JP2013185920A - Method for manufacturing tissue array sheet and manufacturing device thereof - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for manufacturing a tissue array sheet capable of manufacturing a tissue array including multiple cores arranged on a slide, and a manufacturing device thereof.SOLUTION: The method for manufacturing tissue array sheets includes the following steps of: (a) sticking a film to a surface of a tissue block; (b) slicing the surface side of the tissue block with the film and acquiring a tissue piece with the film; (c) sticking a hollow needle into the tissue piece of the tissue piece with the film, boring it, and retaining a small piece of the tissue piece in the interior of the hollow needle; (d) repeating the step (c) and obtaining a laminate formed from a plurality of small pieces in the interior of the hollow needle; (e) heating the laminate and obtaining a fusion product; (f) inserting the fusion product into a hole part of a substrate block; and (g) slicing the substrate block as traversing the laminate.

Description

  The present invention relates to a method and apparatus for manufacturing a tissue array sheet applied to general pathological diagnosis and research. More specifically, the present invention relates to a tissue array sheet manufacturing method and a manufacturing apparatus for manufacturing a tissue array chip used for examination and analysis of living tissue.

  Conventionally, a tissue array chip in which tissue pieces are arranged on a substrate has been used for examination and analysis of a living tissue. The tissue array chip is used for examination of the presence or absence of a diseased tissue, analysis of genes and proteins, screening, and the like by applying a staining solution that specifically stains a test substance.

  This tissue array chip is manufactured as follows (FIG. 14).

  (A) A core is collected by punching a tissue block obtained by blocking a living tissue. (b) The collected cores are inserted into holes provided in an array on the base block to produce a tissue array block. (c) The surface of a base block (that is, a tissue array block) in which a core is inserted into each hole is sliced on the order of several μm to produce a tissue array sheet in which tissue pieces are arranged. (D) A tissue array chip is fabricated by mounting the tissue array sheet on a substrate. A plurality of tissue array chips are manufactured by repeatedly performing the above steps (c) and (d).

  The tissue array block used for manufacturing the tissue array chip is manufactured by an apparatus having a punching mechanism. In Patent Document 1 and Patent Document 2, a punch for a base block and a punch for a tissue block are provided, a hole is formed in the base block with the punch for the base block, and the tissue block is punched with the punch for the tissue block. An apparatus for collecting a core and inserting the collected core into a hole in a base block is disclosed.

Patent Documents 1 and 2 both disclose a method of manufacturing a tissue array in which a hole is formed in a tissue block, a core is extracted (ie, coring is performed), and transplanted to a substrate block that is a recipient block. In producing a tissue array chip using these methods, the following many problems are assumed.
(1) The tissue cannot be observed again at the coring site. For this reason, it may not be approved by the Ethics Committee.
(2) There is a facility that prohibits the organization block from being taken outside. In such a facility, multiple facilities cannot jointly produce an array.
(3) Interesting cases and valuable encouraged tissue blocks are treasures of pathologists, and it is extremely difficult to make holes in tissue blocks.
(4) The extraction of the core is limited to a tissue block having a certain thickness.
(5) When a target lesion, such as a tumor, is flying away, it is difficult for a normal core to make the entire section a lesion.
(6) Since a tissue block having a different thickness is used, a core chip may occur.

  As means for solving the above problems, the present inventor has filed an application for a technique (spiral tissue array) for producing a tissue array block and a tissue array sheet from a sliced tissue piece formed in a roll shape (Patent Document 3). .

  The spiral tissue array is an excellent technique that substantially solves the drawbacks of conventional tissue arrays produced by punching.

  However, in the spiral tissue array, since the sliced tissue pieces are formed in a roll shape, the core diameter increases due to winding. On the other hand, since the tissue array sheet is placed on a slide glass (2.5 cm × 7.5 cm) and observed, it is usually 2 cm × 3 cm in size. Therefore, in the spiral tissue array, the number of tissues placed on one sheet is limited to about several tens.

  Further, when slicing a tissue block, there is a disadvantage that the sliced tissue slices are deformed, resulting in poor workability.

JP 2004-215667 A JP 2006-158394 A JP 2008-28410 A

  The present invention has been made to solve the above-described prior art, and an object of the present invention is to produce a tissue array block that can produce a tissue array sheet without making holes in the tissue block, and to produce a tissue array sheet. It is to provide a method and a manufacturing apparatus thereof.

  Another object of the present invention is to provide a method of manufacturing a tissue array block, a method of manufacturing a tissue array sheet, and a manufacturing apparatus thereof that can produce a small-diameter core and increase the number of cores.

  Still another object of the present invention is to remove only the lesion, so that the target lesion site (region of interest) is comprehensively placed in the laminated / fused tissue piece, and as a result, all or most of the laminated / fused tissue piece is removed. An object of the present invention is to provide a method for manufacturing a tissue array block derived from a lesion site (region of interest), a method for manufacturing a tissue array sheet, and a manufacturing apparatus therefor.

  Still another object of the present invention is to select a tissue site to be adopted as a laminated / fused tissue piece while observing the heterogeneity of the tissue, so that the heterogeneity is highly comprehensive (ie, rich in diversity). An object of the present invention is to provide an array block manufacturing method, a tissue array sheet manufacturing method, and a manufacturing apparatus thereof.

  Still another object of the present invention is to provide a method for manufacturing a tissue array block, a method for manufacturing a tissue array sheet, and a manufacturing apparatus for the same, because an array can be manufactured with sliced tissue pieces, and thus facilitates joint research among multiple facilities. Is to provide.

  Still another object of the present invention is to accurately embed a laminated body hollowed out by a hollow needle through a through hole of a heating base, and to reduce the size of the laminated body to be hollowed out and the size of the through hole of the heating base. As a result, there are provided a method of manufacturing a tissue array block, a method of manufacturing a tissue array sheet, and a manufacturing apparatus thereof capable of manufacturing a tissue array in which a large number of laminated / fused tissue pieces are arranged on a slide. There is.

  Still another object of the present invention is to provide a method for manufacturing a tissue array block, a method for manufacturing a tissue array sheet, and a method for manufacturing the same, which can retain a slice of tissue sliced when slicing the tissue block and have excellent workability. To provide an apparatus.

  In the method for producing a tissue array block of the present invention, a film is attached to the surface of the tissue block, a tissue piece with a film is sliced (sliced) from the tissue block, and the sliced (sliced) tissue piece with a film is obtained. A plurality of tissue pieces collected in the hollow needle are laminated by hollowing out a part of a necessary tissue piece using a hollow needle and repeating this hollowing process, and the laminate is heated and fused to form a fusion ( A layered / fused tissue piece) is inserted in the holes arranged in the base block or the like in the axial direction.

  Moreover, this invention is a tissue array sheet | seat produced by slicing the tissue array block produced by the said process. Tissue blocks used in the manufacture of tissue array blocks include, but are not limited to, paraffin-embedded tissue blocks and frozen tissue blocks. Paraffin includes paraffin that does not contain an additive and paraffin that contains an additive of a synthetic polymer such as polyethylene.

Accordingly, the present invention provides the following.
(Item 1)
(a) attaching a film to the surface of the tissue block;
(b) slicing the surface side of the tissue block to which the film is attached to obtain a tissue piece to which the film is attached;
(c) piercing a hollow needle into the tissue piece of the tissue piece with film, and holding the small piece of the tissue piece inside the hollow needle;
(D) repeating the above step (c) to obtain a laminate comprising a plurality of small pieces inside the hollow needle;
(E) heating the laminate to obtain a fusion;
(F) inserting the fusion into a hole provided in the base block;
(G) slicing the base block across the laminate;
For producing a tissue array sheet comprising:
(Item 2)
(a) attaching a film to the surface of the tissue block;
(b) slicing the surface side of the tissue block to which the film is attached to obtain a tissue piece to which the film is attached;
(c) piercing a hollow needle into the tissue piece of the tissue piece with film, and holding a small piece of the tissue piece inside the hollow needle;
(D) repeating the above step (c) to obtain a laminate comprising a plurality of small pieces inside the hollow needle;
(E) heating the laminate to obtain a fusion;
(F) The fusion product is placed in a through-hole of a pallet arranged in the embedding dish, and a molten paraffin solution is poured into the embedding dish to obtain a tissue array block in which the fusion product and paraffin are integrated. Process,
(G) slicing the tissue array block across the fusion;
For producing a tissue array sheet comprising:
(Item 3)
(a) attaching a film to the surface of the tissue block;
(b) slicing the surface side of the tissue block to which the film is attached to obtain a tissue piece to which the film is attached;
(c) piercing a hollow needle into the tissue piece of the tissue piece with film, and holding the small piece of the tissue piece inside the hollow needle;
(D) repeating the above step (c) to obtain a laminate comprising a plurality of small pieces inside the hollow needle;
(E) heating the laminate to obtain a fusion;
(F) inserting the fusion into a hole provided in the base block;
For producing a tissue array block comprising:
(Item 4)
(a) attaching a film to the surface of the tissue block;
(b) slicing the surface side of the tissue block to which the film is attached to obtain a tissue piece to which the film is attached;
(c) piercing a hollow needle into the tissue piece of the tissue piece with film, and holding a small piece of the tissue piece inside the hollow needle;
(D) repeating the above step (c) to obtain a laminate comprising a plurality of small pieces inside the hollow needle;
(E) heating the laminate to obtain a fusion;
(F) placing the fusion in a through-hole of a pallet arranged in the embedding dish, pouring molten paraffin liquid into the embedding dish, and integrating the fusion and paraffin;
For producing a tissue array block comprising:
(Item 5)
The manufacturing method of the tissue array block in any one of the items 1-4 whose said film is 1 type selected from the group which consists of a polyethylene terephthalate, a polyvinyl chloride, paper, and a polypropylene film.
(Item 6)
A base, and a table placed on the base and movable in the horizontal X-axis and Y-axis directions by operating the moving means;
Elevating means attached to the base and arranged to be elevable;
A tissue punching means for punching out the tissue piece of the tissue piece with a film attached to the lifting means and placed on the table by the lowering of the lifting means;
Means for punching the tissue piece of the tissue piece with a film a plurality of times by the tissue cutting means to obtain a laminate;
A heating device for heating the laminate;
An apparatus for manufacturing a tissue array sheet.
(Item 7)
Item 7. The apparatus according to Item 6, wherein the heating means is disposed on a table.
(Item 8)
The heating means includes a heating base having a hole and excellent heat conductivity, a heater for heating the heating base, a temperature sensor, and a temperature control device having a feedback function. Item 6. The device according to Item 6.
(Item 9)
Item 7. The apparatus according to Item 6, wherein the heating means is a warm air blower that applies warm air to the hollow needle.
(Item 10)
The apparatus according to item 6, further comprising cooling means disposed on the table.
(Item 11)
A film used for producing a tissue array sheet, the film sticking to the surface of the tissue block, and also sticking to the tissue block after slicing the tissue piece with film from the tissue block A film having adhesiveness to the tissue block.
(Item 12)
Item 12. The film according to Item 11, wherein an acrylic pressure-sensitive adhesive is applied to a surface that sticks to the surface of the tissue block.
(Item 13)
Item 12. The film according to Item 11, comprising a substance selected from the group consisting of polyethylene terephthalate, polyvinyl chloride, paper, and polypropylene film as a main component.
(Item 14)
Tissue piece storage device, including:
(A) film;
(B) a tissue piece; and (c) a substrate,
Here, the (b) tissue piece is located between the (a) film and the (c) substrate,
The (b) preservation | save implement in which the surface of this (a) film which contact | connects a tissue piece has adhesiveness with respect to a tissue piece.
(Item 15)
Item 15. The storage device according to Item 14, wherein an acrylic adhesive is applied to the surface of the (a) film in contact with the (b) tissue piece.
(Item 16)
Item 15. The storage device according to item 14, wherein the (a) film is mainly composed of a substance selected from the group consisting of polyethylene terephthalate, polyvinyl chloride, paper, and polypropylene film.
(Item 17)
Item 15. The storage device according to Item 14, wherein (a) the base material contains as a main component a substance selected from the group consisting of polyethylene terephthalate, polyvinyl chloride, paper, and polypropylene film.

  According to the present invention, one tissue array sheet can be produced with the same number of tissues as the number of tissues placed on a tissue array sheet produced by conventional punching.

The front view of the state which affixed the film on the organization block. Explanatory drawing of the state which cuts out the slice with a hollow needle, after slicing the tissue piece with a film from the tissue block which stuck the film. The expanded sectional view which shows the state which has arrange | positioned the laminated body in the hollow cylinder of a hollow needle to a heating apparatus. The perspective view of the state which inserts a fusion | fusion thing in the several hole of a base | substrate block. The expanded sectional view which inserted the fusion | fusion thing in the hole of a base | substrate block. Explanatory drawing which shows the process of slicing the surface of a tissue array block. The schematic front view which fractured | ruptured a part of manufacturing apparatus of a structure | tissue array block. It is a schematic front view of the manufacturing apparatus of a tissue array block, and shows the state where the table is moved. The top view which fractured | ruptured partially the manufacturing apparatus of a structure | tissue array block. The right view which fractured | ruptured partially the manufacturing apparatus of a structure | tissue array block. It is right sectional drawing of the manufacturing apparatus of a tissue array block, Comprising: The state which operated the handle | steering-wheel is shown. Explanatory drawing which shows the state which moved the table. Explanatory drawing which shows the state which arrange | positions the fusion | melting material in a hollow needle in an embedding dish through the through-hole of a pallet. Schematic which shows the manufacturing method of the conventional tissue array chip | tip. Sectional drawing which illustrates the preservation | save tool of this invention. The schematic diagram which shows the usage example of the preservation | save implement of this invention.

  The tissue array block produced in the method for producing a tissue array sheet of the present invention is a tissue array block in which tissue pieces are held in a plurality of holes provided in an array on a base block. ) Affixing a film on the surface of the tissue block, (ii) slicing (slicing) a tissue piece with film from the tissue block, and (iii) using a hollow needle for the sliced (slicing) tissue piece with film And (iv) laminating a plurality of tissue pieces collected in the hollow needle by repeating this hollowing process, and (v) heating and fusing the laminate. It was prepared by this.

  The tissue array sheet of the present invention is sliced from a tissue array block that holds a plurality of laminated / fused tissue pieces in a plurality of holes provided in an array on the base block.

The tissue array block manufacturing method and tissue array sheet of the present invention have the following advantages.
(1) A tissue array sheet can be produced without making holes in the tissue block.
(2) A small-diameter laminated / fused tissue piece can be produced, and the number of laminated / fused tissue pieces that can be held per base block can be increased.
(3) Since it is possible to select only the lesioned part and prepare the laminated / fused tissue piece, the target lesion site (site of interest) can be comprehensively placed in the laminated / fused tissue piece.
(4) Since the tissue site to be adopted as the laminated / fused tissue piece can be selected while observing the heterogeneity of the tissue, the comprehensiveness of the heterogeneity is high.
(5) Since an array can be produced with sliced tissue pieces, collaborative research among multiple facilities is facilitated.
(6) When slicing a tissue block, a slice of the sliced tissue can be shaped and excellent in workability.

  The present invention is described in detail below.

(the film)
The film (22) used for the production of the tissue array sheet of the present invention,
(A) Adhesiveness to a tissue block that adheres to the surface of the tissue block and maintains the state adhered to the tissue block even after slicing the tissue piece with film from the tissue block;
(B) Releasability that allows the hollowed-out tissue piece to be released after hollowing out a part of the tissue piece;
(C) the ability to retain the shape after slicing the tissue piece with film from the tissue block;
(D) Strength that is not cut when a part of a tissue piece is cut out by a hollow needle;
Any adhesive film can be used as long as it is provided with the adhesive property of (a) and the releasability of (b), for example, by applying an adhesive to the film surface to be attached to the tissue block. Achieved by: Examples of such an adhesive include, but are not limited to, acrylic pressure-sensitive adhesives.

  The shape retention ability of (c) and the strength of (d) can be achieved, for example, by producing a film with a material selected from the group consisting of polyethylene terephthalate, polyvinyl chloride, paper, and polypropylene film. When using paper, the surface of the paper may or may not be mirror-finished.

  The strength of the above (d) can be easily achieved by adjusting the thickness of the film. The thickness of the film can be appropriately determined by those skilled in the art. Typically, in the case of a film selected from the group consisting of polyethylene terephthalate, polyvinyl chloride, paper, and polypropylene film, about 20 μm to 200 μm. Is the thickness.

(Method for producing tissue array sheet)
The tissue array sheet of the present invention uses the above-mentioned film, (i) a film is attached to the surface of the tissue block, (ii) a tissue piece with a film is sliced (sliced) from the tissue block, and (iii) the slice A part of the required tissue piece is hollowed out by using a hollow needle with respect to the tissue piece of the sliced tissue piece with a film (iv), and (iv) the tissue piece collected in the hollow needle by repeating this hollowing process And (v) by slicing a slice from a tissue array block in which laminated and fused tissue pieces prepared by heating and fusing the laminate are arranged. After a part of the tissue piece was hollowed out with a hollow needle with respect to the tissue piece of the tissue piece with a film, the hollow needle was cut out when the hollow needle was separated from the film surface with the tissue piece held in the hollow needle. Tissue pieces peel easily from the film surface.

  An example of the method for producing a tissue array sheet of the present invention will be described more specifically below, but the method and apparatus for carrying out the present invention are not limited to this.

  First, as shown in FIG. 1, a film 22 is attached to the surface of the tissue block 21.

  Next, the surface side of the tissue block 21 to which the film 22 is attached is thinly sliced to obtain a tissue piece 23 to which the film 22 is attached. By slicing the tissue block 21 in this manner, the shape of the sliced tissue piece 23 can be maintained by the film 22. Specifically, in this slicing step, the tissue block 21 to which the film 22 is attached is sliced with a sliding mitochrome or rotary mitochrome blade 10.

  Next, as shown in FIG. 2, a hollow needle 18 is inserted into the tissue piece of the tissue piece with film 23 to cut out only the tissue piece 23, and the small piece 25 of the tissue piece 23 is held inside the hollow needle 18. .

  On the lower surface side of the tissue piece 23 with the film 22, this tissue hollowing step may be performed in a state where an elastic material such as a rubber sheet is disposed. Paper may be used instead of an elastic material such as a rubber sheet.

  In this tissue hollowing step, a necessary portion is cut out from the portion of interest of the tissue piece 23 using the hollow needle 18.

  This operation is repeated a plurality of times (the lowering and raising of the hollow needle 18 are repeated), and the plurality of small pieces 25 are stacked inside the hollow needle 18. After the hollow needle 18 is lowered and the tissue piece 23 is cut out, when the hollow needle 18 is raised relative to the tissue piece 23, the cut out tissue piece is easily peeled off from the surface of the film 22.

  In the laminating step, the tissue collected in the hollow needle 18 is laminated by repeating the tissue punching step. The number of repetitions is not particularly limited, but when a tissue piece 23 having a thickness of 100 μm is used, it is preferably 5 to 50 times. The particularly preferred number is 10 to 30 times. Further, the size of the hollow needle 18 is appropriately selected. The hollow needle 18 preferably has an inner diameter of 0.5 to 3 mm and a length of 1 cm to 5 cm. The hollow needle 18 is preferably made of metal.

  Next, the laminated body 27 in the hollow needle 18 is heated and fused. This fusion process can be performed by heating the hollow needle 18 from the outside. Alternatively, as shown in FIG. 3, the laminate 27 can be heated by a heating device 26 provided at one end of the table 5. In this case, the heating device 26 typically includes a heating base 28 having a hole portion 30 and excellent heat conductivity, a heater for heating the heating base 28, a temperature sensor, And a temperature control device having a feedback function. The number of the holes 30 in the heating device 26 may be one or plural. A known device can be used as the heating device 26, and for example, a heater that is electrically heated by a filament can be used. A heat insulating layer is usually provided around the heater.

  A hollow needle including the laminated body 27 can be inserted and arranged in the hole 30 of the heating base 28 and heated. Or as shown in FIG. 4, you may insert and arrange | position the laminated body 27 itself obtained above in the hole 30 of the base 28 for a heating, and may heat it. When heating, the laminated body 27 may be compressed. The heating temperature and time are not particularly limited. The hole 30 may be surrounded by a flowing substrate such as a gel.

  The hollow needle 18 can be heated with warm air without using the heating base 28. When warm air is applied to the hollow needle 18, a warm air blower may be disposed in the vicinity of the hollow needle 18 and hot air of 50 ° C. to 100 ° C. may be applied to the hollow needle 18 for about 1 minute to 2 hours.

  Next, the fusion product obtained by heating the laminated body 27 is cooled in the hollow needle 18 or the heating base 28. When cooling, a cooling device or cold air may be used, or natural cooling may be used. Then, it progresses to the next insertion process.

  In the insertion step, the fusion product 31 is inserted in the holes 33 arranged in the base block 32 in the axial direction (FIG. 5). The base block 32 is a block in which a plurality of cylindrical holes are arranged in an array. The holes can be formed by punching or the like in a separate process. The base block 32 only needs to be made of a material whose surface can be sliced into a sheet shape, and is a paraffin block, for example.

  When the fusion product 31 in the hollow needle 18 is taken out, the fusion product 31 may be pushed out of the hollow needle 18 with a metal rod or wire having a diameter smaller than the inner diameter of the hollow needle 18.

  By repeating the above steps, the holes 33 of the base block 32 are filled with the fusion product 31, and the tissue array block 21 is obtained.

  It is preferable that the fusion 31 is fixed to the base block 32 after the fusion 31 is inserted into the base block 32. Specifically, for example, in a state where the fusion product 31 is inserted into the holes 33 of the base block 32, the base block 32 may be heated to melt the paraffin contained in the film. Alternatively, the base block 32 is configured so that a space remains in the pores of the base block 32 even after the fusion product 31 is inserted. After the fusion product 31 is inserted, the space is filled with dissolved paraffin, and then the paraffin is added. May be solidified.

  As shown in FIGS. 6 (a) to 6 (b), the fusion product 31 is inserted into the air holes 33 of the base block 32, fixed as necessary, and then crossed over the fusion product 31. Slice 32. When slicing the base block 32, the base block 32 is preferably cooled to, for example, −10 ° C. or lower by a cooling device.

  A well-known thing can be used for a cooling device. For example, a method using a refrigerant such as liquid nitrogen, a cooling method using dry ice, or the like can be used.

  You may provide this cooling device in the edge part of the table 5 of the manufacturing apparatus of the tissue array sheet | seat mentioned later.

  The tissue array sheet 11 is obtained by slicing the tissue array block produced by the above process into several μm (usually 4 μm to 6 μm) (FIG. 6B).

  By mounting the tissue array sheet 11 on a substrate 12 such as a slide glass or a nylon film, a tissue array chip in which laminated / fused tissue pieces are arranged can be produced (FIG. 6C). Note that the size of the tissue array chip is arbitrary, but if each tissue array chip just enters the business card holder, the management and organization of the tissue array chip becomes easy.

  Next, an example of the tissue array sheet manufacturing apparatus will be described.

  As shown in FIG. 7 to FIG. 10, the manufacturing apparatus A includes a base 2 formed in a rectangular shape and horizontal X-axis and Y-axis directions placed on the base 2 and operated by moving means 4. A table 5 movable to the base 2, an elevating means 6 attached to the base 2 so as to be movable up and down, and a tissue mounted on the table 5 by the lowering of the elevating means 6 attached to the elevating means 6 Tissue hollowing means 8 is provided to pierce a piece with a hollow needle 18 and cut out a part of the piece of tissue.

  The moving means 4 includes a support base 42 fixed to the base 2, a first moving body 46 screwed into a first rotating shaft 44 passed through the support base 42, and the first moving body. A second rotating shaft 45 passed through 46, and a second moving body 48 disposed on the first moving body 46 and screwed into the second rotating shaft 45. The table 5 is slidably disposed on the second moving body 48. Therefore, the table 5 can move in the XY directions with respect to the support base 42.

  That is, two concave grooves (not shown) are formed on the surface of the second moving body 48 along the longitudinal direction thereof, and a protrusion (not shown) formed on the lower surface of the table 5 is formed into the concave grooves. Is slidably fitted. The table 5 is formed of a rectangular parallelepiped member, and the surface of the table 5 is formed in a rectangular shape. In this embodiment, the table 5 has a central position (FIG. 7), a right position, and a left position (FIG. 8) with respect to the second moving body 48 in the front view shown in FIG. It is configured to be lockable at three locations.

  On the table 5, an area (material stand 52) in which a tissue piece to which a film is attached is arranged at the center position. As described above, the tissue piece to which the film is attached is obtained by thinly slicing the surface side of the tissue block 21 to which the film is attached. The material table 52 may include a sandwiching unit for sandwiching the tissue piece to which the film is attached. When the document table 52 includes the clamping portion, when the tissue piece 23 is cut out with the hollow needle 18, the movement of the tissue piece 23 can be suppressed, and the target portion of the tissue can be cut out. When the hollow needle 18 is lifted after being hollowed out by the needle 18, it is possible to prevent the tissue piece with a film from moving upward as the hollow needle 18 moves.

  A heating device 26 may be provided at one end of the table 5. A cooling device 29 may be disposed at the other end of the table 5. A blade for thinly slicing the surface of the tissue array block may be disposed in the vicinity of the cooling device 29.

  The elevating means 6 includes two support columns 60, 60 erected from the base 2, a holding frame 62 that holds the support column 60, an elevating table 64 that is externally fitted to the support column 60, and an elevating table 64. And a spring 68 that is disposed between the base 2 and urges the lifting platform 64 upward, and a handle 66 that is rotatably attached to the upper end of the holding frame 62. By rotating this handle 66, the upper end of the lifting platform 64 can be pressed and the lifting platform 64 can be lowered by a predetermined dimension.

  The tissue hollowing means 8 includes a hollow needle 18 whose tip is pierced into the tissue piece 23, a cylindrical body 16 arranged in alignment with the hollow needle 18, and the hollow needle 18 and the cylindrical body 16. And an extrusion pin 20 to be inserted into the. The hollow needle 18 may or may not be configured to be turned upside down. The cylindrical body 16 may or may not be configured to be swingable in the horizontal direction.

  As shown in FIG. 9 and FIG. 10, a rotating table 70 is rotatably attached to the front surface of the lifting table 64 around an axial center in the front-rear direction, and a hollow needle 18 is attached to the tip of the rotating table 70. The hollow needle 18 can be easily replaced with a plurality of types (those with different diameters and / or lengths) and attached to the tip of the turntable 70. The turntable 70 is configured to be able to be fixed to the lifting table 64 and to be unlockable. In the figure, reference numeral 72 denotes a fixing release mechanism for the turntable 70.

  Further, on the upper side of the turntable 70, a swing table 74 is attached to the lift table 64 so as to be swingable around a vertical axis, and the cylindrical body 16 is attached to the tip of the swing table 74. ing. In the figure, reference numeral 75 denotes a swing release mechanism for the swing base 74.

  When the hollow needle 18 needs to be turned upside down, after the rocking table 74 is swung left and right to a position that does not interfere with the rotation of the rotating table 70 (FIG. 7), the rotating table 70 is rotated 180 degrees. Thus, the hollow needle 18 can be reversed. Inversion of the hollow needle 18 is not essential for the implementation of the present invention. The hollow needle 18 and the cylindrical body 16 are formed with substantially the same diameter. In a state in which the axial centers of the hollow needle 18 and the cylindrical body 16 are aligned, a gap is formed between the upper end portion of the hollow needle 18 and the lower end portion of the cylindrical body 16. By operating the handle 66 and lowering the lifting platform 64 in a state where the hollow needle 18 and the cylindrical body 16 are aligned with each other, the hollow needle 18 and the cylindrical body 16 are both moved downward to be hollow. The lower end of the needle 18 enters the tissue piece 23 placed on the table 5.

(Positioning means for heating laminated and fused tissue pieces)
The apparatus may or may not include positioning means for heating the laminated / fused tissue piece. When the positioning means 14 is provided, the positioning means 14 for inserting the laminated body (or the hollow needle including the laminated body) into the hole 30 of the heating base 28 includes a plurality of positioning recesses attached on the base 2. A tip 78 having 76, an engaging portion 80 provided on the second moving body 48 of the moving means 4 and capable of engaging with the recess 76 formed in the tip 78, the moving means 4 and the table 5 The means for fixing the heating base 28 placed on the chip to a corresponding position and the engaging portion 80 engaged with the concave portion 76 on the chip 78 are moved to the adjacent concave portion 76 on the chip 78. And adjusting means 40 that can move the moving means 4 as described above.

  That is, the chip 78 is detachably attached to the chip holding frame 62 fixed to the front end of the support base 42. The chip 78 is formed of a steel plate or the like, and a plurality of recesses 76 are formed at a predetermined interval on at least the upper surface of the chip 78. Note that a guide groove for connecting the adjacent recesses 76 may be formed on the upper surface of the chip 78.

  The engaging portion 80 is provided at the tip of an engaging member 82 that protrudes from the front end surface of the second moving body 48. The engaging portion 80 includes a ball that can be engaged with the recess 76 of the chip 78 and a spring that elastically presses the ball. By operating the adjusting means 40 to engage the engaging portions 80 into the respective concave portions 76, the positions of the hole portions 30 formed in the heating base 28 are moved corresponding to the respective concave portions 76. It is configured.

  The moving means 4 is provided with an adjusting means 40 that can move the engaging means 80 so as to move the engaging portion 80 engaged with the concave portion 76 on the chip 78 to the concave portion 76 on the adjacent chip 78. A rotating shaft and a dial that rotates the rotating shaft are included. The rotating shaft may be the first rotating shaft 44 and the second rotating shaft 45 described above. The rotating shaft is provided with a thread groove. The dial 40 is fixed to the ends of the rotary shafts 44 and 45. That is, by operating the dial 40, the table 5 moves in the horizontal direction of the X axis and the Y axis.

(Positioning means for lamination / fusion tissue piece insertion into the base block)
The apparatus may or may not include positioning means for insertion of the laminated / fused tissue piece into the base block. When the laminated / fused tissue piece is inserted in the hole provided in the base block in the axial direction using the positioning means, for example, the following configuration is used.

  The positioning means 14 can have substantially the same configuration as the positioning means 14 that inserts the laminated body (or the hollow needle including the laminated body) into the hole 30 of the heating base 28.

  That is, the positioning means 14 includes a chip 78 having a plurality of positioning recesses 76 mounted on the base 2 and a recess 76 formed in the chip 78 provided on the second moving body 48 of the moving means 4. An engaging portion 80 that can be engaged, a means for fixing the moving means 4 and the base block placed on the table 5 in corresponding positions, and an engagement engaged with the recess 76 on the chip 78. Adjustment means 40 capable of moving the moving means 4 so as to move the joint portion 80 to the adjacent concave portion 76 on the chip 78.

  The chip 78 is detachably attached to a chip holding frame 62 fixed to the front end of the support base 42. The chip 78 is formed of a steel plate or the like, and a plurality of recesses 76 are formed at a predetermined interval on at least the upper surface of the chip 78. Note that a guide groove for connecting the adjacent recesses 76 may be formed on the upper surface of the chip 78.

  The engaging portion 80 is provided at the tip of an engaging member 82 that protrudes from the front end surface of the second moving body 48. The engaging portion 80 includes a ball that can be engaged with the recess 76 of the chip 78 and a spring that elastically presses the ball. By operating the adjusting means 40 to engage the engaging portions 80 into the respective recesses 76, the positions of the holes provided in the base block corresponding to the respective recesses 76 are moved. .

  The moving means 4 is provided with an adjusting means 40 that can move the engaging means 80 so as to move the engaging portion 80 engaged with the concave portion 76 on the chip 78 to the concave portion 76 on the adjacent chip 78. A rotating shaft and a dial that rotates the rotating shaft are included. The rotating shaft may be the first rotating shaft 44 and the second rotating shaft 45 described above. The rotating shaft is provided with a thread groove. The dial 40 is fixed to the ends of the rotary shafts 44 and 45. That is, by operating the dial 40, the table 5 moves in the horizontal direction of the X axis and the Y axis.

  Next, a method of using the tissue array sheet manufacturing apparatus of the present invention will be described.

  The tissue piece with film 23 is arranged on the data base 52 at the center of the table 5, and the periphery of the tissue piece with film 23 is clamped by the clamping unit. A chip 78 having a recess 76 having an arrangement corresponding to the arrangement of the hole 30 formed in the heating base 28 may or may not be attached to the chip holding frame 62.

  Next, the handle 66 of the lifting / lowering means 6 is lowered, and the hollow needle 18 of the tissue punching means 8 is inserted into the tissue piece 23 as shown in FIG. The small piece 25 is accommodated and held. By repeating this operation a plurality of times, a plurality of small pieces 25 of tissue pieces are stacked in the hollow needle. As a result, the laminated body 27 is held in the hollow needle 18.

  Next, the table 5 is moved from the position shown in FIG. 12A to the position shown in FIG. 12B, and the heating base 28 arranged on the table 5 is arranged below the hollow needle 18. And (1) the hollow needle 18 itself including the laminated body 27 is disposed in the hole 30 of the heating base 28, or (2) the push pin 20 is inserted into the hollow needle 18 and the cylindrical body 16. Then, the laminated body 27 is extruded, and the laminated body 27 is disposed in the hole 30 of the heating base 28.

  By heating the heating base 28, the laminate 27 is fused, and a fusion product 31 in which a large number of tissue pieces are fused with each other is obtained.

  This fusion 31 is inserted into the holes 33 of the base block 32 to obtain a tissue array block. The tissue array sheet manufacturing apparatus may include a cooling device 29. When the cooling device 29 is provided, the tissue array block can be provided at the other end of the table.

  As shown in FIGS. 6B to 6C, after cooling the tissue array block 21 by the cooling device 29, the surface of the tissue array block 21 is thinly sliced. By mounting the obtained tissue array sheet 11 on a substrate 12 such as a slide glass or a nylon film, a tissue array chip in which laminated / fused tissue pieces are arranged is obtained.

  Here, since the position of the concave portion 76 of the chip 78 and the position of the hole 30 of the heating base 28 correspond to each other, the adjusting portion 40 moves the engaging portion 80 engaged with each concave portion 76 of the chip 78. By doing so, the hole 30 corresponding to the concave portion 76 of the tip 78 is positioned immediately below the hollow needle 18. Therefore, by repeating the above operation, the tissue can be reliably and easily disposed in the hole 30 of the heating base 28 and in the hole 33 of the base block 32.

  When the laminated body is fused in the hollow needle 18 and / or the cylindrical body 16 and this fusion product 31 is inserted into the hole 33 of the base block 32 arranged on the cooling device, the hollow needle 18 and the cylindrical body are inserted. After inserting the extruding pin 20 into the body 16 from the upper side to extrude the fusion 31 and aligning it by the positioning means 14, the fusion 31 is buried in the holes 33 of the base block 32, and the tissue An array block 21 is obtained.

(Another embodiment)
In the above embodiment, the fusion product 31 is inserted into the hole 33 of the base block 32 to produce a tissue array block. However, as shown in FIG. The fusion 31 may be disposed in the hole 56. In this case, the fusion 31 is placed in the embedding dish 54 through the through-hole 56 of the pallet 55, and the molten paraffin liquid is poured into the embedding dish 54, so that the fusion 31 and the paraffin are integrated. A tissue array block is obtained. The obtained tissue array block is sliced with a microtome to obtain a section (tissue array sheet).

(Tissue piece storage)
By using the film 22 of the present invention, a tissue piece storage device 43 can be produced.

The tissue piece storage device includes, for example, as shown in FIG. 15, (a) a film 22, (b) a tissue piece 47, and (c) a base material 47, where (b) the tissue piece 23 is (A) It is located between the film 22 and the (c) base material 47, and (b) the surface of the film 22 in contact with the tissue piece 23 has adhesiveness to the tissue piece 23. This adhesiveness is achieved by applying an acrylic pressure-sensitive adhesive to the surface of the film 22 (a) in contact with the tissue piece 23 (b). In addition, the (a) film 22 is preferably a film 22 that can be used for manufacturing the array sheet of the present invention, that is,
(A) Adhesiveness to the tissue piece 23 that adheres to the surface of the tissue piece 23 and maintains the attached state;
(B) In the manufacture of the array sheet, after part of the tissue piece 23 is hollowed out by a hollow needle, the moldable tissue piece 23 can be released;
(C) the ability to retain the shape during the operation of manufacturing the array sheet; and
(D) In the manufacture of the array sheet, strength that is not cut when a part of the tissue piece 23 is cut out by a hollow needle;
Is provided.

  The storage device 43 can be manufactured by using the tissue piece 23 to which the film 22 obtained above is attached and attaching a base material 47 to the tissue piece 22.

  The tissue piece 23 affixed to the film 22 obtained by peeling the base material 47 from the storage device 43 can be used for manufacturing the array sheet of the present invention.

  Further, when the storage device is not directly used for manufacturing the array sheet, the features (b) to (d) are not necessarily provided.

  The pressure-sensitive adhesiveness (a) and the releasability (b) are achieved, for example, by applying an adhesive to the surface of the film 22 to be attached to the tissue piece 23. Examples of such an adhesive include, but are not limited to, acrylic pressure-sensitive adhesives.

  The shape retention ability of (c) and the strength of (d) are achieved, for example, by producing the film 22 with a material selected from the group consisting of polyethylene terephthalate, polyvinyl chloride, paper, and polypropylene film. When using paper, the surface of the paper may or may not be mirror-finished.

  The strength of the above (d) can be easily achieved by adjusting the thickness of the film 22. The thickness of the film 22 can be appropriately determined by those skilled in the art. Typically, in the case of a film selected from the group consisting of polyethylene terephthalate, polyvinyl chloride, paper, and polypropylene film, 20 μm to 200 μm. It is about the thickness.

  The main component of the above-mentioned (a) base material 47 is not limited. For example, the main component can be a substance selected from the group consisting of polyethylene terephthalate, polyvinyl chloride, paper, and polypropylene film. .

  As a result, the storage device of the present invention can be used as a tissue bank. In the storage device 43 of the present invention, the film 22 preferably covers the entire tissue piece 23 and blocks the tissue piece 23 from the outside of the storage device. In this case, since the tissue piece 23 contained in the storage device of the present invention is not exposed to the air outside the storage device, the nucleic acid and protein contained in the tissue piece 23 in the storage device are denatured or decomposed. The tissue piece 23 can be stored for a long time without (or only undergoing a small degree of denaturation or degradation).

  In the storage device 43 of the present invention, information on the tissue piece 23 included in the storage device 43 may be described. For example, as shown in FIG. 16, the storage device 43 of the present invention may be placed in a folder 49 such as a business card folder, and the folder 49 may be stored in a cool dark place or in a frozen state. In this case, the information on the tissue piece 23 included in the storage device 43 may be described directly in the storage device 43 or may be described in the folder 49 that stores the storage device 43. Thus, a tissue bank can be created by maintaining the correspondence between the information on the tissue piece 23 included in the storage tool 43 and the storage tool 43.

  And the base material 47 is peeled from the preservation | save tool 43 as needed, and the tissue piece 23 affixed on the film 22 can be utilized for manufacture of the array sheet of this invention.

  It can be used for histological diagnosis, and special staining and immunostaining can also be determined.

A tissue array sheet manufacturing apparatus 2 base 4 moving means 5 table 6 elevating means 8 hollowing means 10 heating base 12 substrate 14 positioning means 16 cylindrical body 18 hollow needle 20 push pin 21 tissue block 22 film 23 tissue piece 25 Small tissue piece 26 Heating device 27 Laminate 28 Heating base 29 Cooling device 30 Heating base hole 31 Fusion 32 Base block 33 Base block hole 43 Preserving tool 47 Base 49 Folder

Claims (17)

(A) attaching a film to the surface of the tissue block;
(B) slicing the surface side of the tissue block to which the film is attached to obtain a tissue piece to which the film is attached;
(C) piercing a hollow needle into the tissue piece of the tissue piece with film, and holding the small piece of the tissue piece inside the hollow needle;
(D) repeating the above step (c) to obtain a laminate comprising a plurality of small pieces inside the hollow needle;
(E) heating the laminate to obtain a fusion;
(F) inserting the fusion into a hole provided in the base block;
(G) slicing the base block across the laminate;
For producing a tissue array sheet comprising: (A) attaching a film to the surface of the tissue block;
(B) slicing the surface side of the tissue block to which the film is attached to obtain a tissue piece to which the film is attached;
(C) piercing a hollow needle into the tissue piece of the tissue piece with a film, and holding a small piece of the tissue piece inside the hollow needle;
(D) repeating the above step (c) to obtain a laminate comprising a plurality of small pieces inside the hollow needle;
(E) heating the laminate to obtain a fusion;
(F) The fusion product is placed in a through-hole of a pallet arranged in the embedding dish, and a molten paraffin solution is poured into the embedding dish to obtain a tissue array block in which the fusion product and paraffin are integrated. Process,
(G) slicing the tissue array block across the fusion;
For producing a tissue array sheet comprising: (A) attaching a film to the surface of the tissue block;
(B) slicing the surface side of the tissue block to which the film is attached to obtain a tissue piece to which the film is attached;
(C) piercing a hollow needle into the tissue piece of the tissue piece with film, and holding the small piece of the tissue piece inside the hollow needle;
(D) repeating the above step (c) to obtain a laminate comprising a plurality of small pieces inside the hollow needle;
(E) heating the laminate to obtain a fusion;
(F) inserting the fusion into a hole provided in the base block;
For producing a tissue array block comprising: (A) attaching a film to the surface of the tissue block;
(B) slicing the surface side of the tissue block to which the film is attached to obtain a tissue piece to which the film is attached;
(C) piercing a hollow needle into the tissue piece of the tissue piece with a film, and holding a small piece of the tissue piece inside the hollow needle;
(D) repeating the above step (c) to obtain a laminate comprising a plurality of small pieces inside the hollow needle;
(E) heating the laminate to obtain a fusion;
(F) placing the fusion in a through-hole of a pallet arranged in the embedding dish, pouring molten paraffin liquid into the embedding dish, and integrating the fusion and paraffin;
For producing a tissue array block comprising:   The method for producing a tissue array block according to any one of claims 1 to 4, wherein the film is one selected from the group consisting of polyethylene terephthalate, polyvinyl chloride, paper, and polypropylene film. A base, and a table placed on the base and movable in the horizontal X-axis and Y-axis directions by operating the moving means;
Elevating means attached to the base and arranged to be elevable;
A tissue punching means for punching out the tissue piece of the tissue piece with a film attached to the lifting means and placed on the table by the lowering of the lifting means;
Means for punching the tissue piece of the tissue piece with a film a plurality of times by the tissue cutting means to obtain a laminate;
A heating device for heating the laminate;
An apparatus for manufacturing a tissue array sheet.   The apparatus according to claim 6, wherein the heating means is disposed on a table.   The heating means includes a heating base having a hole and excellent heat conductivity, a heater for heating the heating base, a temperature sensor, and a temperature control device having a feedback function. The device of claim 6.   The apparatus according to claim 6, wherein the heating means is a hot air blower that gives warm air to the hollow needle.   The apparatus according to claim 6, further comprising cooling means disposed on the table.   A film used for producing a tissue array sheet, the film sticking to the surface of the tissue block, and also sticking to the tissue block after slicing the tissue piece with film from the tissue block A film having adhesiveness to the tissue block.   It is a film of Claim 11, Comprising: The film by which the acrylic adhesive is apply | coated to the surface stuck on the surface of the said tissue block.   12. A film according to claim 11, comprising as a main component a substance selected from the group consisting of polyethylene terephthalate, polyvinyl chloride, paper, and polypropylene film. Tissue piece storage device, including:
(A) film;
(B) a tissue piece; and (c) a substrate,
Here, the (b) tissue piece is located between the (a) film and the (c) substrate,
The (b) preservation | save implement in which the surface of this (a) film which contact | connects a tissue piece has adhesiveness with respect to a tissue piece.   The storage device according to claim 14, wherein an acrylic adhesive is applied to a surface of the (a) film in contact with the (b) tissue piece.   15. The storage device according to claim 14, wherein the (a) film is mainly composed of a substance selected from the group consisting of polyethylene terephthalate, polyvinyl chloride, paper, and polypropylene film.   The storage device according to claim 14, wherein the (a) base material is mainly composed of a substance selected from the group consisting of polyethylene terephthalate, polyvinyl chloride, paper, and polypropylene film.