedding container in this embodiment is shown.

[0034] Figure 2 An example of the embedding container in this embodiment is shown.

[0035] Figure 3 An example of the embedding container in this embodiment is shown.

[0036] Figure 4 An example of the embedding container in this embodiment is shown.

[0037] Figure 5 An example of the embedding container in this embodiment is shown.

[0038] Figure 6 An example of the embedding container in this embodiment is shown.

[0039] Figure 7 An example of the embedding container in this embodiment is shown.

[0040] Figure 8 An example of the embedding container in this embodiment is shown.

[0041] Figure 9 An example of the embedding container in this embodiment is shown.

[0042] Figure 10 An example of the embedding container in this embodiment is shown.

[0043] Figure 11 An example of the embedding container in this embodiment is shown.

[0044] Figure 12 An example of the aggregation structure of embedding containers in this embodiment is shown.

[0045] Figure 13 An example of the aggregation structure of embedding containers in this embodiment is shown.

[0046] Figure 14 An example of the embedding container in this embodiment is shown.

[0047] Figure 15 An example of the configuration of the embedding device in this embodiment is shown.

[0048] Figure 16 An example of the flow of a method for sampling an object in this embodiment is shown.

[0049] Figure 17 An example of the flow of a method for placing an object in an embedding container in this embodiment is shown.

[0050] Figure 18 This figure explains the method of placing the object in the embedding container at S100 in this embodiment.

[0051] Figure 19 This figure explains the method of placing the object in the embedding container at S100 in this embodiment.

[0052] Figure 20 An example of a flow of a method for pre-processing an object in this embodiment is shown.

[0053] Figure 21 A diagram showing a method of pre-processing an object in this embodiment.

[0054] Figure 22 An example of an embedding container filled with an embedding medium in this embodiment is shown.

[0055] Figure 23 An example of an embedding container filled with an embedding medium in this embodiment is shown.

[0056] Figure 24 This is a diagram illustrating a method of slicing an object in this embodiment.

[0057] Figure 25 This is a diagram illustrating an example of sampling of the clustered structure of the embedding container in the present embodiment.

[0058] Figure 26 It is a diagram for explaining S170 and S190 in an example of sampling of the clustered structure of the embedding container.

[0059] Figure 27 This is a diagram illustrating an example of sampling of the clustered structure of the embedding container in the present embodiment.

[0060] Figure 28 An example of a cross-sectional view of an embedding container included in the aggregation structure is shown.

[0061] Figure 29 It is a diagram illustrating steps S100 to S270 in an example of specimenization of the clustered structure of the embedding container.

[0062] Figure 30 This is a diagram illustrating S190 in an example of sampling of the clustered structure of the embedding container.

[0063] Figure 31 An example of the hardware configuration of a computer is shown. DETAILED DESCRIPTION

[0064] The invention is described below using embodiments thereof. However, the embodiments do not limit the invention as defined in the claims. Furthermore, not all combinations of features described in the embodiments are essential to the invention's solution. It should be noted that in the drawings, identical or similar parts may be denoted by the same reference numerals, and duplicate descriptions may be omitted.

[0065] Figure 1 An example of the configuration of an embedding container 100 in this embodiment is shown. The embedding container 100 can accommodate and arrange an object 115 therein. By embedding the embedding container 100 containing the object 115 and then cutting the embedding container 100 and the object 115 together, a specimen of the object 115 can be prepared. Details will be described later.

[0066] The embedding container 100 can be placed on a setting surface 180. For example, the setting surface 180 is a horizontal surface. The horizontal surface here is not limited to a horizontal surface, and can be inclined by ±10 degrees or less, ±9 degrees or less, ±8 degrees or less, ±7 degrees or less, ±6 degrees or less, ±5 degrees or less, ±4 degrees or less, ±3 degrees or less, ±2 degrees or less, or ±1 degree or less relative to the horizontal surface. Figure 1 This is a cross-sectional view of the embedding container 100 when cut along a cross section perpendicular to the installation surface 180 .

[0067] When the embedding container 100 is cut in any plane, a cross section of the wall surface 175 serving as the wall of the embedding container 100 can be revealed on the cut plane. The wall surface 175 may be a side surface of the embedding container 100 or a bottom surface.

[0068] The embedding container 100 may include all or at least a portion of a container body 110 and a retaining member. The retaining member may be a porous member. As an example, the porous member is a filter 118. As an example, the porous member is a water-absorbing member. Details of the retaining member will be described later.

[0069] The container body 110 constitutes the main structure of the embedding container 100. The container body 110 may be in the shape of a hollow cylinder (column), a hollow polygonal prism, a hollow truncated cone, or a hollow polygonal pyramid, but is not limited thereto.

[0070] The container body 110 can be made of at least one of plastic, paper, wood, and fiber. The container body 110 can be made of at least one of plastic, paper, wood, and fiber, making it easier to cut the embedding container 100 when preparing a specimen containing the object 115.

[0071] The plastic may be a thermoplastic resin or a thermosetting resin. For example, the plastic includes one or more of silicone rubber, polyacetal (POM), polycarbonate (PC), polypropylene (PP), polyethylene terephthalate (PET), polyurethane (PU), polymethyl methacrylate (PMMA), polystyrene (PS), and polyvinyl chloride (PVC), but is not limited thereto.

[0072] When the container body 110 is made of plastic, it can be manufactured by any method such as compression molding, injection molding, or extrusion molding. Plastic has excellent processability. When the container body 110 is made of wood, it can be manufactured by any method such as thermoforming. When the container body 110 is made of paper or fiber, it can be manufactured by any method such as bonding.

[0073] Object 115 is an object for specimen preparation. Object 115 is a living organism. The living organism may be an organic life form. For example, the living organism is a cell. As an example, the cell is an animal cell or a plant cell. As an example, the cell is a living cell or a dead cell.

[0074] The organism may also be an organism other than a cell. As an example, the organism includes at least one of a single cell, a sphere (cell aggregate), a biological tissue, an organoid (cell aggregate), an organism (microorganism, etc.), a fungus, and an algae. The organism may also include organelles within the cell. The size of the organism may be a long diameter of less than 0.5 mm, less than 1 mm, less than 1.5 mm, less than 2 mm, less than 2.5 mm, or less than 3 mm, but is not limited thereto. The size of the organism may be a long diameter of less than 100 μm or less than 500 μm (the size of a microstructure) that is not easily visually confirmed.

[0075] The interior of the embedding container 100 and the object 115 are immersed in a liquid 160. The liquid 160 can be a culture medium for culturing the object 115 and / or a liquid for preparing a specimen. For example, the liquid 160 can be water, a buffer solution, a culture medium, or an organic solvent. The organic solvent can be a treatment solution or embedding solution described later, or a fixative, dehydrating solution, defatting solution, and / or embedding medium used for preparing a specimen. As an example, the organic solvent is formalin (formaldehyde, paraformaldehyde), glutaraldehyde, Bouin's solution, Carnoy's solution, methanol, ethanol, xylene, a xylene substitute, paraffin, and / or collodion.

[0076] The retaining member retains the object 115 thereon. The retaining member can be embedded together with the object 115 while retaining the object 115. As an example, the retaining member is embedded in a state of contact with the object 115. For example, the retaining member can be a porous member. The porous member can be a member having holes on the surface and / or inside. For example, the porous member is a filter 118. The filter 118 allows the liquid 160 to pass through and retains the object 115. That is, the object 115 is arranged on the filter 118. The filter 118 can be fixed to the container body 110. The filter 118 can be fixed to the container body 110 by welding using a laser or by bonding using double-sided tape or an adhesive. By laser welding the filter 118 to the container body 110, the container body 110 can withstand the pressure associated with the introduction and discharge of the liquid 160, and the container body 110 is not corroded by the liquid 160. When double-sided tape or an adhesive is used for bonding, the double-sided tape or adhesive is preferably made of a material that does not dissolve due to the heated liquid 160, and does not fall off and / or deteriorate due to the introduction and discharge of the liquid 160. During the step of pre-treating the object 115 (e.g., step S200 described later), the filter 118 can be removed from the container body 110 as needed.

[0077] A gap may be provided between the filter 118 and the wall surface 175 of the container body 110. The liquid 160 can pass through the gap. The major or minor diameter of the gap in the filter 118 through which the liquid 160 can pass can be larger than the major or minor diameter of the object 115. Since the major or minor diameter of the gap through which the liquid 160 can pass is larger than the major or minor diameter of the object 115, the object 115 does not obstruct the passage of the liquid 160.

[0078] Instead of or in addition to providing gaps, filter 118 may include one or more holes (perforations) to allow liquid 160 to flow between upper portion 120 and lower portion 125. The size of the holes can be arbitrarily selected so that object 115 does not pass through. The hole size can be approximately 0.2 μm or larger in diameter and approximately 10 μm or smaller in diameter, but is not limited thereto.

[0079] The material of the filter 118 is a material that is not corroded by the liquid 160 and / or a material that can withstand the pressure associated with the introduction and discharge of the liquid 160, and can be used without particular restrictions. The filter 118 can be a mesh filter. For example, the material of the filter 118 is at least one of plastic, paper, wood, and fiber. As an example, the material of the filter 118 is a plastic such as polytetrafluoroethylene (PFTE), polyimide (PI), or polyethersulfone (PES). The filter 118 can also be a non-woven fabric. For example, the material of the filter 118 is fiber.

[0080] The surface of the filter 118 that contacts the object 115 may be coated to have affinity with the object 115. Examples of the coating material include, but are not limited to, poly-L-lysine, poly-D-lysine, extracellular matrix (laminin, fibronectin, collagen, etc.), and proteoglycan.

[0081] The thickness of the filter 118 in a direction perpendicular to the installation surface 180 on which the embedding container 100 is installed can be 20 μm or greater and 200 μm or less. The thickness of the filter 118 in a direction perpendicular to the installation surface 180 on which the embedding container 100 is installed can be 100 μm or less. When the thickness of the filter 118 is 100 μm or greater and 200 μm or less, the embedding container with the filter 118 can be manufactured and handled easily. When the thickness of the filter 118 is 20 μm or greater and less than 100 μm, the pressure required to communicate the liquid 160 can be reduced, which is advantageous for handling the embedding solution.

[0082] The embedding container 100 can be divided into an upper portion 120 and a lower portion 125. For example, the embedding container 100 is divided into the upper portion 120 and the lower portion 125 with the position of the filter 118 as the boundary. Thus, the filter 118 is provided at the boundary between the upper portion 120 and the lower portion 125. The filter 118 can serve as a placement location for placing the object 115.

[0083] The upper portion 120 defines the upper space of the embedding container 100. The upper space can be defined based on the retaining member. In this case, the space above the retaining member is the upper space. All or part of the upper space can be filled with liquid 160. The upper portion can be open to the outside. For example, the upper space may not be covered. For example, to prevent dust and the like from entering the upper space, a cover can be provided on the open embedding container 100 to seal the upper space. To place the object 115 in the upper space, a nozzle of the biological manipulation unit, described later, can be inserted. Details will be provided later.

[0084] The lower portion 125 defines the lower space of the embedding container 100. The lower space can be defined relative to the retaining member. In this case, the space below the retaining member, i.e., the space opposite the upper space relative to the retaining member, constitutes the lower space. All or part of the lower space can be filled with liquid 160. The lower space can be externally sealed.

[0085] The lower portion 125 of the container body 110 may be provided with a hole 130 that allows connection to the exterior of the embedding container 100. Liquid 160 can be introduced from the exterior of the embedding container 100 into the lower portion 125 and upper portion 120 through the hole 130. To embed or pre-treat the object 115, embedding or pre-treating can be performed using a small amount of liquid 160 while filling a portion of the upper and lower spaces with liquid 160. Liquid 160 can be discharged from the lower portion 125 to the exterior of the embedding container 100 through the hole 130. By filling the lower space with liquid 160 and allowing the liquid to flow through the filter 118 toward the upper space, the upper space can be filled with liquid 160. The hole 130 can be formed on the bottom or side of the lower portion 125. The diameter of the hole 130 can be arbitrarily determined according to the size of the object 115. If the diameter of hole 130 is larger than the diameter of object 115, hole 130 will not be blocked by object 115, which can solve the problem of hole 130 being blocked by object 115. Hole 130 can be blocked by a cap, plug, etc. except when liquid 160 is introduced into container body 110 and / or discharged from container body 110.

[0086] Here, the upper portion 120 and the lower portion 125 may be defined as the portion located relatively above in the height direction of the embedding container 100 (a direction perpendicular to the installation surface 180) as the upper portion 120, and the portion located relatively below as the lower portion 125. When the embedding container 100 is placed on the installation surface 180, the portion relatively far from the installation surface 180 may be defined as the upper portion 120, and the portion relatively close to the installation surface may be defined as the lower portion 125.

[0087] The liquid 160 may be directly introduced into the upper space without passing through the hole 130. The upper portion 120 and the lower portion 125 may be fixed integrally or may be separable as needed.

[0088] Nearby surface 170 is a surface located near the placement location for object 115. Nearby surface 170 may be located at the boundary between upper portion 120 and lower portion 125. Nearby surface 170 may include the surface on which object 115 is placed. Nearby surface 170 may include the surface on which object 115 is placed and a surface located near the surface on which object 115 is placed.

[0089] The nearby surface 170 may include the surface on which the object 115 is placed and surfaces existing in a space within a predetermined height range in two directions relative to the height direction (perpendicular to the installation surface 180) of the surface on which the object 115 is placed. For example, the predetermined height is approximately 10 times, approximately 9 times, approximately 8 times, approximately 7 times, approximately 6 times, approximately 5 times, approximately 4 times, approximately 3 times, approximately 2 times, or approximately 1 time the major or minor axis of the object 115.

[0090] The proximal surface 170 may be parallel to the installation surface 180 on which the embedding container 100 is installed. For example, the proximal surface 170 is a horizontal surface. The proximal surface 170 is not limited to a horizontal surface and may be inclined relative to the horizontal surface by ±10 degrees or less, ±9 degrees or less, ±8 degrees or less, ±7 degrees or less, ±6 degrees or less, ±5 degrees or less, ±4 degrees or less, ±3 degrees or less, ±2 degrees or less, or ±1 degree or less.

[0091] The nearby surface 170 may be perpendicular to the installation surface 180 on which the object 115 is placed. In this case, the nearby surface 170 may include a surface located inside a cylinder on the installation surface 180 with reference to the location on which the object 115 is placed, wherein the cylinder has a side surface enclosed by a circle with a radius of about 10 times, about 9 times, about 8 times, about 7 times, about 6 times, about 5 times, about 4 times, about 3 times, about 2 times, or about 1 times the length of the major or minor axis of the object 115.

[0092] The vicinity surface 170 may also include a porous member. The vicinity surface 170 may also be a surface included in a porous member (e.g., filter 118) on which the object 115 is placed. In this case, the vicinity surface 170 may also be a surface having a shape that matches the shape of the porous member.

[0093] A surface adjacent to the nearby surface 170 can be defined as a surface adjacent to the nearby surface 170. The nearby surface 170 and the surface adjacent to the nearby surface 170 may be connected at a point, or may be connected by a straight line, a curve, a plane, or a curved surface. As an example, a surface adjacent to the nearby surface 170 is a surface parallel to the nearby surface 170 and is within a height range included in either the height x of the upper portion 120 or the height y of the lower portion.

[0094] When the liquid 160 is introduced and / or discharged, the near surface 170 of the embedding container 100 may be inclined by ± about 10 degrees or less, ± about 9 degrees or less, ± about 8 degrees or less, ± about 7 degrees or less, ± about 6 degrees or less, ± about 5 degrees or less, ± about 4 degrees or less, ± about 3 degrees or less, ± about 2 degrees or less, or ± about 1 degree or less relative to the installation surface 180 and the horizontal plane.

[0095] The adjacent surface 170 of the object 115 and the adjacent surface 170 may have different cross-sections. For example, the adjacent surface 170 and the adjacent surface 170 may differ in one or more of their cross-sectional shape, cross-sectional area, and cross-sectional material. The cross-sectional area may or may not include the object 115. By using one or more of the cross-sectional shape, cross-sectional area, and cross-sectional material as a criterion, the embedding container 100 of this embodiment allows for easy preparation of a specimen of the object 115 without losing the object 115. This will be described in detail below.

[0096] Specifically, the embedding container 100 may have a first cross section, which is a cut surface of the embedding container 100 including the proximal surface 170, and a second cross section, which is a cut surface of the embedding container 100 including a surface adjacent to the proximal surface 170 and has a shape different from the shape of the first cross section. The shapes of the first and second cross sections may be, but are not limited to, a circle, an ellipse, or a polygon (such as a triangle, a quadrilateral, a pentagon, or a hexagon). Details will be described later.

[0097] The embedding container 100 may have a first cross section, which is a cut surface of the embedding container 100 including the proximal surface 170, and a third cross section, which is a cut surface of the embedding container 100 including a surface adjacent to the proximal surface 170 and has a cross-sectional area different from that of the first cross section. The cross-sectional areas of the first and third cross sections may be the areas of the cross section enclosed by the outer diameter of the embedding container 100 or the areas of the cross section enclosed by the inner diameter of the embedding container 100. The difference in cross-sectional area between the first and third cross sections allows for the formation of a step difference in the embedding container 100, as will be described in detail later.

[0098] The embedding container 100 may have a first cross section, which is a cut surface of the embedding container 100 including the proximal surface 170, and a fourth cross section, which is a cut surface of the embedding container 100 including a surface adjacent to the proximal surface 170 and has a material of the embedding container 100 different from the material of the embedding container 100 in the first cross section. The material of the embedding container 100 in the first and fourth cross sections may be at least one of plastic, paper, wood, and fiber.

[0099] The height x (the length in the direction perpendicular to the bottom surface) of the upper portion 120 of the embedding container 100 can be 3 mm or more and 5 mm or less. When the height x of the upper portion 120 is within this range, the time required for temperature regulation of the liquid 160 and energy consumption are improved. The height x (the length in the direction perpendicular to the bottom surface) of the upper portion 120 of the embedding container 100 can be 5 mm or more and 20 mm or less. When the height x of the upper portion 120 is within this range, the embedding container 100 can be easily mounted on a conventional microtome.

[0100] The height y (the length perpendicular to the bottom surface) of the lower portion 125 of the embedding container 100 can be 0.2 mm or greater and 3 mm or less. By ensuring that the height y of the lower portion 125 is within this range, the time required for temperature regulation of the liquid 160 and energy consumption are improved, and the number of cuts is reduced. Furthermore, in the embedding method described below, it is advantageous to house the embedding container 100 in an embedding dish.

[0101] The inner diameter z of the upper portion 120 and / or the lower portion 125 of the embedding container 100 in a direction horizontal to the bottom surface may be 0.5 mm or greater and 6 mm or less. When the inner diameter z of the embedding container 100 is within the range of 0.5 mm or greater and 6 mm or less, the embedding container 100 can be appropriately accommodated in the embedding dish, and operability is also excellent.

[0102] The embedding container 100, in which the object 115 is placed on a porous member (e.g., filter 118), can be embedded in paraffin wax or the like and then cut together with the embedding container 100 to obtain a specimen of the object 115. The embedding container 100 can be cut parallel to the bottom surface of the lower portion 125 of the embedding container 100, from the bottom surface toward the upper portion 120. Instead of cutting parallel to the bottom surface, the embedding container 100 can be cut parallel to a plane that tilts the embedding container 100 from the horizontal by ±10 degrees or less, ±9 degrees or less, ±8 degrees or less, ±7 degrees or less, ±6 degrees or less, ±5 degrees or less, ±4 degrees or less, ±3 degrees or less, ±2 degrees or less, or ±1 degree or less.

[0103] When cutting the embedding container 100, rough cutting can be performed before the porous member appears. In this case, the cross section where the porous member appears can become the cross section of the adjacent surface 170, and the cross section before the porous member appears can become the cross section of the surface adjacent to the adjacent surface 170. The presence of the porous member on the cross section of the adjacent surface 170 changes one or more of its cross-sectional shape, cross-sectional area, and material. Since these changes indicate the presence of the object 115 in its vicinity, the adjacent surface 170 can be cut carefully.

[0104] By adjusting the cutting of the embedding container 100 based on the presence of the porous member, a specimen of the object 115 can be easily prepared without losing the object 115. Details will be described later.

[0105] Figure 2 Another example of the structure of the embedding container 100 in this embodiment is shown. Figure 1 In the embedding container 100 shown in FIG. 1 , the angle formed between the bottom surface and the side surface of the embedding container 100 is substantially perpendicular. Figure 2 In the embedding container 100 a shown, the side surfaces of the embedding container 100 are not perpendicular to the bottom surface. Figure 2 The embedding container 100a shown has an inner diameter that increases as it approaches the upper portion 120 in a direction perpendicular to the installation surface 180 (height direction), but the present invention is not limited thereto. The inner diameter may also decrease as it approaches the upper portion 120 in a direction perpendicular to the installation surface 180 (height direction). Figure 2The illustrated embedding container 100a has an inner diameter that increases continuously as it approaches the upper portion 120 in a direction perpendicular to the installation surface 180 (height direction), but the invention is not limited thereto and the inner diameter may increase / decrease intermittently.

[0106] Figure 2 When the embedding container 100a shown is cut horizontally from the bottom surface of the lower portion 125 toward the upper portion 120 of the embedding container 100a, the cross-sectional area of ​​the cut surface gradually increases. Therefore, when a predetermined cross-sectional area value (e.g., the cross-sectional area value near the boundary between the upper portion 120 and the lower portion 125) is reached, it indicates that the object 115 is located nearby. Therefore, when cutting the embedding container 100a, a rough cut can be performed before the predetermined cross-sectional area value is reached, and after the predetermined cross-sectional area value is reached, the nearby surface 170 can be carefully cut. In this way, the embedding container 100a can be cut in an adjustable manner, targeting the predetermined cross-sectional area value, making it possible to easily prepare a specimen of the object 115 without losing the object 115.

[0107] Figure 3 Another example of the structure of the embedding container 100 in this embodiment is shown. Figure 1 In the embedding container 100 shown, the cross-sectional area of ​​the upper portion 120 and the cross-sectional area of ​​the lower portion 125 of the embedding container horizontal to the bottom surface are substantially equal. Figure 3 The cross-sectional area and / or circumferential length (e.g., the length of the outer circumference of the cross section parallel to the bottom surface of the embedding container 100b) of the lower portion 125 of the embedding container 100b shown in FIG. are different from the cross-sectional area and / or circumferential length (e.g., the length of the outer circumference of the cross section parallel to the bottom surface of the embedding container 100b) of the upper portion 120. As a result, a step 173 is formed at the boundary between the upper portion 120 and the lower portion 125. The filter 118, which is a porous member, can be placed above the step 173.

[0108] Figure 3 The illustrated embedding container 100b has a smaller cross-sectional area at the bottom surface of the lower portion 125 than at the bottom surface of the upper portion 120. However, the embodiment is not limited thereto. The cross-sectional area at the bottom surface of the lower portion 125 may be larger than the cross-sectional area at the bottom surface of the upper portion 120. In this case, a step 173 is also formed at the boundary between the upper portion 120 and the lower portion 125, and the filter 118, which is a porous member, can be disposed above the step 173.

[0109] At once Figure 3In the embedding container 100b shown, when cutting from the bottom surface of the lower portion 125 toward the upper portion 120, parallel to the bottom surface, the embedding container 100b can be roughly cut until a step 173 appears on the cross section. The presence of a step 173 on the cut surface indicates the presence of the object 115 nearby, allowing for careful cutting. The reaching of the step 173 can be determined when the cross-sectional area occupied by the wall 175 on the cut surface increases or when the shape of the wall 175 on the cut surface changes. By adjusting the cutting of the embedding container 100b based on the presence of the step 173, specimens of the object 115 can be easily prepared without losing the object 115.

[0110] At once Figure 3 In the illustrated embedding container 100b, the inner diameter of the embedding container 100b in the lower space is substantially constant in a direction horizontal to the bottom surface. However, the inner diameter of the lower space may gradually increase or decrease as it approaches the upper portion 120 in a direction perpendicular to the installation surface 180 on which the embedding container 100b is installed (in the height direction). In this case, when the embedding container 100b is cut from the bottom surface of the lower portion 125 toward the upper portion 120, the cross-sectional area occupied by the wall surface 175 in the cut surface gradually increases or decreases. Therefore, when the cross-sectional area occupied by the wall surface 175 reaches a predetermined value (for example, the cross-sectional area occupied by the wall surface 175 near the boundary between the upper portion 120 and the lower portion 125), it becomes clear that the object 115 is present nearby. Therefore, the embedding container 100 b can be cut adjustably based on the predetermined value of the cross-sectional area occupied by the wall surface 175 , and a specimen of the object 115 can be easily prepared without losing the object 115 .

[0111] Figure 4 Another example of the configuration of the embedding container 100 in this embodiment is shown. Figure 1 The side surface of the embedding container 100 shown is substantially flat. Figure 4 The embedding container 100 c shown has one or more protrusions 172 formed on a proximal surface 170 , the protruding protrusions 172 projecting toward the outside of the embedding container 100 .

[0112] At once Figure 4 In the embedding container 100c shown, when cutting from the bottom surface of the lower portion 125 toward the upper portion 120, parallel to the bottom surface, the rough cut can be made until a protrusion 172 appears on the cross section. The appearance of protrusion 172 on the cut surface indicates the presence of the object 115 nearby, allowing for careful cutting. By adjusting the cutting of the embedding container 100c based on the presence of protrusion 172, specimens of the object 115 can be easily prepared without losing the object 115.

[0113] Figure 4 The embedding container 100c shown has a protrusion 172, but one or more depressions may be formed instead of the protrusion 172. In this case, by adjusting the cutting of the embedding container 100c based on the presence of the depression, a specimen of the object 115 can be easily prepared without losing the object 115.

[0114] Figure 5 Another example of the configuration of the embedding container 100 in this embodiment is shown. The embedding container 100 may further include a porous member at a position facing the porous member with the object 115 interposed therebetween. Figure 1 In the embedding container 100 shown, the filter 118 as a porous member is composed of a sheet provided at the boundary between the upper portion 120 and the lower portion 125. Figure 5 The embedding container 100d shown includes a filter 118b on the side opposite to the boundary with respect to the object 115, in addition to the filter 118a provided on the boundary.

[0115] The filter 118a and the filter 118b may have the same shape. The filter 118b may be provided separately on the upper portion of the embedding container 100d or integrally with the lid 178 of the embedding container 100d. In this case, the lid 178 may be provided with the filter 118b on the side facing the object 115.

[0116] Figure 5 The embedding container 100d shown can reduce the possibility of the object 115 being lost by sandwiching the object 115 between two filters, the filter 118a and the filter 118b. Figure 5 The embedding container 100d shown can reduce the consumption of the liquid 160 by placing two filters close to each other, thus being economical.

[0117] Figure 6 Another example of the configuration of the embedding container 100 in this embodiment is shown. Figure 3 The lower space of the lower portion 125 of the embedding container 100b shown in FIG. 1 is a void. Figure 6 The embedding container 100e shown has a water absorbing member 190 disposed as a porous member in the lower space of the lower portion 125. The water absorbing member 190 absorbs the liquid 160. The water absorbing member 190 may occupy all or at least a portion of the lower space.

[0118] Water-absorbing member 190 can be used without particular limitation, as long as it can absorb liquid 160, such as water. For example, water-absorbing member 190 can be a sponge, woven fabric, non-woven fabric, foam, porous polymer, or water-absorbing high molecular compound. For example, water-absorbing member 190 is a sponge made of polyester, nylon, or cotton. Water-absorbing member 190 can also be a multi-layered structure made of different materials or textures. For example, water-absorbing member 190 can be a structure composed of two layers of polymers made of different materials.

[0119] In the embedding container 100e, the object 115 can be placed directly on the water absorbing member 190. In this case, the filter 118 may not be provided inside the embedding container 100e. If the filter 118 is provided above the water absorbing member 190 inside the embedding container 100e, the object 115 can be placed above the filter 118.

[0120] Figure 6 The embedding container 100e shown can absorb unnecessary liquid 160 within the embedding container 100e using the water absorbing member 190, thereby eliminating the need for drainage and reducing environmental impact. To fully absorb the unnecessary liquid 160, the cross-sectional area of ​​the lower portion 125 of the embedding container 100e can be larger than the cross-sectional area of ​​the upper portion 120. This increases the space below the lower portion 125, allowing for the larger water absorbing member 190 to be accommodated therein, resulting in improved absorption of the liquid 160.

[0121] exist Figure 6 In the embedding container 100e shown, a recessed portion can be provided by embossing or the like in the portion of the water absorbing member 190 where the object 115 is to be placed. Providing the recessed portion in the water absorbing member 190 allows the object 115 to be reliably placed in the recessed portion of the water absorbing member 190 even during the introduction and discharge of the liquid 160.

[0122] The water absorbing member 190 can be a single color. Alternatively, the water absorbing member 190 can have multiple colors. The color of the water absorbing member 190 can be switched at the near surface 170. The color can be any color. Switching from light to dark, dark to light, warm to cool, or cool to warm, etc., can provide excellent visual recognition.

[0123] By switching the color of the water absorbing member 190 on the nearby surface 170, it is possible to detect the presence of the object 115 nearby. Therefore, when cutting the embedding container 100e from the bottom surface of the lower portion 125 toward the upper portion 120, the cutting can be adjusted to target the color switching of the water absorbing member 190, making it possible to easily prepare a specimen of the object 115 without losing the object 115.

[0124] Figure 7Another example of the configuration of the embedding container 100 in this embodiment is shown. Figure 1 The inner diameters of the upper portion 120 and the lower portion 125 of the embedding container 100 shown are substantially constant. Figure 7 The upper space of the embedding container 100 f shown in the figure has an inner diameter that gradually decreases as it approaches the lower portion 125 along a direction (height direction) perpendicular to the installation surface 180 on which the embedding container 100 f is installed.

[0125] Figure 7 The upper space of the embedding container 100f shown has an inner diameter that gradually decreases as it approaches the lower portion 125 along a direction perpendicular to the installation surface 180 (height direction) on which the embedding container 100f is installed. However, the present invention is not limited thereto, and the inner diameter of the upper space may gradually increase as it approaches the lower portion 125 along a direction perpendicular to the installation surface 180.

[0126] At once Figure 7 In the illustrated embedding container 100f, the inner diameter of the upper space gradually decreases as it approaches the lower portion 125 in a direction perpendicular to the installation surface 180. This reduces the area (e.g., the filter 118) where the object 115 is placed, minimizing the likelihood of the object 115 being lost. Furthermore, during the introduction and discharge of the liquid 160, even if the object 115 floats from the filter 118, the inclined wall 175 allows the object 115 to slide onto the filter 118, thereby allowing the object 115 to be collected there.

[0127] Figure 7 Since the embedding container 100f shown has a small lower space, when the embedding container 100f is cut from the bottom surface of the lower portion 125 toward the upper portion 120, the distance to the object 115 can be shortened, thereby reducing the number of cuts.

[0128] Figure 7 The embedding container 100f shown has a truncated cone-shaped cross section (mortar-shaped) when cut perpendicularly to the installation surface 180. However, the cross section is not limited to this and may also be U-shaped or V-shaped. In this case, the object 115 also slides onto the filter 118 and can be collected on the filter 118.

[0129] In a configuration in which the inner diameter of the upper space of the embedding container 100f gradually increases as it approaches the lower portion 125 in a direction perpendicular to the installation surface 180, the opening of the upper space can be reduced, and thus the object 115 can be arranged close to the central axis of the embedding container 100.

[0130] Figure 8Another example of the configuration of the embedding container 100 in this embodiment is shown. Figure 1 The lower portion 125 of the embedding container 100 shown in FIG. 1 has a large lower space. Figure 8 The embedding container 100g shown is Figure 1 Compared with the embedding container 100 shown, the lower space of the lower portion 125 is smaller, and the inner diameter of the lower space of the lower portion 125 is substantially the same as the diameter of the hole 130 .

[0131] Figure 8 The embedding container 100 g shown has a small lower space, so the consumption of the liquid 160 can be small, which is economical.

[0132] Figure 8 The embedding container 100g shown is provided so that the filter 118 covers substantially the entire surface of the boundary between the upper portion 120 and the lower portion 125. However, the filter 118 may be provided so as to cover only a portion of the boundary between the upper portion 120 and the lower portion 125. In this case, the filter 118 can be made smaller, which is economical.

[0133] Figure 9 Another example of the configuration of the embedding container 100 in this embodiment is shown. Figure 8 The embedding container 100g shown in the figure is provided with a filter 118 so as to cover the boundary between the upper portion 120 and the lower portion 125. Figure 9 The embedding container 100h shown is provided with a water absorbing member 190 at the boundary between the upper portion 120 and the lower portion 125 so as to close the lower space. Figure 9 In the embedding container 100h shown, the inner diameter of the lower space may be substantially the same as the inner diameter of the hole 130. In addition, the filter 118 may be provided instead of or in addition to the water absorbing member 190.

[0134] Figure 9 The embedding container 100h shown is Figure 7 Compared with the embedding container 100f of FIG. 1 , the porous member (eg, the water absorbing member 190 ) can be made smaller, which is economical. Figure 9 The embedding container 100h shown has a small lower space, so the consumption of the liquid 160 can be small, which is economical.

[0135] Figure 8 The embedding container 100g shown has only one wall surface 175 constituting the lower portion 125. Figure 9 The embedding container 100h shown has two wall surfaces 175 constituting the lower portion 125.

[0136] Figure 9The embedding container 100h shown has two wall surfaces 175 constituting the lower portion 125 , thereby increasing the stability of the embedding container 100h when it is placed on the installation surface 180 .

[0137] Figure 8 In the embedding container 100g shown, the lower space of the lower portion 125 is located near the center of the boundary surface between the upper portion 120 and the lower portion 125. Therefore, the water absorbing member 190, which is a porous member, is located near the center of the boundary surface between the upper portion 120 and the lower portion 125. Figure 9 In the embedding container 100h shown, the lower space of the lower portion 125 is provided in a position close to the wall surface 175 of the embedding container 100h. Therefore, the water absorbing member 190 is provided in a position close to the wall surface 175 of the embedding container 100h. Figure 9 The step 173 of the embedding container 100 h shown is inclined toward the boundary between the upper portion 120 and the lower portion 125 .

[0138] Figure 9 The embedding container 100h shown has a slope formed by the step 173. Therefore, when the liquid 160 is introduced or discharged, even if the object 115 floats from the water absorbing member 190, the object 115 slides onto the water absorbing member 190 and can be collected on the water absorbing member 190.

[0139] Figure 10 Another example of the configuration of the embedding container 100 in this embodiment is shown. Figure 9 The wall surface 175 of the upper portion 120 of the embedding container 100h shown is flat. Figure 10 The embedding container 100i shown has a protrusion 176 on the wall surface 175 of the upper portion 120, and a water absorbing member 190 is provided between the boundary between the upper portion 120 and the lower portion 125 and the protrusion 176. It should be noted that a filter 118 may be provided in place of or in addition to the water absorbing member 190.

[0140] exist Figure 1 The embedding container 100 shown, Figure 9 In the embedding container 100h shown in FIG. 1 , the lower surface of the object 115 is placed on the filter 118, and Figure 10 In the embedding container 100i shown in FIG. 1 , the side surface of the object 115 is placed on the water absorbing member 190. Figure 10 The embedding container 100i shown can be manufactured with Figure 1 In the case of the embedding container 100 , etc., a cross-sectional specimen of the object 115 is cut at a different angle. Figure 10The embedding container 100i shown has a small lower space, so the consumption of the liquid 160 is small and economical.

[0141] Figure 11 Another example of the structure of the embedding container 100 in this embodiment is shown. Figure 1 In the embedding container 100 shown, the lower portion 125 is provided with a hole 130, and the introduction and discharge of the liquid 160 are both carried out through the hole 130. Figure 11 In the embedding container 100 j shown, in addition to the hole 130 , a passage 131 and a passage 132 communicating with the outside of the embedding container 100 are provided for the introduction and / or discharge of the liquid 160 .

[0142] Figure 11 The embedding container 100j shown can introduce liquid 160 through the passages 131 and 132 and discharge the liquid 160 through the hole 130. However, the present invention is not limited thereto, and any one of the hole 130, the passage 131, and the passage 132 may be used for introducing the liquid 160, and any one of them may be used for discharging the liquid 160.

[0143] Figure 11 In the embedding container 100 j shown, the liquid 160 is introduced from above the object 115 . Therefore, the object 115 is less likely to float when the liquid 160 is introduced, and the possibility of the object 115 being lost can be reduced. Figure 11 Since the embedding container 100j shown has a small lower space, when the embedding container 100j is cut from the bottom surface of the lower portion 125 toward the upper portion 120, the distance to the object 115 can be shortened, thereby reducing the number of cuts.

[0144] Figure 12 An example of the aggregation structure of the embedding container 100 in this embodiment is shown (perspective view). Figures 1 to 11 As shown, it can be used alone or as Figure 12 As shown, a gathering structure 101 is employed in which a plurality of embedding containers 100k are gathered together.

[0145] Figure 12 In the shown embedding container 100k, a plurality of upper spaces of the aggregation structure 101 are aggregated and arranged in a planar shape. Figure 12 The illustrated clustering structure 101 of the embedding containers 100k is a 6-hole clustering structure in which the embedding containers 100k are arranged in 2 rows and 3 columns, but is not limited thereto and may be a clustering structure in which 2 or 3 embedding containers 100k are arranged in a row, a 4-hole clustering structure in which they are arranged in 2 rows and 2 columns, a 12-hole clustering structure in which they are arranged in 3 rows and 4 columns, a 24-hole clustering structure in which they are arranged in 4 rows and 6 columns, or a 96-hole clustering structure in which they are arranged in 8 rows and 12 columns.

[0146] Figure 12 The illustrated clustering structure 101 of the embedding containers 100k can easily prepare specimens of the multiple objects 115 by clustering the multiple embedding containers 100k together to accommodate multiple objects 115 at once. The method for preparing specimens using the clustering structure 101 of the embedding containers 100k will be described later.

[0147] Figure 13 Another example of the aggregation structure of the embedding container 100 in this embodiment is shown (a diagram viewed from above). Figure 12 The illustrated aggregation structure 101 of the embedding containers 100k is an aggregation structure of cylindrical embedding containers 100k. Figure 13 The illustrated clustered structure 102 of the embedding containers 1001 is formed by clustering the hexagonal prism-shaped embedding containers 1001 in a closest-packed manner, with the upper spaces of the clustered containers arranged in a planar shape forming a honeycomb structure.

[0148] Figure 13 The clustering structure 102 of the embedding container 1001 shown has a honeycomb structure, which provides excellent strength and shock absorption. It can withstand the impact and vibration when the liquid 160 is introduced into and discharged from the embedding container 1001, the impact and vibration when the clustering structure 102 is cut, etc., and can prevent deformation of the embedding container. Figure 13 The aggregate structure 102 of the embedding container 1001 shown has a honeycomb structure, whereby the material of the embedding container 1001 can be saved, and the embedding container 1001 can be made lightweight while having sufficient strength.

[0149] Figure 13 The shown aggregation structure 102 of the embedding container 1001 has a hexagonal honeycomb structure. However, the embedding container 1001 may be not only a hexagonal prism, but may also be a densely packed aggregation structure of a triangular prism (or a triangular pyramid, a triangular pyramid, etc.) or a quadrangular prism (or a quadrangular pyramid, a quadrangular pyramid, etc.), or a densely packed aggregation structure in which a part of the hexagonal prism includes a pentagonal prism or other polygonal prism (or a polygonal pyramid, a polygonal pyramid, etc.) shape.

[0150] Figure 14 Another example of the structure of the embedding container 100 in this embodiment is shown (a view viewed from above). Figure 13 The embedding container 1001 shown is in the shape of a hexagonal prism. Figure 14 In the illustrated embedding container 100m, an outer peripheral portion 196 surrounding the embedding container 100m is in the shape of a hexagonal prism.

[0151] The embedding container 100m may further include an outer peripheral portion 196 surrounding the embedding container 100m. For example, the outer peripheral portion 196 may have a honeycomb structure surrounding the columnar embedding container 100m. The outer peripheral portion 196 may be formed of the same material as the embedding container 100m. The outer peripheral portion 196 may or may not be a polygonal prism (e.g., a polygonal pyramid, a polygonal pyramid, etc.). The embedding container 100m may also be secured to a known microtome via the outer peripheral portion 196.

[0152] The outer peripheral portion 196 may have a surface that is not perpendicular to the cutting direction of the object 115. For example, the non-perpendicular surface is an arc-shaped surface or a surface containing the vertices of a polygon. The outer peripheral portion 196 having a surface that is not perpendicular to the cutting direction of the object 115 can prevent deformation of the embedding container 100.

[0153] The outer peripheral portion 196 may include an intermediate portion 198 extending radially between the outer peripheral portion 196 and the embedding container 100m. The intermediate portion 198 is a region provided to mitigate pressure applied perpendicular to the cutting direction during cutting of the object 115. For example, the radial direction is the direction in which the embedding container 100m is cut. The intermediate portion 198 may be hollow or solid.

[0154] The distance from the end of the embedding container 100 to the end of the outer peripheral portion 196 can be defined as the short side v. For example, the intermediate portion 198 is a region where the short side v is between 0.2 mm and 10 mm. The intermediate portion 198 can be a region extending 3 mm or more along the direction in which the embedding container 100 is cut. Furthermore, the inclusion of the intermediate portion 198 in the outer peripheral portion 196 prevents deformation of the embedding container 100.

[0155] Figure 14 The embedding container 100m shown may not have a honeycomb structure. However, by providing the outer peripheral portion 196 to have a honeycomb structure, the embedding container 100m has excellent strength and shock absorption, and can prevent deformation of the embedding container.

[0156] Figure 15 An example of the configuration of an embedding device 800 in this embodiment is shown. The embedding device 800 embeds the object 115 and prepares a specimen. The embedding device 800 may include all or part of the embedding container 100, the moving unit 300, the biological manipulation unit 400, and the introduction unit 500.

[0157] The embedding container 100 provided in the embedding device 800 can be any of the embedding containers 100 of the previously described embodiments, depending on the purpose and application. The embedding device 800 may include only one embedding container 100, or may include a plurality of embedding containers 100 aggregated, as shown in the aggregate structure 101 or the aggregate structure 102.

[0158] The moving unit 300 moves the embedding container 100. The moving unit 300 can bring the embedding container 100 from outside the embedding apparatus 800 into the embedding apparatus 800, or can remove the embedding container 100 housed inside the embedding apparatus 800 and move it to an embedding dish located outside the embedding apparatus 800. The moving unit 300 is not limited to this, and can move the embedding container 100 from any location to any location depending on the intended purpose.

[0159] The moving unit 300 can receive input from the outside (such as an operator) and automatically move the embedding container 100 via an actuator. Alternatively, the embedding container 100 can be manually moved by an operator instead of the moving unit 300. In this case, the embedding device 800 does not need to include the moving unit 300.

[0160] The biological manipulation unit 400 manipulates the object 115. The biological manipulation unit 400 can accommodate the object 115 within the embedding container 100. For example, the biological manipulation unit 400 is a manipulator equipped with a nozzle for manipulating cells serving as the object 115. Operations of the biological manipulation unit 400 can be performed automatically based on input from an external source (e.g., an operator). The nozzle can have a flow path within it. The biological manipulation unit 400 can include a pump for supplying and / or drawing gas into the flow path of the nozzle, a support for moving the nozzle, and the like. Details of the manipulation of the object 115 by the nozzle will be described later.

[0161] The introduction unit 500 introduces the liquid 160 into the embedding container 100 and / or discharges the liquid 160 from the embedding container 100. For example, the introduction unit 500 introduces a treatment liquid for treating the object 115 into the embedding container 100. For example, the introduction unit 500 introduces an embedding agent for embedding the object 115 into the embedding container 100. Instead of using the introduction unit 500, an operator may manually introduce the liquid 160 into the embedding container 100 and / or discharge the liquid 160 from the embedding container 100. The introduction and / or discharge of the liquid 160 by the introduction unit 500 may be performed automatically in response to input from an external source (e.g., an operator).

[0162] The introduction section 500 may include a syringe for introducing a liquid into the embedding container 100. The introduction section 500 may include a manifold for accommodating the embedding container 100. In this case, the syringe and the manifold may have the function of introducing and / or discharging the treatment liquid and / or embedding agent into the embedding container 100, as will be described in detail later.

[0163] The processing liquid can be a liquid 160 that has the function of fixing, dehydrating and / or defatting the object 115. The organic solvent contained in the liquid 160 can be the processing liquid. The processing liquid can be one or more of formalin (formaldehyde, paraformaldehyde), glutaraldehyde, Bouin's preservative solution, Carnoy's solution, methanol, ethanol, xylene and xylene substitutes, but is not limited thereto. The embedding agent can be a substance for embedding the object 115. The organic solvent contained in the liquid 160 can be the embedding agent. For example, the embedding agent is paraffin or collodion. The processing liquid and / or the embedding agent can be used at any temperature depending on the purpose. As an example, formalin can be cooled to about 0°C for use, and paraffin can be heated to about 60°C for use.

[0164] In addition to the above, the embedding device 800 may also include a camera for capturing the process of preparing the specimen and a computer for analyzing the captured image files and video files, as will be described in detail later.

[0165] Figure 16 An example of the flow of the method for sampling the object 115 in this embodiment is shown. The object 115 in this embodiment can be sampled by Figure 16 It should be noted that, for ease of description, the processes of S100 to S900 are described in sequence, but at least a portion of these processes may be performed in parallel, or the steps may be performed alternately within the scope of the present invention.

[0166] First, in S100, the embedding device 800 places the object 115 in the embedding container 100. S100 can be referred to as a placement step. The step of placing the object 115 in the embedding container 100 in S100 is as follows: Figure 17 As shown, it may include steps S110 to S190.

[0167] Figure 17 S100 in the flow is shown. As a method for recovering the object 115, the method described in S110 to S150 is exemplified. However, as long as the object 115 can be recovered, the method is not limited to the method described in S110 to S150, and a known method may be used to recover the object 115.

[0168] First, in S110, the biological manipulation unit 400 forms an air bubble at the tip of the nozzle. The nozzle can manipulate an object 115 in the liquid 160 by passing fluid through the interior of the flow path. The flow path allows fluid to be drawn in (intake) or discharged (supply). The flow path is arranged to extend along the length of the nozzle. The nozzle can be a glass tube. For example, the glass tube can be formed using known glass forming techniques such as manual methods, the Danner method, and the bellows method.

[0169] The fluid can be a gas and / or a liquid. The gas can be air or contain water. The gas can be sterile. For example, by discharging gas from a nozzle into liquid 160 to form bubbles, the bubbles can be used to manipulate the object 115 in liquid 160. The flow path can pass through the object 115 along with the fluid.

[0170] Figure 18 and Figure 19 1 is a diagram for explaining each step from S110 to S190 of arranging the object 115 inside the embedding container 100 as the arranging step of S100. Figure 18 In S110, the length of the nozzle 49 in the longitudinal direction may be greater than 1 mm and less than 50 mm. The length of the nozzle 49 in the longitudinal direction may be greater than 5 mm and less than 30 mm. The length of the nozzle 49 in the longitudinal direction may be greater than 10 mm and less than 20 mm.

[0171] The inner diameter of flow path 51 can be between 1 μm and 10,000 μm. The inner diameter of flow path 51 can be between 10 μm and 1,000 μm. The inner diameter of flow path 51 can be between 20 μm and 500 μm. When the inner diameter of flow path 51 and / or the longitudinal length of nozzle 49 are within the above ranges, nozzle 49 can retain a certain amount of fluid, thereby facilitating manipulation of object 115. Furthermore, excessively long nozzle 49 does not make it difficult to guide tip 254 of nozzle 49 to a desired position.

[0172] exist Figure 18 and Figure 19 In the example of the object 115, adherent cells are shown, but the object 115 may also be suspended cells. In addition, the object may also be a single cell or a cell aggregate such as a sphere. Figure 18 In S110, adherent cells are cultured in a solid phase located on the inner bottom surface of a cell culture container 250. The adherent cells can be immersed in a liquid 160 in the cell culture container 250. The liquid 160 can be a complete culture medium, a basal culture medium, or a buffer solution. It should be noted that when the object 115 is a non-living organism or a dead cell, the liquid 160 is not limited thereto and can also be water, an organic solvent, or the like.

[0173] exist Figure 18 In S110, a pump (not shown) connected to the end 253 of the nozzle 49 opposite to the tip 254 supplies gas to the flow path 51 of the nozzle 49. This forms bubbles 256 at the tip 254. The bubbles 256 formed and maintained at the tip 254 enable manipulation of adherent cells.

[0174] Next, in S130, the biological manipulation unit 400 brings the bubble 256 formed at the front end 254 of the nozzle 49 into contact with the adherent cells, causing the adherent cells to adhere to the gas-liquid interface 255 of the bubble 256. By controlling the pump, the pressure and / or volume of the gas supplied or inlet can be controlled. Thus, since the size of the bubble 256 can be varied, adherent cells within a desired range can be detached. It should be noted that, instead of moving the nozzle 49 along the surface of the solid phase, the cell culture container 250 can be moved to detach the adherent cells from the solid phase.

[0175] Next, in S150, the pump draws gas from the flow channel 51 of the nozzle 49, thereby recovering adherent cells attached to the bubbles 256 within the flow channel 51. In this manner, the bubbles 256 formed at the tip 254 can be used to selectively detach adherent cells from the solid phase and recover them within the flow channel 51. By supplying and aspirating gas into the flow channel 51 of the nozzle 49, the nozzle 49 facilitates manipulation of adherent cells within the liquid 160. This method allows for the recovery of the object 115 without excessively aspirating liquid 160 other than the object 115. Therefore, the amount of liquid 160 contained in the embedding container 100 along with the object 115 can be reduced compared to conventional pipetting. Cell manipulation can include recovering aggregates of cells from single cells by changing the nozzle diameter, transferring cells while they are attached to the air-liquid interface 255 of the bubbles 256, and at least some of the following: cell attachment, detachment, and squeezing using bubbles.

[0176] In the above description, the case where the object 115 is an adherent cell is described. However, when the object 115 is a suspended cell, the suspended cell can also be brought into contact with the bubble 256 formed at the front end 254 of the nozzle 49, so that it adheres to the gas-liquid interface 255 and is recovered, or the suspended cell can be recovered by other well-known methods.

[0177] Next, in Figure 19 In S170, the support unit moves the nozzle 49 onto the embedding container 100, and positions the object 115 within the embedding container 100 (e.g., onto the filter 118 or onto the water-retaining and absorbing member 190). When the object 115 is positioned on the filter 118 or the water-retaining and absorbing member 190, the filter 118 or the water-retaining and absorbing member 190 is embedded while in contact with the object 115. A pump supplies gas to the flow path 51 of the nozzle 49, thereby forming bubbles 256 at the tip 254 of the nozzle 49 to allow adherent cells to attach. The support unit can bring the bubbles 256 of the nozzle 49 into contact with the liquid 160 within the embedding container 100 and / or insert the bubbles 256 into the liquid 160, thereby positioning the object 115 within the embedding container 100.

[0178] exist Figure 19 , a configuration is shown in which no lid is provided in the upper space of the embedding container 100. However, a configuration may also be employed in which a hole of a size that allows a nozzle 49 or a dispenser (e.g., a pipette) for placing the object 115 into the interior of the embedding container 100 to pass is provided in the lid of the embedding container 100, and the nozzle 49 or the like can be inserted into the upper space of the embedding container 100 through the hole.

[0179] Next, in S190, the moving unit 300 moves the embedding container 100 containing adherent cells and stores it in the manifold 510 of the introduction unit 500. Instead of the moving unit 300 moving the embedding container 100, the operator may manually store the embedding container 100 in the manifold 510. After the embedding container 100 is stored in the manifold 510, the embedding device 800 proceeds to S200. Figure 19 While the figure shows an embodiment in which there is no step 173 at the boundary between the upper portion 120 and the lower portion 125 of the embedding container 100, a step 173 may be provided at the boundary between the upper portion 120 and the lower portion 125 of the embedding container 100. The step 173 of the embedding container 100 may correspond to the step arrangement of the manifold described later, thereby making it easier to determine the height during storage.

[0180] Next, in S200, the embedding device 800 pre-treats the object 115 placed in the embedding container 100 using a treatment liquid. The embedding device 800 can pre-treat the object 115 while the object 115 is placed in the embedding container 100. S200 can be a pre-treating step. The pre-treating step can be fixing, dehydrating, and / or defatting the object 115. The treatment liquid can be a fixing liquid, a dehydrating liquid, and / or a defatting liquid. The steps for pre-treating the object 115 in S200 are as follows: Figure 20 As shown, steps S210 to S250 may be included.

[0181] Figure 20 S200 in the flow chart is shown. First, in S210, the introduction unit 500 introduces a fixative into the embedding container 100 to fix the object 115. The fixative can be formalin, glutaraldehyde, Bouin's solution, Carnoy's solution, or methanol. A cooled substance (e.g., formalin cooled to approximately 0°C) can be used as the fixative.

[0182] Figure 21 1 is a diagram illustrating a state in which the introduction unit 500 introduces a liquid 160 such as a fixing solution into the embedding container 100 as a pre-processing step. Figure 21In the process, the embedding container 100 is housed in the manifold 510. The hole 130 in the lower portion 125 of the embedding container 100 is connected to and communicates with the passage 231 of the manifold 510. The introduction unit 500 opens the valve 261 and closes the valve 262. Next, the introduction unit 500 places the syringe 530 in the upper space of the upper portion 120 of the embedding container 100 and pulls the syringe 530 upward (away from the embedding container 100), thereby applying negative pressure to the upper and lower spaces of the embedding container 100. As a result, the fixative is introduced from the fixative container, which serves as the processing liquid supply container 240, through the passage 232 and the passage 231 into the embedding container 100. After an amount of fixative sufficient to fix the object 115 has been introduced into the embedding container 100, the introduction unit 500 stops pulling the syringe 530 upward.

[0183] After a sufficient time has passed to fix the object 115, the introduction unit closes valve 261 and opens valve 262. Next, the introduction unit 500 presses the syringe 530 downward (toward the embedding container 100), thereby applying positive pressure to the upper and lower spaces of the embedding container 100. As a result, the fixative solution that fills the embedding container 100 is discharged into the waste liquid container 246 through the passage 231 and the passage 233. After all or a portion of the fixative solution (e.g., half, three-quarters, or the entire amount after leaving enough to immerse the object 115) has been discharged from the embedding container 100, the introduction unit 500 stops pressing the syringe 530 downward.

[0184] If the fixation of the object 115 is insufficient, the operation of introducing the fixative into and draining the embedding container 100 may be repeated two or more times. Figure 21 In the figure, a method using a syringe 530 is shown, but a method of applying negative pressure and / or positive pressure to the upper and lower spaces of the embedding container 100 by using a pump for sucking and supplying gas to the upper space of the upper portion 120 of the embedding container 100 may be used instead of the syringe 530. Figure 21 , a method is shown in which the hole 130 of the lower portion 125 of the embedding container 100 is connected and communicated with the passage 231 of the manifold 510. However, a method may also be used in which the liquid 160 is introduced into the upper space and the lower space of the embedding container 100 from an opening of the upper portion 120 of the embedding container 100, where the hole 130 is not provided, using a nozzle 49 for sucking and supplying the liquid 160.

[0185] Next, in S230, the introduction unit 500 introduces a dehydrating liquid into the embedding container 100 to dehydrate the object 115. The dehydrating liquid can be an alcohol such as ethanol, and any concentration of ethanol, such as absolute ethanol, 90% ethanol, 80% ethanol, or 70% ethanol, can be used. The introduction and removal of the dehydrating liquid into and from the embedding container 100 can be performed using the same steps as S210. In this case, the processing liquid supply container 240 can use a container for the dehydrating liquid, and multiple containers can be provided to accommodate the different ethanol concentrations and applications. Step S230 can also be performed multiple times, as needed, to introduce and remove the dehydrating liquid.

[0186] Next, in S250, the introduction unit 500 introduces a degreasing liquid into the embedding container 100 to degrease the object 115. The degreasing liquid can be xylene. Since ethanol, the dehydrating liquid, is incompatible with paraffin, the embedding medium, xylene can be used for treatment between dehydration and embedding. The introduction and removal of the degreasing liquid into and from the embedding container 100 can be performed using the same steps as S210. In this case, a container for the degreasing liquid can be used as the processing liquid supply container 240. Step S250 can also be performed multiple times, as needed, to introduce and remove the degreasing liquid.

[0187] In the case of the object 115 rich in fat, a defatting treatment may be performed before dehydration. In the case of the object 115 being hard tissue, a decalcification treatment using an acid, a chelating agent, etc. may be performed before dehydration. After the pretreatment of the object 115, the embedding device 800 proceeds to step S300.

[0188] Next, in S300, the embedding device 800 embeds the object 115. The embedding device 800 can embed the object 115 together with the embedding container 100, that is, together with the holding member. This operation can be referred to as an embedding step. As an example, the embedding device 800, with the object 115 already placed in the embedding container 100, introduces an embedding agent into the embedding container 100, and embeds the object 115 within the embedding container 100. This operation can also be referred to as an in-container embedding step.

[0189] Figure 22FIG2 is a diagram of an embedding container 100 filled with an embedding agent. Specifically, in the embedding step, the embedding apparatus 800 uses a container containing an embedding agent (e.g., paraffin wax 162 or collodion) as a processing liquid container and fills the embedding container 100 with the embedding agent through the same steps as S210. A heated embedding agent (e.g., paraffin wax at approximately 60°C) can be used. The upper and lower spaces of the embedding container 100 can be completely filled with the embedding agent, and at least a portion of the upper space can be filled with the embedding agent until the object 115 is fully immersed. The embedding container 100 can be left to stand until the embedding agent has sufficiently cooled and solidified.

[0190] Figure 23 The figure shows an embedding dish 280 containing an embedding container 100 filled with paraffin wax 162 as an embedding agent. In this case, the embedding agent not only fills the interior of the embedding container 100 but also surrounds its periphery. Specifically, as an embedding step, the embedding apparatus 800 can place the embedding container 100 in the embedding dish 280 and attach the embedding cassette 290 to the top of the embedding dish 280. The embedding container 100 can be placed in the embedding dish 280 using the moving unit 300 or manually by an operator. The embedding container 100 placed in the embedding dish 280 can be the embedding container 100 before being filled with the embedding agent, or it can be the embedding container 100 after being filled with the embedding agent through the in-container embedding step. The embedding agent can be filled into the embedding dish 280 manually by an operator. The embedding dish 280 may be filled with the embedding agent by the embedding apparatus 800 receiving input from the outside (an operator, etc.) and automatically filling the embedding dish 280 in which the embedding container 100 is placed with the embedding agent.

[0191] Next, in S400 , the object 115 is sliced ​​by slicing the embedding container 100 that has undergone the intra-container embedding step and / or the embedding step.

[0192] Figure 24 This figure explains the process of slicing the embedding container 100. The block of embedding medium embedded in the embedding container 100 can be taken out from the embedding dish 280. The taken-out block of embedding medium can be placed with the embedding cassette 290 facing downward.

[0193] Next, the embedding block can be sliced ​​using a blade 295. At this point, Figure 24As shown, since object 115 is located directly below filter 118, until filter 118 is visually observed on the sliced ​​cross section, it indicates that object 115 is absent, allowing for rough sectioning. Once filter 118 appears on the cut surface, it indicates the presence of object 115 nearby, allowing for careful sectioning. Repeated sectioning yields a thin slice containing object 115 embedded in the embedding agent (referred to as the first small slice) and a thin slice containing filter 118 embedded in the embedding agent (referred to as the second small slice). The first and second small slices are close (adjacent) within the embedding agent block. Therefore, after obtaining the second small slice during sectioning, careful sectioning can be performed to obtain the first small slice. In this manner, by adjusting the sectioning of the embedding agent block containing embedding container 100, using the presence of filter 118 (i.e., the second small slice) as a criterion, a specimen of object 115 (i.e., the first small slice) can be easily prepared without losing object 115.

[0194] As the cutting target, as described above, instead of the presence of the filter 118, / on this basis, one or more of the value of the cross-sectional area of ​​a predetermined cross section, the presence of the step 173, the presence of the water-absorbing component 190, and the color switching of the water-absorbing component 190 can be used as the target.

[0195] The embedding container 100 can be sectioned using a microtome or other device, or it can be manually performed by an operator. Sectioning of the embedding container 100 can also be performed automatically. For example, the embedding apparatus 800 automatically captures the sectioning process of the embedding container 100 in a time series using a camera included in the embedding apparatus 800. The embedding apparatus 800 can perform image analysis on the cross-sectional images of the embedding container 100 captured in a time series using a computer included in the embedding apparatus 800.

[0196] The computer included in the embedding apparatus 800 can analyze a plurality of images captured in a time series and control the blade 295 to roughly cut the embedding container 100 before a cross-sectional image satisfies predetermined conditions (for example, the filter 118 appears on the cross section or the cross-sectional area of ​​the container is greater than / less than a threshold value, etc.), and to carefully cut the embedding container 100 after the filter 118 appears on the cross section.

[0197] "Rough cutting" may be performed with a relatively thick thickness (e.g., more than 10 μm, more than 15 μm, more than 20 μm, more than 50 μm, or more than 100 μm). "Careful cutting" may be performed with a relatively thin thickness compared to "rough cutting," for example, less than 1 μm, less than 2 μm, less than 3 μm, less than 4 μm, less than 5 μm, less than 6 μm, less than 7 μm, less than 8 μm, less than 9 μm, or less than 10 μm.

[0198] "Rough cutting" means that the cutting speed is relatively fast (for example, the number of specimens cut in 1 minute is more than 5, 6, 7, 8, 9, or 10). "Careful cutting" means that the cutting speed is relatively slow compared to "rough cutting" (for example, the number of specimens cut in 1 minute is less than 4, 3, 2, or 1).

[0199] For example, the computer included in the embedding apparatus 800 calculates the cross-sectional area values ​​of the cross sections appearing in the plurality of images captured in time series and controls the blade 295 to roughly cut the embedding container 100 before the cross-sectional area value of the cross section is reached, and then carefully cut the embedding container 100 when the cross-sectional area value of the cross section is reached. For example, the computer included in the embedding apparatus 800 analyzes the plurality of images captured in time series and controls the blade 295 to roughly cut the embedding container 100 before the water absorbing member 190 appears in the cross section, and then carefully cut the embedding container 100 after the water absorbing member 190 appears in the cross section. For example, the computer included in the embedding apparatus 800 analyzes the plurality of images captured in time series and controls the blade 295 to roughly cut the embedding container 100 before the color of the water absorbing member 190 appears in the cross section changes, and then carefully cut the embedding container 100 after the color of the water absorbing member 190 appears in the cross section changes.

[0200] When the operator manually sections the embedding container 100, the computer included in the embedding apparatus 800 can notify the operator, via the display included in the embedding apparatus 800, that the filter 118 has appeared on the cross section. When the operator manually sections the embedding container 100, the computer included in the embedding apparatus 800 can also notify the operator, via the display included in the embedding apparatus 800, that the cross-sectional area value has reached a predetermined value, that a step difference 173 has appeared, that the water absorbing member 190 has appeared, and / or that the color of the water absorbing member 190 has changed, but the present invention is not limited thereto. In this way, the embedding apparatus 800 can provide the operator with a standard for cutting the embedding container 100.

[0201] The computer analysis may be performed by a program having internal instructions. The instructions may include instructions that, when executed by a processor or a programmable circuit, cause the processor or the programmable circuit to perform the analysis.

[0202] Next, in S500, the small piece (specimen) containing the object 115 obtained by sectioning is stretched. The specimen can be stretched by transferring it to a water bath and floating it on the water. If the stretching is insufficient, the specimen will wrinkle, which is not desirable. After the stretching is complete, the specimen can be placed on a glass slide and dried.

[0203] Next, in S600, the specimen is deparaffinized. If deparaffinization is insufficient, the specimen will contain paraffin and will not be adequately stained. Deparaffinization can be performed using known methods. For example, deparaffinization can be performed by sequentially immersing a slide containing the specimen in glass bottles filled with xylene, 100% ethanol, and 70% ethanol, respectively.

[0204] Next, in S700, the specimen is stained. Specimens can be stained using known staining methods such as hematoxylin and eosin staining (HE staining), Cuban-Barrel staining (KB staining), melanin staining, Giemsa staining, Gram staining, or toluidine blue staining. For example, HE staining is performed by sequentially immersing the deparaffinized specimen in glass bottles filled with hematoxylin and eosin solutions, respectively.

[0205] Next, in S800, the specimen is dehydrated and cleared. The specimen can be dehydrated and cleared by known methods. For example, the specimen can be dehydrated and cleared by sequentially immersing the stained specimen in glass bottles filled with 70% ethanol, 100% ethanol, and xylene, respectively.

[0206] Next, in S900, the specimen is enclosed on a glass slide. The specimen can be enclosed using known methods. For example, the specimen can be enclosed by placing an encapsulant on a glass slide carrying a dehydrated and transparent specimen, and then placing a cover glass on top. The encapsulant can contain glycerin, or a commercially available product can be used. The periphery of the cover glass can be sealed with commercially available nail polish, etc. The glass slide produced in this manner through steps S100 to S900 can be used for microscopic observation, etc.

[0207] Next, a modified example of the method of sampling the object 115 is shown. Figure 12 and Figure 13 The illustrated sample shows the aggregate structure of the embedding container 100 in which the object 115 is embedded.

[0208] (Variation 1)

[0209] Figure 25 The modified example 1 shown is a method in which an object placed on a cell culture plate 103 is stored as a specimen on a single slide glass 104 . Figure 25 Although the cell culture plate 103 is shown as having 96 wells, it is not limited thereto and may be a cell culture plate having 6 wells, 12 wells, or 24 wells.

[0210] Figure 26This figure illustrates steps S170 and S190 in Modification 1. In Modification 1, steps S110 to S150 can be performed using the method already described. In S170, the support unit moves the nozzle 49 onto the embedding container cassette 108 and places the object 115 inside the embedding container cassette 108 (e.g., on the filter 118). The embedding container cassette 108 can be a 96-well type embedding container assembly. The size of the embedding container cassette 108 can be such that it fits within the glass slide 104. For example, if the glass slide 104 is approximately 26 mm long and 76 mm wide, the embedding container cassette 108 can be approximately 24 mm long and 37 mm wide.

[0211] In S170, gas is supplied to the flow path 51 of the nozzle 49 by a pump, thereby forming bubbles 256 at the tip 254 of the nozzle 49 to which the objects 115 adhere. The support unit allows the bubbles 256 of the nozzle 49 to contact and / or penetrate the liquid 160 within the embedding cartridge 108, thereby placing the objects 115 within any well of the embedding cartridge 108. The objects 115 cultured (or immersed) within the cell culture plate 103 can be placed in corresponding wells of the embedding cartridge 108.

[0212] Next, in S190, the moving unit 300 moves the embedding container cassette 108 containing the object 115 and stores it in the manifold 510a of the introduction unit 500a. Instead of using the moving unit 300 to move the embedding container cassette 108, an operator may manually store the embedding container cassette 108 in the manifold 510a. Here, the manifold 510a and syringe 530a may be of a collective structure.

[0213] For example, the manifold 510a and the syringe 530a are arranged with 96 (8 rows and 12 columns) Figure 21 The structure of the manifold 510 and syringe 530 is shown. However, the manifold 510a and syringe 530a may be configured to correspond to the cell culture plate 103. After the embedding container 108 is placed in the manifold 510a, the embedding apparatus 800 proceeds to step S200. Steps after S200 can be performed using the previously described method.

[0214] exist Figure 25 and Figure 26 In the first modification shown, all objects 115 are collected on one well plate and arranged in an array pattern on the slide 104 . Thus, specimens of the objects 115 are collected on one slide 104 , enabling observation of the specimens at once, which is very convenient.

[0215] (Variation 2)

[0216] Figure 27 The second modification shown is a mode in which the objects arranged in the embedding containers 106 included in the aggregation structure 105 are individually housed on a slide glass 107 . Figure 27 Although the case where the clustering structure 105 has 96 wells (96 embedding containers 106 ) is shown, the structure is not limited thereto and may have 6 wells, 12 wells, 24 wells, or the like.

[0217] Figure 28 An example of a cross-sectional view of each embedding container 106 included in the aggregation structure 105, taken along a cross section perpendicular to the installation surface 180, is shown. Each embedding container 106 has a hole 130 in the lower portion 125, a filter 118 is provided at the boundary between the upper portion 120 and the lower portion 125, and an object 115 is placed on the filter 118. The object 115 can be cultured within the embedding container 106, which is filled with a culture medium as a liquid 160.

[0218] Figure 29 FIG2 is a diagram illustrating steps S100 to S270 in Modification 2. In Modification 2, since the aggregation structure 105 includes each embedding container 106, when the object 115 is cultured (or immersed) in each embedding container 106, it is not necessary to perform Figure 17 The operations from S110 to S170 are shown.

[0219] First, in S190 , the moving unit 300 moves the collecting structure 105 in which the objects 115 are arranged, and accommodates the collecting structure 105 in the manifold 510 b of the introduction unit 500 b .

[0220] Figure 30 1 is a diagram illustrating step S190 in modification 2. In step S190, the moving unit 300 moves the collecting structure 105 on which the object 115 is arranged and accommodates it in the manifold 510b of the introduction unit 500b. For example, step S190 is similar to Figure 26 Step S190 of the first modification shown is performed similarly. Instead of moving the collecting structure 105 using the moving unit 300, the operator can manually place the collecting structure 105 in the manifold 510b. Here, the manifold 510b and the syringe 530b can be the manifold and syringe of the collecting structure, respectively. After the collecting structure 105 is placed in the manifold 510b, the embedding device 800 proceeds to step S200.

[0221] Step S200 can be performed according to the method already described. Next, in S270, the embedding device 800 disassembles the aggregate structure 105 into the individual embedding containers 106. For example, the aggregate structure 105 may have perforations formed between the embedding containers 106, and the perforations may be used to disassemble the aggregate structure 105 into the individual embedding containers 106. If perforations are not formed in the aggregate structure 105, the aggregate structure 105 may be disassembled into the individual embedding containers 106 using scissors or the like. After S270, the steps from S300 onwards can be performed for each embedding container 106 according to the method already described.

[0222] exist Figures 27 to 30 In the second modification shown, the embedding containers 106 form a collection structure 105, allowing for sequential culture and pretreatment with the collection structure 105, thus simplifying operation. In the second modification, a single slide is used to prepare a specimen for each object 115 placed in each embedding container 106. However, a plurality of objects 115 may be collected on a single slide to prepare a specimen.

[0223] As another embodiment of the present invention, a cutting method is disclosed, wherein, in step S400 of slicing the object 115 by cutting the embedding container 106, rough cutting is performed before the target feature appears on the cut surface, and careful cutting is performed after the target feature appears on the cut surface. Here, rough cutting may refer to a relatively thick cut thickness (e.g., exceeding 10 μm, exceeding 15 μm, exceeding 20 μm, exceeding 50 μm, or exceeding 100 μm), and careful cutting may refer to a relatively thin cut thickness (e.g., 1 μm or less, 2 μm or less, 3 μm or less, 4 μm or less, 5 μm or less, 6 μm or less, 7 μm or less, 8 μm or less, 9 μm or less, or 10 μm or less) compared to rough cutting.

[0224] Rough cutting refers to a relatively fast cutting speed (for example, cutting 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, or 10 or more specimens in 1 minute), while careful cutting may refer to a relatively slow cutting speed compared to rough cutting (for example, cutting 4 or less, 3 or less, 2 or less, or 1 or less specimens in 1 minute). The standard feature may be one or more of the following: the presence of filter 118, the value of the cross-sectional area of ​​a predetermined cross section, the presence of step 173, the presence of water absorbing member 190, or the color change of water absorbing member 190.

[0225] As another embodiment of the present invention, a cutting method is disclosed in which, in step S400 of cutting the embedding container 106 to slice the object 115, a cross section of the embedding container 100 appears on the cut surface in addition to the object 115. In the present invention, since the object 115 is cut together with the embedding container 100, a cross section of the embedding container 100 appears on the cut surface. The cross section of the embedding container 100 can be a closed shape (e.g., a circle without a cut) or an open shape (e.g., a circle with a cut in part).

[0226] Figure 31 An example of the hardware configuration of the computer 1900 of the embedding apparatus 800 performing image analysis in step S400 is shown. The computer 1900 of this embodiment includes a CPU 2000, a CPU peripheral unit including a RAM 2020, a graphics controller 2075, and a display device 2080, all interconnected via a main controller 2082, a communication interface 2030 connected to the main controller 2082 via an input / output controller 2084, an input / output unit including a hard disk drive 2040 and a CD-ROM drive 2060, and a conventional input / output unit including a ROM 2010, a floppy disk drive 2050, and an input / output chip 2070 connected to the input / output controller 2084.

[0227] The main controller 2082 connects the RAM 2020 to the CPU 2000 and the graphics controller 2075, which access the RAM 2020 at a high transfer speed. The CPU 2000 operates based on programs stored in the ROM 2010 and RAM 2020, controlling various components. The graphics controller 2075 acquires image data generated by the CPU 2000 and other devices in a frame buffer located within the RAM 2020 and displays it on the display device 2080. Alternatively, the graphics controller 2075 may internally include a frame buffer for storing image data generated by the CPU 2000 and other devices. The generated various information (e.g., images of cut surfaces, etc.) can be displayed on the display device 2080.

[0228] The input / output controller 2084 connects the main controller 2082 to the communication interface 2030, which serves as a relatively high-speed input / output device, the hard disk drive 2040, and the CD-ROM drive 2060. The communication interface 2030 communicates with other devices via a network, either wired or wirelessly. Furthermore, the communication interface functions as hardware for communication. The hard disk drive 2040 stores programs and data used by the CPU 2000 within the computer 1900. The CD-ROM drive 2060 reads programs and data from the CD-ROM 2095 and provides them to the hard disk drive 2040 via the RAM 2020.

[0229] The input / output controller 2084 is also connected to the ROM 2010, the floppy disk drive 2050, and the relatively low-speed input / output devices of the input / output chip 2070. The ROM 2010 stores a startup program executed when the computer 1900 starts up and / or programs that depend on the hardware of the computer 1900. The floppy disk drive 2050 reads programs and data from the floppy disk 2090 and provides them to the hard disk drive 2040 via the RAM 2020. The input / output chip 2070 connects the floppy disk drive 2050 to the input / output controller 2084 and connects various input / output devices to the input / output controller 2084 via, for example, a parallel port, a serial port, a keyboard port, and a mouse port.

[0230] The program provided to the hard disk drive 2040 via the RAM 2020 is stored in a recording medium such as the floppy disk 2090, CD-ROM 2095, or IC card and provided by the user. The program is read from the recording medium, installed in the hard disk drive 2040 in the computer 1900 via the RAM 2020, and executed by the CPU 2000.

[0231] The program installed in the computer 1900 and causing the computer 1900 to function as an image analysis computer in the embedding apparatus 800 can be executed by the CPU 2000 or the like to function as the computer of the embedding apparatus 800 .

[0232] The information processing described in these programs is loaded into the computer 1900, whereupon the software functions as a specific mechanism, namely, an image analysis unit, operating in conjunction with the various hardware resources described above. Furthermore, these specific mechanisms implement calculations or processing of information corresponding to the intended use of the computer 1900 in this embodiment, thereby constructing a computer system unique to the embedding device 800 corresponding to the intended use.

[0233] For example, when computer 1900 communicates with an external device, CPU 2000 executes a communication program loaded into RAM 2020 and instructs communication interface 2030 to perform communication processing based on the processing described in the communication program. Under the control of CPU 2000, communication interface 2030 reads transmission data stored in a transmission buffer area on a storage device such as RAM 2020, hard disk drive 2040, floppy disk 2090, or CD-ROM 2095 and transmits the data to the network, or writes received data from the network to a receive buffer area on the storage device. In this manner, communication interface 2030 can also transmit and receive data to and from the storage device using DMA (Direct Memory Access). Alternatively, CPU 2000 can transmit and receive data by reading data from a storage device or communication interface 2030 at the source and writing the data to communication interface 2030 or a storage device at the destination.

[0234] Furthermore, the CPU 2000 reads all or a required portion of files or databases stored in external storage devices such as the hard disk drive 2040, CD-ROM drive 2060 (CD-ROM 2095), and floppy disk drive 2050 (floppy disk 2090) into the RAM 2020 through DMA transfer or the like, and performs various processes on the data in the RAM 2020. The CPU 2000 then writes the processed data back to the external storage device through DMA transfer or the like. During such processing, the RAM 2020 is considered to temporarily store the contents of the external storage device. In this embodiment, the RAM 2020 and the external storage device are collectively referred to as memory, recording unit, or storage device.

[0235] Here, a storage device or the like stores information required for information processing of image analysis, such as moving image data, as needed, and supplies it to each component of the computer of the embedding device 800 as needed.

[0236] In this embodiment, various programs, data, tables, databases, and other information are stored in such a storage device and become the subject of information processing. It should be noted that the CPU 2000 can also store a portion of the RAM 2020 in a cache memory and read and write to the cache memory. In this manner, the cache memory also assumes a portion of the functions of the RAM 2020. Therefore, in this embodiment, except for the case of distinguishing marks, the cache memory is also included in the RAM 2020, memory, and / or storage device.

[0237] Furthermore, the CPU 2000 performs various processing specified by the program's instruction sequence, including various calculations described in this embodiment, information processing, conditional judgments, and information retrieval / replacement, on the data read from the RAM 2020, and writes the data back to the RAM 2020. For example, when performing conditional judgments, the CPU 2000 compares the various variables described in this embodiment with other variables or quantities to determine whether conditions such as greater than, less than, greater than, less than, or equal to are met. If the conditions are met (or not met), the CPU 2000 branches to a different instruction sequence or calls a subroutine.

[0238] Furthermore, the CPU 2000 can search for information stored in a file or database within a storage device. For example, if the storage device stores multiple entries in which attribute values ​​of a second attribute are associated with attribute values ​​of a first attribute, the CPU 2000 can search the multiple entries stored in the storage device for an entry that meets the conditions specifying the attribute value of the first attribute and read the attribute value of the second attribute stored in the entry, thereby obtaining the attribute value of the second attribute associated with the first attribute that meets the specified conditions.

[0239] The program shown above can also be stored in an external recording medium. As a recording medium, in addition to the floppy disk 2090 and CD-ROM 2095, optical recording media such as DVDs or CDs, optical magnetic recording media such as MOs, magnetic tape media, semiconductor memories such as IC cards, etc. can be used. In addition, a storage device such as a hard disk or RAM provided in a server system connected to a dedicated communication network or the Internet can also be used as a recording medium, and the program can be provided to the computer 1900 via the network.

[0240] In this disclosure, the computer in embedding device 800 is shown as having a CPU 2000 as a processor, but the type of processor is not particularly limited. For example, a GPU, ASIA, FPGA, etc. can be used as the processor as appropriate. Furthermore, in this disclosure, the computer for image analysis in embedding device 800 is shown as having a hard disk drive 2040 as an auxiliary storage device, but the type of auxiliary storage device is not particularly limited. For example, other storage devices such as solid-state hard drives can be used in place of or in addition to hard disk drive 2040.

[0241] While the present invention has been described above using embodiments, the technical scope of the present invention is not limited to the scope described in the above embodiments. It is known to those skilled in the art that various modifications or improvements can be made to the above embodiments. As can be seen from the claims, embodiments incorporating such modifications or improvements are also encompassed within the technical scope of the present invention.

[0242] It should be noted that the order of execution of each process, such as actions, sequences, steps, and stages, in the apparatus and method described in the claims, specifications, and drawings, can be implemented in any order, unless otherwise specified, such as "before," "before," and the output of the pre-processing is used in the post-processing. Even if the action flow in the claims, specifications, and drawings is described using the phrases "first," "next," etc. for convenience, it does not necessarily mean that it must be implemented in that order.

[0243] Description of Reference Numerals

[0244] 49 nozzles

[0245] 51 flow path

[0246] 100 embedding containers

[0247] 100a embedding container

[0248] 100b embedding container

[0249] 100c embedding container

[0250] 100d embedding container

[0251] 100e embedding container

[0252] 100f embedding container

[0253] 100g embedding container

[0254] 100h embedding container

[0255] 100i embedding container

[0256] 100j embedding container

[0257] 100k embedding container

[0258] 100L embedding container

[0259] 100m embedding container

[0260] 101 Aggregate Structure

[0261] 102 Aggregate Structure

[0262] 103 cell culture plate

[0263] 104 slides

[0264] 105 Aggregate Structure

[0265] 106 embedding container

[0266] 107 slides

[0267] 108 embedding container box

[0268] 110 container body

[0269] 115 objects

[0270] 118 filters

[0271] 118a filter

[0272] 118b filter

[0273] 120 upper part

[0274] 125 lower part

[0275] 130 holes

[0276] 131 access

[0277] 132 channels

[0278] 160 liquid

[0279] 162 paraffin

[0280] Around 170

[0281] 172 protrusions

[0282] 173 levels

[0283] 175 wall

[0284] 176 salient

[0285] 178 cover

[0286] 180 setting surface

[0287] 190 water-absorbing parts

[0288] 196 Peripheral Department

[0289] 198 Middle Section

[0290] 231 access

[0291] 232 access

[0292] 233 access

[0293] 240 treatment liquid supply container

[0294] 246 waste liquid container

[0295] 250 cell culture vessels

[0296] 253 end

[0297] 254 front end

[0298] 255 gas-liquid interface

[0299] 256 bubbles

[0300] 261 valve

[0301] 262 valve

[0302] 280 embedding dish

[0303] 290 embedding cassette

[0304] 295 blade

[0305] 300 Mobile Department

[0306] 400 Biological Operation Department

[0307] 500 Import Department

[0308] 500a introduction section

[0309] 500b import section

[0310] 510 manifold

[0311] 510a manifold

[0312] 510b manifold

[0313] 530 syringe

[0314] 530a syringe

[0315] 530b syringe

[0316] 800 embedding device

[0317] 1900 Computer

[0318] 2000CPU

[0319] 2010ROM

[0320] 2020RAM

[0321] 2030 Communication Interface

[0322] 2040 hard drive

[0323] 2050 floppy disk drive

[0324] 2060 CD-ROM drive

[0325] 2070 input and output chip

[0326] 2075 graphics controller

[0327] 2080 display device

[0328] 2082 main controller

[0329] 2084 Input and Output Controller

[0330] 2090 floppy disk

[0331] 2095CD-ROM.

Claims

1. An embedding container for accommodating an object to be embedded, wherein the embedding container is characterized in that: A holding member is provided for holding the object and allowing an embedding medium for embedding the object to pass through. The holding member is embedded in the embedding agent together with the object.

2. The embedding container according to claim 1, wherein The holding member is embedded in a state of being in contact with the object.

3. The embedding container according to claim 1, wherein The embedding container includes an upper portion defining an upper space based on the holding member and a lower portion defining a lower space.

4. The embedding container according to claim 3, wherein The upper portion has an open shape open to the outside.

5. The embedding container according to claim 3, wherein The lower portion has a hole that can be connected to the outside of the embedding container.

6. The embedding container according to claim 1, wherein The holding member includes a first porous member.

7. The embedding container according to claim 6, wherein A second porous member is further provided at a position facing the first porous member with the object interposed therebetween.

8. The embedding container according to claim 6, wherein The thickness of the first porous member in a direction perpendicular to a mounting surface on which the embedding container is mounted is 100 μm or less.

9. The embedding container according to claim 6, wherein The embedding container includes an upper portion defining an upper space and a lower portion defining a lower space based on the holding member. The circumferential length of the lower portion is different from the circumferential length of the upper portion, As a result, a step is formed at the boundary between the upper portion and the lower portion, and the first porous member is arranged on the step.

10. The embedding container according to claim 3, wherein The lower space has an inner diameter of the embedding container that gradually increases as it approaches the upper portion in a direction perpendicular to the installation surface on which the embedding container is installed.

11. The embedding container according to claim 3, wherein The upper space has an inner diameter of the embedding container that gradually decreases as it approaches the lower portion in a direction perpendicular to the installation surface on which the embedding container is installed.

12. The embedding container according to claim 3, wherein The height of the upper portion is not less than 3 mm and not more than 20 mm. The height of the lower portion is not less than 0.2 mm and not more than 3 mm. The inner diameter of the embedding container is not less than 0.5 mm and not more than 6 mm.

13. The embedding container according to claim 3, wherein The plurality of upper spaces are aggregated and configured in a planar shape.

14. The embedding container according to claim 13, wherein The upper spaces that are aggregated and arranged in the planar shape form a honeycomb structure.

15. The embedding container according to claim 1, wherein Made of at least one of plastic, paper, wood and fiber.

16. The embedding container according to claim 1, wherein The object is at least one of a cell, a cell aggregate, a biological tissue, an organism, a fungus, and an algae.

17. The embedding container according to claim 1, wherein further comprising an outer peripheral portion surrounding the outer periphery of the embedding container, The outer peripheral portion has a surface that is not perpendicular to the direction in which the embedding container is cut.

18. The embedding container according to claim 17, wherein The outer peripheral portion includes an intermediate portion extending in the radial direction of the embedding container between the outer peripheral portion and the embedding container. The intermediate portion is a region that is 3 mm or more with respect to the direction in which the embedding container is cut.

19. The embedding container according to claim 6, wherein The embedding container is sliced ​​together with the object to produce a first small piece including the embedding container and the first porous member and a second small piece including the embedding container and the object.

20. An embedding method, characterized in that: Include: a placement step of placing the object in the embedding container according to any one of claims 1 to 19; and An embedding step of embedding the object together with the embedding container.

21. The embedding method according to claim 20, characterized in that: A pretreatment step is further provided after the placement step and before the embedding step. In the pretreatment step, a treatment liquid is introduced into the embedding container with the object placed therein to pretreat the object.

22. The embedding method according to claim 21, characterized in that: After the pretreatment step and before the embedding step, an embedding step is further provided. In the embedding step, an embedding agent is introduced into the embedding container while the object is arranged in the embedding container, thereby embedding the object in the embedding container.

23. An embedding device, characterized in that: include: a moving portion that moves the embedding container according to any one of claims 1 to 19; as well as An introduction portion introduces a treatment liquid and / or an embedding medium into the embedding container.

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