JPH0996592A - Treating system for texture of organism - Google Patents

Abstract

PROBLEM TO BE SOLVED: To realize treatment of the texture of organism requiring no transfer of sample cage nor dispensation with nozzle. SOLUTION: A sample texture of organism is set in a treating case 20. In a state where an inverted U-shaped tube 26 communicating with the treating case 20 is put, at the other end thereof, into a container filled with a reagent, an air pump 28 is driven to introduce the reagent into the treating case 20. Flow-out of reagent from the treating case 20 is blocked by the shape of tube 26 and the reagent can be discharged by operating the air pump 28 upon finishing the treatment.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an apparatus for immersing a sample of biological tissue in a reagent for processing.

[0002]

2. Description of the Related Art Conventionally, a sample of biological tissue is treated with a reagent, for example, ISH (in situ hybridization).
The method has been performed by attaching a sample such as a tissue section of an animal or a plant on a slide glass to prepare a preparation specimen, and supplying various reagents to the preparation specimen in order. In recent years, a hole-mount type ISH that uses a small piece of tissue as a sample as it is without the need to prepare a tissue section has become widespread.

FIG. 5 shows such a conventional method for treating biological tissue. As shown in FIG. 5, the microplate 10 has a plurality of reagent treatment tanks (96, for example).
Wells 12 are formed, and a predetermined reagent 14 is injected into each well 12. On the other hand, sample 18
Are stored in the sample basket 16. Here the sample basket 16
At least the lower surface of which is formed in a mesh shape. And
The sample cage 16 containing the sample 18 is allowed to enter the specific well 12 to immerse the sample 18 in the reagent 14, and after the reagent treatment, the sample cage 16 is transferred again into another well 12 to perform the reagent treatment, Such a series of steps was repeated.

[0004]

In the above-mentioned conventional apparatus, the diameter of each well is as small as 9 mm, for example, and it has been difficult to transfer the sample basket quickly and accurately.
In particular, when artificially doing it, the work was extremely complicated. Incidentally, there are, for example, 16 types of reagents used in ordinary ISH, and the same treatment may be repeated several times with the same reagent.

Further, in the above apparatus, it is necessary to dispense the reagent in each well in advance, and in addition, it is necessary to aspirate and discharge the reagent as necessary. Therefore, a separate dispensing mechanism is provided. There was a problem of cost. When performing such work artificially, the complexity is further increased.

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 which does not require transfer of a reagent cage or dispensing with a nozzle.

It is another object of the present invention to provide a biological tissue processing apparatus which can smoothly aspirate and discharge a reagent with a simple structure.

Another object of the present invention is to provide an automated biological tissue processing apparatus which can efficiently perform a series of reagent processing by applying a new reagent supply system.

[0009]

In order to achieve the above object, the present invention provides a treatment container for containing a sample of biological tissue,
A reagent container containing a reagent, a reagent channel having one end connected to a lower portion of the processing container and the other end guided into the reagent container, and an air pump connected to an upper portion of the processing container. The biological tissue is processed by operating the air pump to introduce the reagent into the processing container through the reagent flow path.

According to the above construction, the air pressure in the processing container is reduced by the operation of the air pump, and the pressure difference resulting from this causes the reagent in the reagent container to flow from the other end of the reagent channel to the reagent channel. And is guided to the lower part of the processing container. After the predetermined reagent is introduced into the processing container, the reagent processing is performed on the sample stored in the processing container by maintaining the state. After the reagent treatment, the reagent is discharged from the treatment container by operating the air pump in the reverse direction.

According to the present invention, it is possible to quickly and easily introduce and discharge a reagent without the need for a conventional sample basket or dispensing mechanism. Here, the processing container according to the present invention has the functions of both the conventional sample cage and the reagent tank, and the device configuration can be simplified in this respect as well. Further, according to the present invention, since the reagent can be statically supplied to the sample in the processing container, for example, the adverse effect on the sample that occurs when the reagent is vigorously dropped from the nozzle above the sample It is possible to prevent the problems such as the scattering of the reagent, the ejection of the sample, and the adhesion of the sample to the inner wall.

[0012] In a preferred aspect of the present invention, the reagent flow path is formed by extending upward from the one end, bending in the shape of an inverted U in the middle, and then extending downward to form the other end. The other end is formed at least at a position lower than the one end. Further, the vertex of the inverted U-shaped portion is set at a position at least higher than the maximum liquid surface level of the reagent in the processing container.

According to such a structure, the spontaneous discharge of the reagent can be suppressed by the peculiar form of the sample flow channel without the need for the reagent valve. For example, when a certain amount of reagent was introduced, the other end of the reagent flow path was pulled upward from the liquid surface of the reagent, and air was slightly sucked (until the air crossed the inverted U-shaped apex). After that, if the operation of the air pump is stopped and then the inside of the processing container is returned to atmospheric pressure, due to the action unique to the inverted U-shape of the reagent flow path, that portion is used as a watershed to prevent the reagent from flowing out, and the desired amount of reagent is Can be held in the processing container. Then, if the air pump is rotated in the reverse direction after the processing is completed, the reagent can be discharged.

The atmosphere in the processing container can be opened, for example, by temporarily disconnecting the air pump from the processing container, or by providing an opening for opening the atmosphere at a predetermined position of the cylinder. If the air pump is individually arranged for each processing container, the reagent may be held in the processing container by maintaining the stopped state of the air pump.

In a preferred aspect of the present invention, a filter for placing a sample of biological tissue is provided in the processing container.
That is, the problem that the sample is placed on the filter in the processing container and the sample blocks the one end opening of the reagent channel when the reagent is discharged is solved. Of course, the filter does not hinder the flow of the reagent, and is made of a porous material, a mesh material, or the like.

Further, in a preferred aspect of the present invention, a reagent container table provided with a plurality of reagent containers, a means for selecting a reagent container to be used among the plurality of reagent containers, and the other end of the reagent flow path. Means for advancing into the reagent container to be used. That is, any one of the plurality of reagent containers provided in the reagent container table is selectively used, and the other end of the reagent channel is guided into the reagent of the selected reagent container.

Further, in a preferred aspect of the present invention, it includes a processing container table having a plurality of processing containers, and a means for selecting a processing container into which a reagent is introduced from among the plurality of processing containers. That is, a sample is stored in each of a plurality of processing containers provided in the processing container table, and each processing container is sequentially selected to introduce and discharge a reagent.

[0018]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT A preferred embodiment of the present invention will be described below with reference to the drawings.

FIG. 1 shows the essential structure of a biological tissue processing apparatus according to the present invention. In FIG. 1, the processing container 20 has a cylindrical shape as a whole, and a mesh filter 24 for mounting a sample 22 is arranged below the processing container 20. This processing container 20 has, for example, an inner diameter of 5 mm and its height is, for example, 5 mm. In addition, each member is
It is composed of a material that does not adversely affect the sample and is stable to the reagent. A tube 26 as an inverted U-shaped reagent channel as shown in FIG. 1 is connected to the lower portion of the processing container 20. Specifically, one end of the tube 26 is connected to the lower portion of the processing container 20, and the tube 26
Extends upward from one end thereof to form an inverted U-shaped portion, and the other end extends to a position lower than the floor surface of the processing container. The bottom surface of the processing container 20 is slightly inclined to the one end side of the tube 26. The apex of the inverted U-shaped portion of the tube 26 is set at a position at least higher than the maximum liquid surface level of the reagent in the processing container 20. The unique shape of the tube 26 prevents spontaneous outflow of reagents from the process vessel 20 without the need for reagent valves.

An air pump 28 composed of, for example, a syringe is connected to the upper portion of the processing container 20. That is, the inside of the air pump 28 and the inside of the processing container 20 communicate with each other via the connection hole 30 formed in the upper portion of the processing container 20, and the pressure inside the processing container 20 is increased or decreased by driving the piston 32 of the air pump 28. can do. The air pump 28 is detachable from the connection hole 30,
The air pump 28 can be connected to the connection hole 30 as needed. The piston 32 is driven, for example, artificially, or automatically using a motor or the like (not shown).

In order to store the sample 22 in the processing container 20, it is desirable to provide a lid structure on the upper part of the processing container 20. That is, a lid is provided so as to be openable and closable with respect to the main body of the processing container 20, the lid is opened, the sample 22 is put inside, and then the lid is closed. In this case, it is desirable to provide the lid with a sealing member such as an O-ring, which is necessary to maintain the airtight space inside the processing container 20.

The above-mentioned filter 24 is composed of a sheet made of mesh-shaped polypropylene, for example, and the sample 22 is discharged by the filter 24 when the reagent is discharged.
It is possible to avoid the problem that the tube closes the one end opening of the tube 26. When the sample 22 moves unnecessarily in the processing container 20, a second filter may be provided on the upper side of the sample 22 to sandwich the sample with the two filters to prevent the movement. it can. Such a second filter can be formed, for example, inside the lid described above.

FIG. 2 shows a series of steps for treating a biological tissue with a reagent using such a treatment container 20. As shown in FIG. 2A, the other end of the tube 26 is inserted into the reagent container 36 with the piston 32 being completely pushed into the cylinder 34. In this case, since the liquid level drops as the reagent is sucked, the other end of the tube 26 needs to be extended to a position below which the liquid level drop of the reagent is not affected.

When the piston 32 is pulled up as shown in FIG. 2B from the state shown in FIG. 2A, the air pump 2
By the action of 8, the air pressure in the processing container 20 decreases,
Along with this, the reagent in the reagent container 36 is introduced into the processing container 20 via the tube 26. Since the introduction of such a reagent is performed statically, the reagent does not scatter or the sample does not jump outside when the reagent is introduced.

When a certain amount of the reagent is introduced into the processing container 20, the reagent container 36 is removed from the other end of the tube 26 as shown in FIG. The other end of the tube 26 is pulled upward from the surface,
Suction of air instead of the reagent is subsequently performed. As a result, air is taken into the tube 26. FIG.
As shown in (D), the air pump 28 is removed from the connection hole 30 of the processing container 20 when the air is sucked to a position beyond the inverted U-shaped portion of the tube 26. As a result, the inside of the processing container 20 is returned to the atmospheric pressure.
Due to the inverted U-shape of 6, the reagent introduced inside does not flow out. That is, the outflow prevention function can be realized by the form of the tube 26 itself without requiring a reagent valve or the like. If a valve such as a valve is provided,
Although it is necessary to provide a drive mechanism for the operation and operation, according to the present invention, the need is eliminated, and the mechanism of the device can be extremely simplified. In the state shown in FIG. 2D, the reagent treatment is performed by leaving it for a certain period of time. If necessary, heating may be performed in this state. Further, by applying a predetermined vibration to the processing container 20, the efficiency of the reagent processing can be improved. Such vibration can be performed by moving the piston up and down while the air pump 28 is connected, for example.

After the elapse of a predetermined time, as shown in FIG. 2 (E), the air pump 28 is again connected to the connection hole 30 and the piston 32 is further pushed, so that the air pressure inside the processing container 20 increases. The reagent introduced inside according to this is discharged to the outside through the tube 26. The discharged reagent may be sent to the waste tank as it is for processing, but if possible, it may be discharged back into the reagent container 36 for reuse. FIG. 2 (F) shows a state in which the reagent has been completely discharged. From the state shown in (F), a series of steps are executed again for the next reagent.

FIG. 3 shows another embodiment of the air pump 28. In this embodiment, an air hole 40 is formed at a predetermined position above the cylinder 34, and the piston 3
The inside of the air pump 28 is automatically returned to the atmospheric pressure when the tip end surface of the valve 2 is lifted upward and exceeds the air hole 40. That is, by forming such an air hole 40 at an appropriate position, the detaching operation of the air pump 28 shown in FIG. 2D can be omitted. Of course, the position of the air hole 40 is provided at a position corresponding to the amount of the reagent to be introduced into the processing container 20. A plurality of openable / closable air holes may be formed along the movement direction of the piston, and any one of the air holes may be selectively used according to the reagent amount.

FIG. 4 shows the overall structure of the biological tissue processing apparatus according to the present invention. In this device, each mechanism is automated.

In FIG. 4, the reagent container table 50 holds a plurality of reagent containers 36 upright along the circumferential direction thereof. The reagent container table 50 is rotated by a rotation drive mechanism (not shown).
Further, this apparatus has a push-up mechanism for pushing the selected reagent container 36 upward. Specifically, by pushing up the lower surface of the reagent container 36 with the push-up rod 52, the tube 26 The other end of the is inserted into the reagent container 36. Of course, although the reagent container 36 is moved upward in this embodiment, conversely, the other end of the tube 26 may be moved into the reagent container 36 by moving the processing container 20 downward.

On the other hand, the processing container table 54 holds a plurality of processing containers 20 upright along the circumferential method. The processing container table 54 is rotated by a rotation drive mechanism (not shown), and one of the processing containers 20 is selected to introduce the reagent and discharge the reagent. Specifically, the processing container 20 into which the reagent is introduced is positioned at the position closest to the reagent container table 50, and in that state, the push-up rod 5 is placed as described above.
The reagent is introduced by moving the reagent container 36 selected by 2 upward. An air pump 28 is arranged above the processing container 20 into which the reagent is introduced. Specifically, the air pump 28 is held by a drive mechanism (not shown), and as shown in FIG. 2, the air pump 28 is connected to the connection hole 30 of the processing container 20 and detached as necessary. .

In the apparatus shown in FIG. 4, a plurality of samples can be treated in parallel with each other,
That is, while one reagent is introduced into the processing container 20 and the reagent processing is being performed, the predetermined reagent can be sequentially introduced into the other processing container to perform the reagent processing. The rotation speed of the processing container table 54 is determined corresponding to the time required for such reagent processing. In addition, in the apparatus shown in FIG. 4, an air pump 28 can be provided for each processing container 20.

[0032]

As described above, according to the present invention,
It is possible to provide an automated biological tissue processing apparatus capable of smoothly aspirating and discharging a reagent with a simple configuration and efficiently performing a series of reagent processing.

[Brief description of drawings]

FIG. 1 is a diagram showing a main configuration of a biological tissue processing apparatus according to the present invention.

FIG. 2 is a process diagram showing an actual treatment by the biological tissue treating apparatus according to the present invention.

FIG. 3 is a view showing another embodiment of the air pump.

FIG. 4 is a diagram showing an overall configuration of a biological tissue processing apparatus according to the present invention.

FIG. 5 is a diagram showing the concept of a conventional biological tissue processing apparatus.

[Explanation of symbols]

20 processing container, 22 sample, 24 filter, 26
Tube, 28 air pump.

Claims (6)

[Claims] 1. A processing container for containing a sample of biological tissue, a reagent container containing a reagent, and a reagent channel in which one end is connected to a lower portion of the processing container and the other end is introduced into the reagent container. And an air pump connected to an upper portion of the processing container, wherein the air pump is operated to introduce a reagent into the processing container through the reagent flow path to process biological tissue. Biological tissue processing device. 2. The apparatus according to claim 1, wherein the reagent channel extends upward from the one end, bends in an inverted U shape in the middle, and then extends downward to form the other end. The apparatus for treating biological tissue, wherein the other end is formed at a position lower than at least the one end. 3. The apparatus according to claim 2, wherein the apex of the inverted U-shaped portion is set at a position at least higher than the maximum liquid level of the reagent in the processing container. apparatus. 4. The apparatus for treating biological tissue according to claim 1, wherein a filter for placing a sample of biological tissue is provided in the treatment container. 5. The apparatus according to claim 1, wherein a reagent container table having a plurality of reagent containers, a means for selecting a reagent container to be used from the plurality of reagent containers, and the other end of the reagent flow path. And a means for advancing the above into the reagent container to be used. 6. The apparatus according to claim 1, further comprising: a processing container table having a plurality of processing containers; and a unit for selecting a processing container into which a reagent is introduced from among the plurality of processing containers. And biological tissue processing equipment.