Embedding container and embedding method

Abstract

The present invention provides an embedding container capable of providing an additional approach to methods for embedding cells and tissues. Provided is an embedding container having a placement area where an object to be embedded by an embedding agent can be placed, the embedding container comprising: a sidewall disposed so as to surround the placement area; and a guide part disposed between the placement area and the sidewall to guide the object to the placement area.

Description

Embedding container and embedding method

[0001] The present disclosure relates to an embedding container and an embedding method.

[0002] In cell biology and medical research, specimens are prepared by fixing cells, tissues, etc. with formalin and embedding them in an embedding medium such as paraffin. For example, Patent Document 1 discloses a "tissue cassette assembly for embedding cell blocks." Further techniques for embedding cells and tissues are desired.

[0003] Patent No. 5204810

[0004] The embedding container according to the embodiment is an embedding container having a mounting area where an object to be embedded in an embedding agent can be placed, and is equipped with a side wall arranged to surround the mounting area, and a guide section arranged between the mounting area and the side wall to guide the object to the mounting area.

[0005] FIG. 1 is a conceptual diagram showing the configuration of an embedding container according to an embodiment. FIG. 2 is a block diagram showing a specimen preparation device that executes a specimen preparation process for preparing a specimen from an object accommodated in the embedding container shown in FIG. 1. FIG. 3 is a flowchart showing the specimen preparation process for preparing a specimen from an object accommodated in the embedding container shown in FIG. 1. FIG. 4 is a diagram showing the process of S102 shown in FIG. 3. FIG. 5 is a diagram showing the process of S103 shown in FIG. 3. FIG. 6 is a diagram showing the process of S104 shown in FIG. 3. FIG. 7 is a diagram showing the process of S109 shown in FIG. 3. (a) is a diagram (part 1) showing the thin-sectioning process using a microtome, and (b) is a diagram (part 2) showing the thin-sectioning process using a microtome. FIG. 9 is a conceptual diagram showing the configuration of an embedding container according to a first modified example. FIG. 10 is a conceptual diagram showing the configuration of an embedding container according to a second modified example. FIG. 11 is a conceptual diagram showing the configuration of an embedding container according to a third modified example. FIG. 12 is a conceptual diagram showing the configuration of an embedding container according to a fourth modified example. 13 is a diagram (part 1) showing a specimen preparation process according to a modified example, where (a) shows the process corresponding to step S101 shown in FIG. 3 , (b) shows the process corresponding to steps S102 and S103 shown in FIG. 3 , (c) shows the process of attaching an embedding container to tape, (d) shows the process corresponding to step S104 shown in FIG. 3 , and (e) shows the process of placing the embedding container on a glass slide. FIG. 14 is a diagram (part 2) showing a specimen preparation process according to a modified example, where (a) shows the process corresponding to step S105 shown in FIG. 3 , and (b) shows the process of peeling the tape from the glass slide. (c) shows the process corresponding to steps S106 to S108 shown in FIG. 3 , and (d) shows the process corresponding to step S109 shown in FIG. 3 .

[0006] The embedding container according to the present disclosure will be described below with reference to the drawings. However, it should be noted that the technical scope of the present disclosure is not limited to the embodiments, but extends to the inventions described in the claims and their equivalents.

[0007] 1 is a conceptual diagram showing the configuration of an embedding container according to an embodiment. The embedding container 1 includes a housing 10 and a filter 11, and an object 100 can be placed in a mounting area 12 surrounded by the filter 11. The filter 11 functions as a guide that guides the object 100 to the mounting area 12. The object 100 placed in the mounting area 12 is embedded in an embedding agent (not shown). The embedding container 1 is cut together with the object 100 placed in the mounting area 12 while it is embedded in the embedding agent, thereby creating a specimen of the object 100 that can be observed using a microscope or the like.

[0008] The housing 10 is formed from at least one of plastic, paper, wood, and fiber, and includes a bottom 13 and sidewalls 14. The housing 10 may have a shape such as, but not limited to, a hollow cylinder, a hollow polygonal prism, a hollow truncated cone, or a hollow truncated polygonal pyramid. The center of the bottom 13 of the housing 10 is a mounting area 12 on which the object 100 is placed, and a through-hole 15 into which the filter 11 can be inserted is formed so as to surround the mounting area 12.

[0009] The housing 10 is made of at least one of plastic, paper, wood, and fiber, which facilitates cutting of the embedding container 1 when preparing a specimen containing the object 100. The plastic may be a thermoplastic resin or a thermosetting resin. For example, the plastic forming the housing 10 may include, but is not limited to, 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). The housing 10 is preferably made of an optically transparent material. By forming the housing 10 from an optically transparent material, the object 100 placed in the placement area 12 can be observed.

[0010] When the housing 10 is made of plastic, the housing 10 is easy to process and may be manufactured by any method such as compression molding, injection molding, extrusion molding, etc. When the housing 10 is made of wood, the housing 10 may be manufactured by any method such as thermoforming, etc. When the housing 10 is made of paper or fiber, the housing 10 may be manufactured by any method such as lamination.

[0011] The filter 11 is a porous member having a flat plate shape that allows the chemical solution in which the object 100 is immersed to pass through, and is positioned so that one end is inserted into the through-hole 15. The filter 11 is fixed to the bottom surface 13 of the housing 10 by disposing a coating material bondable to the bottom surface 13 of the housing 10 on its outer edge. The filter 11 may be a mesh filter. Any material may be used for the filter 11, as long as it is not corroded by the chemical solution in which the object 100 is immersed and can withstand the pressure associated with the introduction and discharge of the chemical solution in which the object 100 is immersed. The filter 11 is formed from at least one of plastic, paper, wood, and fiber. For example, the filter 11 is formed from a plastic such as polytetrafluoroethylene (PFTE), polyimide (PI), or polyethersulfone (PES). The filter 11 may also be formed from fiber such as a nonwoven fabric.

[0012] The filter 11 may be coated on the surface facing the object 100 so as to have affinity for the object 100. The coating material for coating the filter 11 may be, but is not limited to, poly-L-lysine, poly-D-lysine, an extracellular matrix (laminin, fibronectin, collagen, or the like), or proteoglycan.

[0013] The thickness of the filter 11 may be 20 μm or more, preferably 200 μm or less, and more preferably 100 μm or less. When the thickness of the filter 11 is 100 μm or more and 200 μm or less, the filter 11 is easy to manufacture and handle. When the thickness of the filter 11 is 20 μm or more and less than 100 μm, less pressure is required to communicate with the chemical solution in which the object 100 is immersed, making it easier to handle solutions such as embedding agents.

[0014] The object 100 is a living organism from which a specimen is to be prepared. The living organism may be an organic living organism. For example, the living organism is a cell. For example, the cell is an animal cell or a plant cell. For example, the cell is a living cell or a dead cell.

[0015] The organism may be an organism other than a cell. As an example, the organism includes at least one of a single cell, a spheroid (aggregate of cells), a biological tissue, an organoid (aggregate of cells), an organism (such as a microorganism), a fungus, and an algae. The organism may include an organelle within a cell. The size of the organism may be, but is not limited to, a major axis of 0.5 mm or less, 1 mm or less, 1.5 mm or less, 2 mm or less, 2.5 mm or less, or 3 mm or less. The size of the organism may be a major axis of 100 μm or less, which is difficult to visually confirm, or 500 μm or less, which is the size of a microstructure.

[0016] 2 is a block diagram showing a specimen preparation device 200 that executes a specimen preparation process for preparing a specimen from the object 100 accommodated in the embedding container 1. The specimen preparation device 200 embeds the object 100 accommodated in the embedding container 1 and prepares a specimen from the embedded object 100. The specimen preparation device 200 includes a communication unit 201, a storage unit 202, an input unit 203, an output unit 204, a control unit 205, a movement unit 206, an organism manipulation unit 207, an introduction unit 208, a cutting unit 209, and a specimen preparation unit 210.

[0017] The communication unit 201 is a component that enables the specimen preparation device 200 to communicate with an external device such as a host control device of the specimen preparation device 200, and includes a communication interface circuit. The communication interface circuit is, for example, a communication interface circuit for a wired LAN, a wireless LAN, LTE (Long Term Evolution), or the like. The communication unit 201 supplies data received from the external device to the control unit 205, and transmits data supplied from the control unit 205 to the external device.

[0018] The storage unit 202 includes, for example, at least one of a semiconductor storage device, a magnetic tape device, a magnetic disk device, and an optical disk device. The storage unit 202 stores an operating system program, a driver program, an application program, data, and the like used in processing by the control unit 205. For example, the storage unit 202 stores a specimen preparation program that causes the control unit 205 to execute a specimen preparation process that embeds the object 100 contained in the embedding container 1 and prepares a specimen from the embedded object 100. The storage unit 202 also stores various information used in the specimen preparation process. The specimen preparation program may be installed into the storage unit 202 from a computer-readable portable storage medium such as a CD-ROM or a DVD-ROM using a known setup program.

[0019] The input unit 203 may be any device capable of inputting data, such as a touch panel or a keyboard. An operator using the specimen preparation device 200 can input letters, numbers, symbols, and the like using the input unit 203. When operated by the operator, the input unit 203 generates a signal corresponding to the operation. The generated signal is then supplied to the control unit 205 as an instruction from the operator.

[0020] The output unit 204 may be any device capable of displaying video, images, etc., such as a liquid crystal display or an organic EL display. The output unit 204 displays video corresponding to video data supplied from the control unit 205, images corresponding to image data, etc. The output unit 204 may also be an output device that prints video, images, text, etc. on a display medium such as paper.

[0021] The control unit 205 includes one or more processors and their peripheral circuits. The control unit 205 is, for example, a CPU, and comprehensively controls the overall operation of the specimen preparation device 200. The control unit 205 executes processing based on programs (such as driver programs, operating system programs, and application programs) stored in the storage unit 202. The control unit 205 can also execute multiple programs (such as application programs) in parallel.

[0022] The moving unit 206 is controlled by the control unit 205 in accordance with an operator's instruction input to the input unit 203, and includes an actuator that moves the embedding container 1. The moving unit 206 places the embedding container 1 inside the specimen preparation device 200 from outside the specimen preparation device 200, and also removes the embedding container 1 placed inside the specimen preparation device 200 and moves it to an embedding dish disposed in the specimen preparation unit 210. The operations of the moving unit 206 are not limited to these, and the moving unit 206 can move the embedding container 1 from any location to any location depending on the purpose.

[0023] The organism manipulation unit 207 is controlled by the control unit 205 in accordance with instructions from an operator input to the input unit 203, and is equipped with a manipulator having a nozzle that manipulates the object 100 contained in the embedding container 1. The organism manipulation unit 207 further includes a pump that supplies and sucks gas into the nozzle flow path, a support unit that moves the nozzle, and the like.

[0024] The introduction unit 208 is controlled by the control unit 205 in response to instructions from an operator input to the input unit 203, and includes a syringe for introducing a liquid into the embedding container 1 and a manifold for accommodating the embedding container 1. The introduction unit 208 introduces a liquid 160 into the embedding container 1 and discharges the liquid 160 from the embedding container 1. For example, the introduction unit 208 introduces a treatment liquid for treating the object 100 into the embedding container 1 and introduces an embedding agent for embedding the object 100 into the embedding container 1.

[0025] The cutting unit 209 is equipped with a cutting tool, and is controlled by the control unit 205 in accordance with instructions from the operator input to the input unit 203, to cut the embedded object 100 with the cutting tool to create a specimen.

[0026] The specimen preparation section 210 performs extension, deparaffinization, staining, dehydration and clearing, and sealing processes on the specimen prepared in the cutting section 209, thereby making the specimen prepared in the cutting section 209 in a state that can be observed by microscope observation, etc.

[0027] Fig. 3 is a flowchart showing a specimen preparation process for preparing a specimen from the object 100 accommodated in the embedding container 1. Fig. 4 is a diagram showing the process of S102 shown in Fig. 3, Fig. 5 is a diagram showing the process of S103 shown in Fig. 3, Fig. 6 is a diagram showing the process of S104 shown in Fig. 3, and Fig. 7 is a diagram showing the process of S109 shown in Fig. 3. The specimen preparation process shown in Fig. 3 is executed mainly by the control unit 205 in cooperation with each element of the specimen preparation device 200, based on a control program stored in advance in the storage unit 202. Note that the specimen preparation process shown in Fig. 3 is one aspect of the embedding method according to the embodiment.

[0028] First, the control unit 205 places the object 100, which has settled in a solvent liquid that dissolves the object 100, in the mounting area 12 of the embedding container 1 via the moving unit 206 and the organism manipulation unit 207 (S101). The control unit 205 forms a bubble at the tip of the nozzle of the organism manipulation unit 207, and brings the object 100 into contact with the gas-liquid interface between the formed bubble and the solvent liquid, thereby recovering the object 100 from the solvent liquid. The control unit 205 may also recover the object 100 by supplying or exhausting a fluid containing a liquid or gas to or from the flow path of the nozzle of the organism manipulation unit 207. The organism manipulation unit 207 may also be a dispenser such as a pipette. Next, the control unit 205 places the recovered object 100 in the mounting area 12 of the embedding container 1. The control unit 205 places the object 100 in the mounting area 12 surrounded by the filter 11. Next, the control unit 205 moves the embedding container 1 into the manifold 110 of the introduction unit 208, with the object 100 placed in the placement area 12. The control unit 205 moves the embedding container 1 via the movement unit 206.

[0029] Next, the control unit 205 performs a chemical pretreatment on the object 100 using various treatment liquids (S102). The control unit 205 introduces various treatment liquids 111 into the embedding container 1 housed in the manifold 110 via syringes to perform pretreatment on the object 100 placed in the embedding container 1. The treatment liquids 111 used in the pretreatment of the object 100 include a fixative, a dehydrating liquid, and a degreasing liquid. The fixative used in the pretreatment may be any of formalin, glutaraldehyde, Bouin's solution, Carnoy's solution, and methanol. The dehydrating liquid used in the pretreatment may be an alcohol such as ethanol, and may be ethanol of any concentration, such as absolute ethanol, 90% ethanol, 80% ethanol, or 70% ethanol. The degreasing liquid used in the pretreatment may be xylene.

[0030] Next, the control unit 205 embeds the object 100 together with the embedding container 1 (S103). The control unit 205 moves the embedding container 1, with the object 100 preprocessed in S102 placed thereon, to the embedding dish 120 arranged in the sampling unit 210. The control unit 205 fills the embedding container 1 contained in the embedding dish 120 with an embedding medium 121 to embed the object 100 placed in the embedding container 1. The embedding medium 121 is, for example, paraffin. The control unit 205 fills the embedding container 1 with liquefied paraffin heated to approximately 60°C until the object 100 is fully immersed, and leaves the embedding container 1 filled with paraffin until the temperature of the heated paraffin drops and the paraffin solidifies. Note that the embedding medium 121 may be celloidin instead of paraffin.

[0031] Next, the control unit 205 thinly slices the object 100 embedded together with the embedding container 1 in the process of S103 to prepare a specimen (S104). The control unit 205 moves the object 100 embedded together with the embedding container 1 from the embedding dish 120 via the moving unit 206. The control unit 205 inverts and positions the embedding container 1 so that the filter 11 is positioned above the object 100. The control unit 205 cuts the inverted embedded object from above using the cutting tool 125 to prepare a thin film including a cross section of the object 100 as a specimen.

[0032] Next, the control unit 205 stretches the specimen prepared in the process of S104 (S105). The control unit 205 moves the specimen prepared in the process of S104 to a water bath filled with water using the moving unit 206, and floats the specimen in the water filled in the water bath, thereby stretching the specimen. The stretched specimen is then moved onto a slide glass by the moving unit 206 and left to dry.

[0033] Next, the control unit 205 deparaffinizes the specimen stretched in the process of S105 (S106). The control unit 205 moves the specimen together with the slide glass using the moving unit 206, and deparaffinizes the specimen by sequentially immersing the specimen in doses filled with xylene, 100% ethanol, and 70% ethanol, respectively.

[0034] Next, the control unit 205 stains the specimen deparaffinized in the process of S106 with hematoxylin-eosin staining (HE staining) (S107). The control unit 205 moves the specimen together with the slide glass using the moving unit 206 and sequentially immerses the specimen in doses filled with hematoxylin solution and eosin solution, respectively, to stain the specimen. Note that the control unit 205 may stain the specimen with a known HE staining method such as Klüver-Barrera staining (KB staining), Fontana-Masson staining, Giemsa staining, Gram staining, or toluidine blue staining.

[0035] Next, the specimen stained in the process of S107 is dehydrated and cleared (S108). The control unit 205 moves the specimen together with the slide glass using the moving unit 206, and dehydrates and clears the specimen by sequentially immersing the specimen in doses filled with 70% ethanol, 100% ethanol, and xylene, respectively.

[0036] The control unit 205 then mounts the specimen 131 dehydrated and cleared in the process of S108 on the glass slide 130 (S109). The specimen 131 is mounted by placing a mounting medium on the glass slide 130 on which the dehydrated and cleared specimen is placed, and then placing a cover glass 132 on the mounting medium placed on the glass slide 130. The mounting medium may contain glycerin, or a commercially available mounting medium may be used. The periphery of the cover glass 132 may be sealed with commercially available nail polish or the like.

[0037] The embedding container 1 prevents the object 100 from being placed at the edge of the housing 10 by making the filter 11 function as a guide that guides the object 100 to the mounting area 12 located at the center of the housing 10. Preventing the object 100 from being placed at the edge of the housing 10 prevents the object 100 from interfering with the housing 10 and being damaged when the embedded object 100 is sliced. Furthermore, the embedding container 1 allows the object 100 to be placed in the mounting area 12 surrounded by the filter 11, which limits movement of the object 100 when a chemical solution is passed through it.

[0038] Furthermore, by using the filter 11 as a guide, the embedding container 1 can guide the object 100 to the mounting area 12 located at the center of the housing 10 without having to install any additional components that function as a guide to guide the object 100.

[0039] Furthermore, in the embedding container 1, the filter 11 has a flat plate-like shape, which makes it easy to handle and place the filter 11, and also reduces the manufacturing cost of the filter 11 compared to when a filter with a complex shape is used.

[0040] The specimen preparation device 200 prepared the specimen by moving the embedding container 1 to the embedding dish 120 arranged in the specimen preparation section 210. However, the specimen preparation device according to the embodiment may be configured to perform pre-processing and embedding of the object 100 without moving the embedding container 1 to the embedding dish 120, and to perform thin-sectioning with the embedding container 1 directly attached to a microtome.

[0041] FIG. 8(a) is a diagram (part 1) showing the thin-sectioning process using a microtome, and FIG. 8(b) is a diagram (part 2) showing the thin-sectioning process using a microtome.

[0042] In the specimen preparation device according to the modified example, after pre-processing and embedding of the object 100, the object is attached directly to a microtome and subjected to thin-sectioning. In the specimen preparation device according to the modified example, the embedding process may be performed so that the embedding medium 123 fills the area surrounded by the filter 11, or so that the embedding medium 123 fills the entire interior of the housing 10. In the specimen preparation device according to the modified example, the embedding process is performed, and after the embedding medium 123 has been filled inside the housing 10, the bottom surface 13 of the housing 10 is cut with a cutting tool 125 while the bottom surface 13 of the housing 10 is protruding from the microtome, thereby performing thin-sectioning.

[0043] Furthermore, in the embedding container 1, the filter 11 is fixed to the bottom surface 13 of the housing 10 by disposing a coating material on its outer edge; however, the filter 11 may be fixed to the housing 10 by other methods. For example, the filter 11 may be fixed to the housing 10 by disposing a fixing member between the side surface 14 of the housing 10 and the filter 11. When the filter 11 is fixed by a fixing member disposed between the side surface 14 and the filter 11, it is preferable that the fixing member be removed after the embedding medium is filled and before the embedding medium solidifies. Note that in the specimen preparation device according to the modified example, the embedding container 1 is directly attached to the microtome without being moved to the embedding dish 120 to perform the thin-sectioning process; however, instead, the pre-processing, embedding process, and thin-sectioning process of the object 100 may be performed using a single microtome.

[0044] 9 is a conceptual diagram showing the configuration of an embedding container according to a first modified example. The embedding container 2 includes a housing 20, a filter 21, and a guide member 22. An object 100 can be placed in a mounting area 23 surrounded by a guide portion formed by the filter 21 and the guide member 22. The filter 21 and the guide member 22 function as a guide portion that guides the object 100 to the mounting area 23. The object 100 placed in the mounting area 23 is embedded in an embedding agent (not shown). The embedding container 2 is cut together with the object 100 placed in the mounting area 23 while it is embedded in the embedding agent, thereby producing a specimen of the object 100 that can be observed using a microscope or the like.

[0045] Similar to the housing 10, the housing 20 is made of at least one of plastic, paper, wood, and fiber and has a cylindrical shape. The housing 20 has side surfaces that are arranged to surround the mounting area 23. A locking portion for locking the filter 21 is formed on the inner wall of the housing 20. Similar to the filter 11, the filter 21 is a porous member having a flat plate shape that is permeable to the chemical solution in which the object 100 is immersed, and is locked to the housing 20 by the locking portion formed on the inner wall of the housing 20.

[0046] The guide member 22 is formed from an embedding agent and is disposed on the filter 21 between the mounting area 23 and the housing 20 so as to surround the mounting area 23. The guide member 22 is formed from an embedding agent, but may be formed from a dissolvable material other than an embedding agent. The guide member 22 has an inclined surface 24 that inclines so as to approach the inner wall of the housing 20 as it becomes more distant from the mounting area 23.

[0047] The specimen preparation process for preparing a specimen from the object 100 accommodated in the embedding container 2 is executed in the same manner as the specimen preparation process shown in Fig. 3, except for the process of embedding the object 100 shown in S103. When preparing a specimen from the object 100 accommodated in the embedding container 2, the housing 20 is heated to dissolve the guide member 22 and embed the object 100, and an embedding medium is added and filled into the embedding container 2 until the object 100 is sufficiently immersed.

[0048] The embedding container 2 has a guide member 22 formed of an embedding agent placed on the filter 21, so by using the guide member 22 as the embedding agent for embedding the object 100, the embedding process is easier than with the embedding container 1.

[0049] Furthermore, by making the placement area 23 on which the object 100 is placed the central area of ​​the filter 21, the processing liquid used for pre-processing can be discharged to the outside through the placement area 23, and pre-processing can be performed without discharging the embedding material that forms the guide member 22 to the outside.

[0050] Furthermore, since the guide member 22 of the embedding container 2 is formed from an embedding agent, the guide member 22 can be formed in a shape that guides the target object 100 to the mounting area 23. Specifically, the embedding container 2 includes an inclined surface 24 that inclines so that the guide member 22 approaches the inner wall of the housing 20 as it moves away from the mounting area 23, thereby enabling the target object 100 to be guided to the mounting area 23.

[0051] 10 is a conceptual diagram showing the configuration of an embedding container according to a second modification. The embedding container 3 includes a housing 30, a filter 31, and a guide member 32. An object 100 can be placed in a placement area 33 surrounded by a guide portion formed by the filter 31 and the guide member 32. The filter 31 and the guide member 32 function as a guide portion that guides the object 100 to the placement area 33. The object 100 placed in the placement area 33 is embedded in an embedding agent (not shown). The embedding container 3 is cut together with the object 100 placed in the placement area 33 while it is embedded in the embedding agent, thereby producing a specimen of the object 100 that can be observed using a microscope or the like.

[0052] Similar to the housing 10, the housing 30 is made of at least one of plastic, paper, wood, and fiber, and includes a bottom 35 and sidewalls 36. The bottom 35 includes a light-transmitting portion 37 located in the center, with a through-hole 38 formed around the light-transmitting portion 37 and locked to the sidewalls 36 by a locking portion (not shown). A mounting area 33 on which the object 100 is placed is located in the light-transmitting portion 37. Locking portions for locking the filter 31 are formed on the bottom 35 and sidewalls 36. Similar to the filter 11, the filter 31 is a porous member having a flat plate shape that is permeable to the chemical solution in which the object 100 is immersed. The filter 31 is locked to the housing 30 by the locking portions formed on the bottom 35 and sidewalls 36, thereby covering the through-hole 38.

[0053] The guide member 32 is formed from an embedding agent and is disposed on the filter 31 between the mounting area 33 and the side wall 36 so as to surround the mounting area 33. The guide member 32 is formed from an embedding agent, but may be formed from a dissolvable material other than an embedding agent. The guide member 32 has an inclined surface 34 that slopes toward the side wall 36 as it moves away from the mounting area 33.

[0054] The specimen preparation process for preparing a specimen from the object 100 accommodated in the embedding container 3 is executed in the same manner as the specimen preparation process shown in Fig. 3 , except that a process for removing the guide member 32 is added between the process shown in S101 and the process shown in S102. In the specimen preparation process for preparing a specimen from the object 100 accommodated in the embedding container 3, after the object 100 is placed in the process shown in S101, the housing 30 is heated to dissolve the guide member 32. The dissolved guide member 32 is removed through the filter 31 by being sucked by a suction device (not shown).

[0055] The embedding container 3 has a mounting area 33 on which the object 100 is placed, which is arranged in a light-transmitting section 37, so that the object 100 placed in the mounting area 33 can be seen through the light-transmitting section 37, making it easy to slice the object 100 thinly.

[0056] 11 is a conceptual diagram showing the configuration of an embedding container according to a third modification. The embedding container 4 includes a housing 40, a filter 41, and a support member 42. The object 100 can be placed in a placement area 43 surrounded by a guide formed by the filter 41 and the support member 42. The filter 41 and the support member 42 function as a guide that guides the object 100 to the placement area 43. The object 100 placed in the placement area 43 is embedded in an embedding agent (not shown). The embedding container 4 is cut together with the object 100 placed in the placement area 43 while it is embedded in the embedding agent, thereby creating a specimen of the object 100 that can be observed using a microscope or the like.

[0057] Similar to the housing 10, the housing 40 is made of at least one of plastic, paper, wood, and fiber, and includes a bottom 45 and sidewalls 46. The bottom 45 includes a light-transmitting portion 47 located in the center, with a through-hole 48 formed around the light-transmitting portion 47 and secured to the sidewalls 46 by a locking portion (not shown). A mounting area 43 on which the object 100 is placed is located in the light-transmitting portion 47. Locking portions for locking the filter 41 and the support member 42 are formed on the bottom 45 and sidewalls 46. Similar to the filter 11, the filter 31 is a porous member having a flat plate shape that is permeable to the chemical solution in which the object 100 is immersed. The filter 31 is secured to the housing 30 by the locking portions formed on the bottom 45 and sidewalls 46, thereby forming an inclined surface 44.

[0058] The support member 42 is a porous member made of a synthetic resin such as polyurethane, and is disposed below the filter 41 so as to surround the mounting area 43 between the mounting area 43 and the side wall 46. The support member 42 has a plurality of holes formed therein through which the liquid and embedding medium that have passed through the filter 41 pass. The support member 42 supports the filter 41 so as to form an inclined surface 44 that slopes toward the side wall 46 as it moves away from the mounting area 43.

[0059] The specimen preparation process for preparing a specimen from the object 100 accommodated in the embedding container 4 is carried out in the same manner as the specimen preparation process shown in FIG.

[0060] The embedding container 4 is arranged below the filter 44 to support the filter 44, and is provided with a support member 42 that is permeable to liquid that has passed through the filter 44, so that the filter 44 can be arranged to guide the object 100 to the placement area 43.

[0061] Specifically, the embedding container 4 is positioned at an angle so that the filter 44 approaches the side wall 46 of the housing 40 as it moves away from the mounting area 43, thereby guiding the object 100 to the mounting area 43.

[0062] Furthermore, by arranging the support member 42 in the embedding container 4 so as to cover the through-hole 48 , the liquid that has passed through the filter 44 can be discharged to the outside of the embedding container 4 via the support member 42 .

[0063] 12 is a conceptual diagram showing the configuration of an embedding container according to a fourth modification. The embedding container 5 includes a housing 50 and a filter 51. The filter 51 has a recess 52 formed in it, and a mounting area 53 on the bottom surface of the recess 52 allows the object 100 to be mounted thereon. The filter 51 functions as a guide that guides the object 100 to the mounting area 53. The object 100 mounted in the mounting area 53 is embedded in an embedding agent (not shown). The embedding container 5 is cut together with the object 100 mounted in the mounting area 53 while it is embedded in the embedding agent, thereby producing a specimen of the object 100 that can be observed under a microscope or the like.

[0064] Similar to the housing 10, the housing 50 is made of at least one of plastic, paper, wood, and fiber and has a cylindrical shape. The housing 50 has side surfaces arranged to surround the mounting area 53. A locking portion for locking the filter 51 is formed on the inner wall of the housing 50.

[0065] The filter 51, like the filter 11, is a porous member and is fastened to the housing 50 by fastening portions formed on the inner wall of the housing 20. The filter 51 has a cylindrical central portion 54 and a cylindrical peripheral portion 55 arranged to surround the central portion 54. The height of the central portion 54 is lower than the height of the peripheral portion 55, and a recess 52 is formed above the central portion 54. The surface of the central portion 54 is a mounting area 53 on which the object 100 can be placed. The thickness of the central portion 54 may be 10 μm or more and 20 μm or less.

[0066] By placing a single filter 51 inside the housing 50, the embedding container 5 can form a guide section that guides the object 100 to the mounting area 53, thereby achieving the function of guiding the object 100 to the mounting area 12 located at the center of the housing 10 using a simple manufacturing method.

[0067] In the embedding containers 1 to 5, the guide portion that guides the object 100 to the placement area is formed at least in part by a filter, but in the embedding container of the embodiment, the guide portion that guides the object 100 to the placement area does not have to be formed by a filter.

[0068] In addition, in the specimen preparation process shown in Figure 3, the specimen is prepared by sequentially executing the processes of S101 to S109, but in the specimen preparation process according to the embodiment, the processes of S104 to S109 may be performed by attaching the embedded embedding container to tape.

[0069] FIG. 13 is a diagram (part 1) showing a specimen preparation process according to a modified example, and FIG. 14 is a diagram (part 2) showing a specimen preparation process according to a modified example. FIG. 13(a) shows a process corresponding to step S101 shown in FIG. 3 , FIG. 13(b) shows a process corresponding to steps S102 and S103 shown in FIG. 3 , and FIG. 13(c) shows a process for attaching an embedding container to tape. FIG. 13(d) shows a process corresponding to step S104 shown in FIG. 3 , and FIG. 13(e) shows a process for placing an embedding container on a glass slide. FIG. 14(a) shows a process corresponding to step S105 shown in FIG. 3 , and FIG. 14(b) shows a process for peeling the tape from the glass slide. FIG. 14(c) shows a process corresponding to steps S106 to S108 shown in FIG. 3 , and FIG. 14(d) shows a process corresponding to step S109 shown in FIG. 3 .

[0070] First, the object is placed in an embedding container 6 shown in Fig. 13(a). Next, as shown in Fig. 13(b), the embedding container 6 with the object placed therein is placed in a microtome 60 capable of performing pre-processing and embedding. Next, the object placed in the embedding container 6 placed in the microtome 60 is subjected to pre-processing and then embedded. Next, after the object placed in the embedding container 6 has been embedded, tape 61 is attached to the embedding container 6 as shown in Fig. 13(c).

[0071] Next, as shown in Figure 13(d), the embedding material 62 adhering to the tape 61 peeled off from the embedding container 6 is cut to prepare a specimen. Next, as shown in Figure 13(e), the tape 61 is attached to a slide glass 63. By attaching the tape 61 to the slide glass 63, the specimen 64 attached to the tape 61 is placed on the slide glass 63. When the tape 61 is attached to the slide glass 63, a liquid is applied to the area where the specimen 64 will be placed. By applying the liquid to the area where the specimen 64 will be placed, the adhesive strength between the slide glass 63 and the specimen 64 is improved.

[0072] 14(a), the slide glass 63 with the tape 61 attached thereto is stored for a predetermined time, such as 12 hours, at a temperature higher than room temperature, such as 47° C. By storing the slide glass 63 at a temperature higher than room temperature for the predetermined time, the specimen 64 attached to the tape 61 stretches.

[0073] 14(b), the tape 61 attached to the slide glass 63 is peeled off from the slide glass 63. When the tape 61 is peeled off from the slide glass 63, the adhesive force between the slide glass 63 and the specimen 64 is stronger than the adhesive force between the tape 61 and the specimen 64, so the specimen 64 remains disposed on the slide glass 63.

[0074] Next, as shown in Fig. 14(c), the specimen 64 placed on the slide glass 63 is stained, dehydrated, and cleared. Then, as shown in Fig. 14(d), the stained, dehydrated, and cleared specimen 64 is sealed with a cover glass 55, and the specimen preparation process is completed.

[0075] 1 to 6 Embedding container 10, 20, 30, 40, 50 Housing 11, 21, 31, 41, 51 Filter 12, 23, 33, 43, 53 Placement area 13, 35, 45 Bottom 14, 36, 46 Side wall 100 Object

Claims

1. An embedding container having a placement area where an object to be embedded in an embedding agent can be placed, the embedding container comprising: a side wall arranged to surround the placement area; and a guide section arranged between the placement area and the side wall, which guides the object to the placement area.

2. The embedding container according to claim 1, wherein at least a part of the guide portion is formed by a filter that is permeable to the chemical solution in which the object is immersed.

3. The embedding container according to claim 2, wherein the guide portion is formed by the filter and the embedding medium.

4. The embedding container according to claim 3, wherein the filter is supported by the side walls, and the mounting area is a central area of ​​the filter.

5. An embedding container as described in claim 3, further comprising a bottom having a light-transmitting portion located in the center and a through-hole formed around the light-transmitting portion, the filter being disposed in the through-hole, and the mounting area being disposed in the light-transmitting portion.

6. An embedding container according to claim 3, wherein the embedding medium has an inclined surface that slopes toward the side wall as it moves away from the placement area.

7. The embedding container according to claim 2, further comprising a support member disposed below the filter so as to support the filter, the support member being permeable to the liquid that has passed through the filter.

8. The embedding container according to claim 7, further comprising a bottom having a light-transmitting portion located in the center and a through hole formed around the light-transmitting portion, and the support member is positioned so as to cover the through hole.

9. The embedding container according to claim 7, wherein the filter is disposed at an angle so as to approach the side wall as it moves away from the placement area.

10. An embedding method for embedding an object using an embedding container having a mounting area on which an object to be embedded in an embedding agent can be placed, the embedding container comprising: side walls arranged to surround the mounting area; a bottom having a centrally located light-transmitting section and a through-hole formed around the light-transmitting section; and a guide section formed at least in part by a filter permeable to a chemical solution in which the object is immersed, the guide section being arranged to cover the through-hole and guiding the object to the mounting area, the method comprising the steps of placing the object in the mounting area, performing a pre-processing step of immersing the object placed in the mounting area in a liquid, and embedding the object after the pre-processing step in an embedding agent, wherein the liquid in which the object has been immersed during the pre-processing step is discharged via the filter.

11. The embedding method according to claim 10, wherein the guide portion is formed by the filter placed in the through hole and an embedding agent placed to cover the filter, and the pre-processing is performed after heating the embedding container to dissolve the embedding agent and remove it through the filter.

12. The embedding method according to claim 10, wherein the guide section is formed by the filter and a support member that is positioned below the filter to support the filter and is permeable to the liquid that has passed through the filter, and the liquid in which the object is immersed during the pre-processing is discharged via the filter and the support member.

13. An embedding device comprising: a storage section for storing an embedding container according to any one of claims 1 to 9; and an introduction section for introducing a processing liquid and / or an embedding medium into the embedding container stored in the storage section.

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