JPH0943118A - Tissue treating device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To treat a tissue sample adhering to a slide glass with a reagent by controlling the using amount of the reagent as small as possible. SOLUTION: A cover assembly 60 is held in the well 56 of a reagent treating unit 54 in a vertically movable state. The assembly 60 is provided with a reagent receiver 62 having a reagent groove 61 and a cover plate 12 energized in a inclining direction through a bent section 64. The cover plate 12 is provided with a meshed sheet 18 and, when a slide glass 36 is lowered, the meshed sheet 18 is wetted with a reagent sucked in the very small clearance between the cover plate 12 and the slide glass 36 and the reagent is supplied to a sample on the plate glass 36, because the plate 12 is brought into contact with the glass 36 due to a slope 66, namely, the sample is covered with the meshed sheet 18.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a biological tissue processing apparatus, and more particularly to a structure for supplying a minimum amount of reagents to a sample of biological tissue.

[0002]

2. Description of the Related Art Along with recent advances in biotechnology, various treatments for biological tissues are being actively carried out at research facilities, universities, etc., and there is a strong demand for efficient and speedy treatment. Has been done. For example, immunostaining or ISH (in
In the case of performing in situ hybridization) processing, a thin slice of biological tissue (hereinafter simply referred to as a sample) is attached on a slide glass serving as a transparent sample plate, and the sample on the slide glass is dehydrated, stained, Many processes such as hybridization are performed.

In the case of performing such a treatment, conventionally, a large number (for example, 20 sheets) of slide glasses to which a sample is attached are generally arranged in a metal cage at regular intervals and kept standing upright. The basket was dipped in a large glass reagent tank, and such treatment was performed on various reagents.

[0004]

However, in the above-mentioned conventional processing method, there is a problem that it is complicated because the injection of the reagent into the reagent tank and the removal of the reagent after the processing are artificially performed. However, since it is necessary to use a large reagent tank that can immerse a large number of glass slides that are spaced apart from each other, the amount of each reagent must be uniformly large. is there. That is, it is necessary to use a large amount of reagents in the processing such as deparaffinization, but on the other hand, there are many expensive reagents that are sufficiently small in quantity, and the cost cannot be reduced as a whole processing, resulting in poor economic efficiency. Then, for the same reason, there is a problem that the amount of drained liquid becomes larger than necessary.

Further, conventionally, since the same reagent treatment is performed collectively for each sample on the slide glass in the basket, it is necessary to perform different reagent treatment for some samples from the middle of the treatment process. In that case, it was necessary to remove a part of the slide glass from the basket and perform the above treatment separately.

Incidentally, in the past, in order to reduce the amount of reagent and realize individual reagent treatment, a sample of a slide glass was covered with a cover glass, and a reagent was dropped between the slide glass and the cover glass. It has also been practiced to use the surface tension of the reagent to take in the reagent between the glasses and to carry out the reagent treatment, but it is easy for bubbles to form between the glasses due to stains, etc. However, there is also a problem that work efficiency is poor. Also,
Conventionally, it has also been proposed to perform reagent treatment on a sample by placing a slide glass and another plate in close proximity to each other to form a minute gap between them and supplying a reagent from above or below the minute gap. However, it is difficult to accurately form such a uniform minute gap, and there is a problem that the structure becomes complicated.

The present invention has been made in view of the above conventional problems, and an object of the present invention is to provide a biological tissue processing apparatus capable of performing processing using a minimum required amount of reagents and preventing waste of the reagents. To provide.

Another object of the present invention is to efficiently supply the reagent over the entire area of the sample on the sample plate (slide glass).

[0009]

In order to achieve the above object, the present invention provides an apparatus for performing a reagent treatment on a sample of biological tissue attached on a sample plate, which is bonded to the sample attachment surface of the sample plate. It is characterized by including a cover plate having a mesh sheet provided on its surface and a reagent supply means for supplying a reagent to the mesh sheet.

According to the above structure, the cover plate having the mesh sheet is joined to the sample adhering surface of the sample plate. As a result, the mesh sheet is interposed between the sample plate and the cover plate, and a minute gap is formed between the two plates containing the sample. At the same time, the entire sample is covered with the mesh sheet. Desirably, the mesh sheet is formed of a mesh or is composed of a thin and flexible sheet having porosity. The presence of such a thin mesh sheet between the two plates can cause the capillary action of the reagent between the two plates, that is, the surface sheet allows the reagent to permeate through the mesh sheet as a medium. You can Such surface tensions occur in the microgaps between the two plates and in the microvoids in the mesh sheet itself. In any case, the sample in contact with the mesh sheet will be in a reagent wet state, and the reagent treatment of the sample will be achieved. Since the minute gap can be formed between the two plates only by joining the two plates with the mesh sheet interposed, delicate positioning and high-precision processing are unnecessary.

[0011] In a preferred aspect of the present invention, the sample plate and the cover plate include holding means for holding both in a joined state, and the reagent supply means includes the plate joining held by the holding means. It consists of a reagent bath into which the lower end of the body is immersed.

That is, when the lower end of the plate assembly is immersed in the reagent tank containing the reagent, the reagent is taken in by the surface tension from the minute gap exposed at the lower end or the end of the mesh sheet, and the reagent is introduced into the sample. Can be supplied.

Further, in order to achieve the above object, the present invention is an apparatus for treating a sample of biological tissue attached on a sample plate with a reagent, the cover plate having a mesh sheet, and the cover plate. A cover assembly configured to swingably support a lower end portion of the cover plate, and a reagent tank to which a lower end portion of the cover plate is guided to supply a reagent to the mesh sheet, and a cover assembly including the cover assembly. When a sample plate is set, a rocking drive unit that raises the cover plate in an inclined state and joins the mesh sheet of the cover plate to the sample attachment surface of the sample plate, To do.

According to the above construction, the cover assembly is mainly composed of the cover plate and the reagent tank to which the lower end portion is guided, and the swing support portion is formed at the lower end portion of the cover plate, and the cover plate is concerned. Is swingably supported. Since the lower end of the plate joined body is in contact with the reagent in the reagent tank, the reagent is sucked between the plates due to the surface tension and spreads over the entire mesh sheet, and the mesh sheet is kept in a reagent wet state. In this case, it is desirable to extend the mesh sheet to the lower end of the cover plate so that the lower end of the mesh sheet directly contacts the reagent. However, if the reagent is smoothly sucked into the inside due to the capillary phenomenon of the minute gap between the plates, the mesh sheet can be put into the reagent wet state without necessarily extending and exposing the mesh sheet to the lower end. .

When the sample plate is set on the cover assembly, the swing drive unit raises the cover plate from the inclined state, whereby the mesh sheet of the cover plate is joined to the sample attachment surface of the sample plate. That is, the cover plate is configured to be openable and closable,
When the sample plate is set, the cover plate is closed and the two plates are joined. As a result of this joining, a minute gap is formed between the two plates as described above, the sample is taken from the reagent tank into the minute gap, and at the same time, the reagent is supplied to the mesh sheet, and the sample covered with the mesh sheet is Reagent processing is performed.

[0016] In a preferred aspect of the present invention, the swing drive unit receives an assembly elevating mechanism for vertically moving the cover assembly on which the sample plate is set, and a lowering direction of the cover assembly. And a well for erecting the cover plate in an inclined state by utilizing the well.

Further, in a preferred aspect of the present invention, the well has a sloped surface which widens an opening upward,
The cover plate is erected by the inclined surface as the cover assembly descends. That is, when the elevating mechanism lowers the sample plate from above and inserts it into the cover assembly, the entire cover assembly descends in the well, at which time the cover plate rises vertically due to the slope. As a result, the cover plate is joined to the sample plate. That is, a part of the downward movement of the sample plate can be converted into the closing movement of the cover plate.

In a preferred aspect of the present invention, a lower end of the cover plate is provided with a biasing means for imparting an inclined biasing force to the cover plate, and preferably the biasing means is composed of a leaf spring. It

[0019]

Preferred embodiments of the present invention will be described below with reference to the drawings.

FIG. 1 illustrates the principle of the present invention.
In FIG. 1, a cover plate 12 is joined to a sample attachment surface side of a slide glass 10 as a transparent sample plate to which a sample of biological tissue is attached. The holder 14 holds the upper end of the joined body of the two plates.

The cover plate 12 is composed of a base plate 16 as a substrate and a mesh sheet 18 provided on one surface of the base plate 16. The base plate is made of a Teflon plate material having a thickness of 1 mm, for example. The vertical and horizontal dimensions are the same as those of the slide glass 10. The mesh sheet 18 is made extremely thin to form a capillary gap, and its thickness is, for example, 10
It is set to 0 μm or less and has the same vertical and horizontal dimensions as the slide glass 10 like the base plate 16. That is, the mesh sheet 18 is interposed between the slide glass 10 and the base plate 16 to form a so-called sandwich structure.

The mesh sheet is a thin mesh-like sheet having porosity, and is made of nylon, for example, and has a thickness of 40 μm in this embodiment. When nylon is used, the thickness of the nylon fiber is, for example, φ40 μm, and the aperture ratio is, for example, 50%. The mesh sheet 18 is easy to adapt to the reagent,
In addition, it is composed of a material that does not chemically affect the sample. In addition to nylon, a mesh sheet such as Teflon may be used. Since the mesh sheet 18 having such a uniform thickness is interposed between the slide glass 10 and the cover plate 12, a very small gap that causes a capillary phenomenon between the slide glass 10 and the cover plate 12. Are formed, and the reagent is sucked into the gap.

That is, the reagent tank 20 is arranged below the joined body held by the holder 14, and the reagent 22 is placed inside the reagent tank. In such a state, when the joined body supported by the holder 14 is pulled down from the upper side while maintaining the vertical standing state and the lower end portion of the joined body is penetrated into the reagent 22, the slide glass 10 and the cover plate. Reagent 22 in the minute gap between
Are sucked in, that is, the reagent permeates into the mesh sheet 18, and the whole mesh sheet 18 is in a reagent wet state.

FIG. 2 schematically shows such a state. A sample 24 as a thin slice of biological tissue is attached to the slide glass 10, and the entire area of the sample 24 is covered with a mesh sheet 18 in a reagent wet state. In such a state, the reagent 22 becomes the sample 24
Will be sufficiently supplied to the reagent and the reagent treatment will be efficiently performed.

Slide glass 10 and cover plate 12
The suction of the reagent into the minute gap between the slide glass 10 and the cover plate 12 is caused by the action of the surface tension of the minute gap itself between them and the minute gap of the mesh sheet 18 itself. A thin reagent layer is formed between and. With such a mesh sheet 18, a reagent can be supplied to the sample 24 without generating bubbles and the like, and such a reagent treatment can be performed with a necessary minimum amount of reagent.

After the reagent treatment is completed, the joined body is pulled up by the holder 14, the slide glass 10 and the cover plate 12 are removed from the holder 14, and a new cover plate 12 is joined to the slide glass 10 to make the holder. After being set to 14, the same processing as described above will be performed on the next reagent.

It is considered that a thin reagent layer is formed between the sample 24 and the mesh sheet 18 in the reagent wet state. When the above-mentioned materials are used, the sample 24 adheres to the mesh sheet 18 and slides. There is no problem that the sample 24 comes off the glass 10. Further, in the embodiment shown in FIG. 1, the mesh sheet 18 is formed over the entire area of the one surface of the cover plate 12, but of course, the mesh sheet 18 is provided at least in a portion that covers the entire area where the sample exists. If so, the same effect as described above can be obtained. In this case, mesh sheet 1
If the lower end of 8 extends to the lower end of the base plate 16, it is considered that the reagent 22 can be sucked in more smoothly.

Next, a biological tissue processing apparatus to which the principle shown in FIG. 1 is applied will be described with reference to FIG.

FIG. 3 shows a preferred embodiment of the biological tissue treating apparatus according to the present invention, and FIG. 3 is an external view thereof.

A plurality of reagent containers 2 are provided on the upper surface of the apparatus body 26.
8 are arranged upright. Reagents different from each other are stored in the respective reagent containers, and the reagent is dispensed by the dispensing nozzle 30. The dispensing nozzle 30 is configured to be freely movable in three dimensions by the transport mechanism 32, that is, in the X direction, the Y direction, and the Z direction.

On the other hand, the holding mechanism 34 holds a plurality of slide glasses 36 with the longitudinal direction oriented vertically. Specifically, the upper end portion of the slide glass 36 is held by the holding mechanism 34, and a plurality of slide glass rows configured by the plurality of slide glasses 36 are held in an array. The holding mechanism 34 is vertically movable by an upper and lower stage 46.

On the other hand, the processing unit 38 and the processing unit 40, which are horizontally movable by the horizontal stage 48, have a plurality of well-shaped reagent tanks 42 and 44, respectively. In each processing unit 38 and 40, the holding mechanism 3
The plurality of reagent vessels 42 and 44 are formed in the arrangement pattern of the plurality of slide glasses 36 held by 4, and the reagent treatment can be performed on the plurality of slide glasses 36 at one time. However, each reagent tank 42 and 4
4 are separated from each other, and different reagents can be injected by the dispensing nozzle 30 to perform reagent processing.

The reagent processing unit 38 and the reagent processing unit 40 have the same structure as each other, and while the reagent processing is actually performed on the one hand, the preparation for the next reagent processing on the other hand, that is, from the reagent tank is performed. The reagent is removed and the next reagent is dispensed.

Extraction of such a reagent is carried out by a plurality of suction nozzles 50 supported by a vertical stage 49 so as to be vertically movable. The suction nozzles 50 are arranged at the arrangement pitch of the reagent tank, and suction can be performed for each row of the reagent tank.

The reagent tank is formed in the shape of a well having a rectangular cross section, and in the state where the slide glass 36 is inserted in each of the reagent tanks 42 and 44, the upper and lower stages 46 are arranged.
The periodic movement of the slide glass 36 with respect to the slide glass 36 improves the reagent processing efficiency.

The temperature control section 52 joined below the reagent processing units 38 and 40 controls the temperature of each reagent processing unit 38 and 40 at a constant temperature, thereby performing reagent processing under a constant temperature condition. be able to.

On the other hand, on the horizontal stage 48, the reagent processing unit 54 to which the principle shown in FIG. 1 is applied is held so as to be movable in the horizontal direction. This reagent processing unit 54
Has three wells 56 corresponding to the three rows of glass slides held by the holding mechanism 34. Each well 56 has a length slightly longer than the entire slide glass row, and its specific structure will be described later. Below the reagent processing unit 54, a temperature controller 58 having the same function as the temperature controller 52 described above is provided.

FIG. 4 shows a specific structure of the reagent processing unit 54. This reagent processing unit 54 is
The well 56 and the cover assembly 60 provided corresponding to each slide glass 36 are included.
Note that the cover assembly 60 is not shown in FIG. 3 for simplification of the drawing.

The cover assembly 60 includes a reagent receiver 62 having a reagent groove 61 for accommodating a reagent, and a cover plate 12 configured to be openable and closable with respect to the reagent receiver 62.
It consists of and. The lower end of the cover plate 12 is a bent portion 64. Specifically, the bent portion 64 has a leaf spring structure and always biases the cover plate 12 in the direction in which it opens. Of course, due to the action of the slope 66 of the well 56, which will be described later, this cover plate 1
2 can be closed and the standing operation can be performed from the inclined state to the upright state.

The lower end of the mesh sheet 18 extends into the reagent groove 61, that is, the mesh sheet 18
A part of the is always immersed in the reagent. The reagent receiver 62 has an independent reagent groove 61 for each cover assembly 60, but a plurality of reagent receivers 62 may be continuously formed integrally between adjacent cover assemblies 60.

The cover assembly 60 is held in the well 56 so as to be movable up and down by a mechanism for holding the cover assembly 60, and the cover assembly 60 is biased upward with respect to the well 56 by a spring spring (not shown). . Here, the upward movement restricts the upward movement at a constant height.

As shown in FIG. 5, the well 56 has a depth that allows the cover plate 12 to be completely erected when the cover assembly 60 is pulled down and housed in the well 56, as shown in FIG. An inclined surface 66 is formed in the upper portion to gradually raise the cover plate 12 from the inclined state. That is, when the slide glass 36 is pulled down from above by the holding mechanism 34 shown in FIG. 3, the lower end portion of the slide glass 36 pushes down the reagent receiver 62, whereby the entire cover assembly 60 is pushed down. Then, the cover plate 12 is erected while the outer surface of the cover plate 12 slides on the slope 66. This state is shown in FIG. In addition, the convex portion 12 is provided on the upper end of the outer surface of the cover plate.
In the state in which A is formed and the cover assembly 60 is completely housed in the well 56, its convex portion 12A contacts the inner surface of the well 56 and the cover plate 12 is firmly bonded to the slide glass 36. . The opening is slightly opened above the well 56 due to the formation of the slope 66, and a receiving space for the holding portion of the holding mechanism 34 is formed. In the joined state as shown in FIG. 5, as shown in FIG. 1, the reagent in the reagent groove 61 is sucked into the minute gap between the slide glass 36 and the cover plate 12, and the mesh existing in the minute gap is present. The reagent will be spread over the entire sheet 18. Therefore, the reagent is supplied to the sample and the reagent processing is realized.

In this embodiment, for example, 100 μl of the reagent is injected into the reagent groove 61, that is, the reagent treatment can be performed with an extremely minute amount.

Of course, when it is necessary to perform reagent processing using a large amount of reagents, the reagent processing unit 3 shown in FIG.
8 or 40 is used for reagent processing.

When the reagent treatment is completed from the state shown in FIG. 5 and the slide glass 36 is pulled up by the holding mechanism 34, the cover assembly 60 is pulled up by the action of a coil spring (not shown). Goes through the reverse process, that is, the outside of the cover plate slides on the slope 66, and gradually begins to incline,
The state returns to that shown in FIG.

After the processing for a certain reagent is completed, FIG.
The reagent in the reagent groove 61 is sucked using the suction nozzle 50, the other reagent is injected into the reagent groove 61 by the dispensing nozzle 30, and the above-described reagent processing is performed again. Of course,
In order to avoid adverse effects due to the mixing of the reagent contained in the mesh sheet 18 and the next reagent, it is desirable to replace the cover assembly 60 or replace only the mesh sheet 18. In this embodiment,
The mesh sheet 18 is attached to the base plate with an adhesive or the like, and the treatment for each reagent is sequentially performed by replacing the entire cover assembly 60.

Although one cover assembly 60 is shown in FIG. 4, a plurality of such cover assemblies 60 are provided in the well 56 corresponding to each slide glass 36.

Incidentally, when the reagent treatment is performed with a relatively large amount of the reagents, the treatment units 38 and 40 are used. In this case, one reagent treatment unit performs the reagent treatment while the other reagent is treated. Aspiration of the remaining reagent and injection of the next reagent are performed in the processing unit,
The efficiency of the process is improved by alternately performing the reagent process and the preparation for the reagent process. In such a series of reagent treatments, when treatment with a small amount of reagent is desired, the treatment unit 54 is used and the reagent treatment is performed by the operation shown in FIG.

Although the bent portion 64 shown in FIG. 4 has a leaf spring structure in this embodiment,
The same operation as described above may be performed by providing a rotating shaft in the bent portion 64 and further providing a spring that produces a biasing action.

Further, in the configuration shown in FIG. 4, the reagent receiver 6
Although the cover plate 12 is provided only on one side of the sample No. 2, when the reagent treatment via a mesh sheet is performed on two slide glasses whose non-sample adhered surfaces are joined to each other, the reagent receiver 62 is provided. It is formed to have a width corresponding to two such slide glasses, and the cover plate 1 is provided on both sides thereof.
2 may be provided, and two cover plates may be joined from both sides of a pair composed of two slide glasses. In this case, two slopes 66 of the well 56 may be formed on the left and right sides corresponding to the two cover plates.

As described above, according to the biological tissue processing apparatus of the present embodiment, it is possible to deal with both a large amount of reagent processing and a small amount of reagent processing, and it is possible to prevent waste of reagents. Further, when the reagent treatment is performed with a small amount of the reagent, the reagent can be surely supplied over the entire area of the sample by using the cover assembly 60 described above, and the reagent treatment is surely performed without the presence of bubbles and the like. There is an advantage that you can.

Incidentally, the slide glass 1 to the holding mechanism 34
The mounting of 0 may be carried out artificially, but the mounting may be carried out, for example, by holding a plurality of slide glasses 10 upright in a rack and lowering the holding mechanism 34 from above to pick up the slide glasses.

[0053]

As described above, according to the present invention,
Reagent processing can be performed using the minimum required amount of reagent,
There is an advantage that the waste of the reagent can be prevented. Further, there is an advantage that the reagent can be smoothly supplied over the entire area of the reagent on the sample plate, and in that case, generation of bubbles can be prevented. Further, according to the present invention, the cover plate can be surely joined to the sample plate by opening and closing the cover plate, and when the cover plate is detached from the sample plate, the cover glass surface direction to the sample The separation of the sample from the sample plate can be prevented without exerting force.

[Brief description of drawings]

FIG. 1 is a diagram for explaining the principle of the present invention.

FIG. 2 is a schematic cross-sectional view showing the action of a mesh sheet.

FIG. 3 is an external view of a biological tissue processing apparatus according to the present invention.

FIG. 4 is a diagram showing a specific configuration of a cover assembly.

FIG. 5 is a view for explaining the operation of the cover assembly.

[Explanation of symbols]

10, 36 glass slides, 12 cover plates,
18 mesh sheet, 16 base plate, 54
Reagent processing unit, 56 wells, 60 cover assembly, 61 reagent groove, 62 reagent receiver, 64 bent portion.

Claims (8)

[Claims] 1. An apparatus for performing a reagent treatment on a sample of biological tissue attached to a sample plate, comprising: a cover plate having a mesh sheet on a surface of the sample plate that is joined to the sample attachment surface; A biological tissue processing apparatus comprising: a reagent supply unit that supplies a reagent to a sheet. 2. The apparatus according to claim 1, further comprising holding means for holding the sample plate and the cover plate in a state where the sample plate and the cover plate are joined to each other, and the reagent supply means is a plate held by the holding means. A biological tissue processing apparatus comprising a reagent tank in which a lower end portion of a joined body is immersed. 3. A device for performing a reagent treatment on a sample of a biological tissue attached on a sample plate, wherein a cover plate provided with a mesh sheet and a rocker for rockably supporting a lower end portion of the cover plate. A cover assembly composed of a dynamic support portion and a reagent tank to which a lower end portion of the cover plate is guided to supply a reagent to the mesh sheet; and a tilted state when the sample plate is set in the cover assembly. 2. A treatment apparatus for biological tissue, comprising: a rocking drive unit that erects the cover plate in (1) and joins the mesh sheet of the cover plate to a sample attachment surface of the sample plate. 4. The apparatus according to claim 3, wherein the swing driving unit receives an assembly elevating mechanism that moves the cover assembly on which the sample plate is set up and down, and receives the cover assembly from above and lowers the assembly. And a well for erecting the cover plate in an inclined state by using the well, and a treatment device for biological tissue. 5. The apparatus according to claim 4, wherein the well has an inclined surface with an opening widened upward, and the cover plate is erected by the inclined surface as the cover assembly descends. An apparatus for treating biological tissue characterized by: 6. The apparatus for treating biological tissue according to claim 5, wherein the lower end portion of the cover plate is provided with an urging means for applying an inclined urging force to the cover plate. 7. The apparatus for treating biological tissue according to claim 6, wherein the urging means comprises a leaf spring. 8. The apparatus for treating biological tissue according to claim 3, wherein the mesh sheet extends to a desired position inside the reagent tank.