JPH09304251A - Apparatus and container for treating biological tissue - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a mechanism efficiently removing a reagent after the completion of treatment by simple constitution in a biological tissue treatment apparatus reacting a reagent with a sample of biological tissue to treat the sample. SOLUTION: A discharge port 126a provided to the tip of a discharge pipe 126 of a treatment container 12 has a diameter as small as a capillary tube in dimension. Therefore, when a reagent 30 is injected into the treatment container 16, the reagent 30 is not discharged from the discharge port 126a by the action of the surface tension of the reagent until the reagent 30 becomes a certain amount or more and a predetermined amt. of the reagent 30 is held within the treatment container 12. By operating a suction pump 22 after the completion of reagent treatment, the balance between the liquid pressure at the discharge port 126a held heretofore and the surface tension of the reagent 30 is collapsed and the reagent 30 is discharged from the discharge port 126a. By this constitution, even if a mechanical mechanism such as a valve is not provided to the discharge port 126a, the holding and discharging of the reagent can be controlled.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an apparatus for treating biological tissue for treating a sample of biological tissue with a reagent, and more particularly to an apparatus used for whole mount treatment for treating biological tissue as it is without slicing.

[0002]

2. Description of the Related Art In immunostaining processing and ISH (in situ hybridization) processing, which are performed in fields such as genetic diagnosis, in addition to the processing of thinly slicing a sample of biological tissue and pasting it on a slide glass, There is a method of treating a sample as it is with a reagent. This method is called whole mount processing, and is often used for processing minute samples such as chicken embryos.

FIG. 8 is a diagram for explaining the procedure of a conventional hole mount process. Conventional hole mount processing is 2
A 4-well or 96-well microplate 200 is used, and the sample 20 is first placed in each container 202 of the microplate 200.
4 is accommodated, and the reagent 206 is dispensed into each accommodation portion 202 using the pipette 208 (FIG. 8A). Then reagent 2
In order to promote the reaction between 06 and the sample 204, an operation of placing the microplate 200 on an agitator and agitating it, or an operation of putting the microplate 200 in a constant temperature bath and keeping it at an appropriate temperature are performed. When the treatment with the reagent 206 is completed in this way, the tip of the fine glass tube 210 connected to the pump is inserted into the container 202, and the reagent 206 is inserted into the container 202.
It is removed by suction (FIG. 8 (b)). Immunostaining and IS
Since a plurality of reagents are usually used in H treatment and the like, the procedures of reagent dispensing, reaction, and reagent removal described above are repeated a plurality of times.

[0004]

In the conventional hole mount process, in order to suck all the reagent inside the container 202, it is necessary to bring the tip of the glass tube 210 close to the bottom of the container 202. Such delicate control is relatively difficult to automate, and a large-scale control mechanism is required if it is intentionally automated. In particular, since the microplate 200 is provided with a large number of accommodating portions 202,
To remove these reagents efficiently, the glass tube 2
It is necessary to take measures such as providing a plurality of 10 and performing suction in parallel, and the device configuration tends to increase in scale.

Further, in the conventional whole mount process, when the reagent is removed, the sample 204 is not sucked into the glass tube 210 or the tip of the glass tube 210 is not clogged with the sample 204, for example. It was necessary to hold the sample 204 with tweezers. However, it is extremely difficult to automate the delicate work of holding down the minute sample 204, so
This work had to be done manually, and thus the work efficiency was extremely poor.

[0006] In research and inspection in the field of gene diagnosis,
Since it is necessary to prepare many specimens, labor saving of specimen preparation work is required, and an apparatus capable of efficiently removing a reagent with a simple configuration is desired.

The present invention has been made in order to solve such a problem, and it is possible to efficiently carry out the reagent removal process with a simple structure, and thus to efficiently prepare a large amount of specimens. An object of the present invention is to provide a treatment device for biological tissue that can be used.

[0008]

In order to achieve the above-mentioned object, a biological tissue processing apparatus according to the present invention arranges a sample of biological tissue and injects a reagent into a processing container, and uses the sample as a reagent. The biological tissue processing apparatus for processing is characterized in that a capillary-shaped reagent discharge port is provided at the bottom of the processing container.

In this configuration, since the reagent discharge port provided at the bottom of the processing container is a capillary-sized opening, when the reagent is injected into the processing container, the liquid level of the reagent is the surface of the reagent at the reagent discharge port. The reagent is not discharged from the reagent discharge port until the tension and the liquid pressure at the position are at a level that balances them. Conversely speaking, the capillary-sized reagent discharge port means that the surface tension and the liquid pressure (that is, the reagent acts on the reagent) at the reagent discharge port when the reagent is accommodated in the processing container up to a predetermined liquid level. Gravity) means an opening of a size comparable to that of gravity. With such a mechanism, the sample can be processed by the reagent held in the processing container.

After completion of the reagent treatment, the reagent can be discharged from the reagent discharge port by sucking from the reagent discharge port or applying pressure to the reagent from the upper part of the processing container.

According to this structure, the reagent in the processing container can be efficiently removed with a relatively simple structure. In particular, this configuration has an advantage that the holding and discharging of the reagent can be controlled without providing a special mechanism such as a valve at the reagent discharging port.

In this configuration, the position at which the reagent discharge port is provided is not limited to the strict bottom surface of the processing container. For example, the reagent in the processing container may be located at the bottom of the side surface of the processing container. It suffices if it is a position through which almost all can be discharged. Further, a configuration is also provided in which a hole is provided in the bottom surface or the lower side surface of the processing container, a tube having a diameter of a capillary tube is extended downwardly or laterally from the hole, and the reagent is discharged from the opening at the tip of the tube. Included in the range.

Further, according to one aspect of the present invention, when the treatment with the reagent held in the treatment container is completed, the next reagent is injected into the treatment container, and the liquid pressure is increased by the injection to increase the treatment. The reagent held in the container is discharged from the reagent discharge port and replaced with the next reagent. According to this configuration, since the previous reagent is automatically discharged by the next injection of the reagent, it is possible to discharge the reagent without providing a reagent discharging mechanism.

In another aspect of the present invention, the reagent outlet is connected to a suction means, and when the reagent is removed from the processing container, the suction means is operated to remove the reagent from the reagent outlet. Suction. In this configuration, by operating the suction means, the atmospheric pressure near the reagent discharge port is reduced, and the reagent is rapidly sucked and discharged from the reagent discharge port.

Further, in another aspect of the present invention, a pressurizing means for applying a pressure is provided on the upper surface of the reagent contained in the processing container, and the pressurizing means is used for removing the reagent from the inside of the processing container. By applying pressure to the reagent from above, the reagent in the processing container is discharged from the reagent discharge port.
According to this configuration, the reagent is rapidly discharged from the reagent discharge port by applying pressure to the reagent from above by the pressurizing means. In a biological tissue processing apparatus, a lid mechanism for sealing the upper side of the processing container may be provided, and during processing of the reagent, the lid may cover the processing container to reduce evaporation of the reagent. The lid mechanism may be provided with a pressing means. According to this configuration, since the lid mechanism is provided with the pressurizing means, the device configuration can be simplified.

The biological tissue processing apparatus according to the present invention is a container rack to which a plurality of the processing containers can be detachably mounted, and has a waste liquid pipe connected to the reagent outlet of each of the processing containers. A container rack is provided, and the reagent in each processing container is discharged through the waste liquid pipe.

In this structure, by mounting a plurality of processing containers on the container rack, it is possible to handle a large number of samples as in the conventional microplate. Then, the reagent discharged from each processing container is discharged from a waste liquid pipe provided in the container rack. This configuration is extremely advantageous in terms of cost compared with the conventional microplates in terms of being disposable. In other words, in containers used for inspections, there is a demand to dispose of containers that are used once and may be contaminated in order to save the labor of cleaning, but with conventional microplates, only a part of the container is used. Even if it did not exist, the microplate would have to be discarded, resulting in a large cost burden. On the other hand, in this configuration, since each processing container can be disposable, there is no waste.

Further, the biological tissue processing apparatus according to the present invention comprises suction / blockage prevention means for preventing the sample from being sucked into the reagent discharge port and the reagent discharge port from being blocked by the sample in the processing container. It is characterized by being provided. As the suction / blockage prevention means, for example, a mesh member having a finer mesh than the sample can be used. Such a suction
By providing the blockage prevention means, it is possible to prevent the sample from being discharged together with the reagent at the time of discharging the reagent and to prevent the reagent discharge port from being blocked by the sample, so that the reagent can be discharged smoothly.

The biological tissue processing container according to the present invention is characterized in that it has a capillary-shaped reagent discharge port at the bottom. This processing container can hold and discharge the reagent without using a special mechanism such as a valve. If this processing container is provided with a suction / blockage prevention unit that prevents the sample from being sucked into the reagent discharge port and from blocking the reagent discharge port by the sample, the reagent can be discharged smoothly.

[0020]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Preferred embodiments of the present invention will be described below with reference to the drawings.

FIG. 1 is a perspective view schematically showing the overall structure of the biological tissue processing apparatus according to the present invention.

The biological tissue processing apparatus has a container rack 10, and a plurality of processing containers 12 are provided in the container rack 10.
Is attached. More specifically, the processing container 12 is mounted on the container rack 10 via the holder 14. Holder 14
Is a plate-shaped member and has a predetermined number of holes into which the processing container 12 is inserted. The container rack 10 is placed on the heater 16 and the shaker 18. Heater 16
Is a device for controlling the temperature during reagent processing, and the shaker 18 is a device for applying vibration to the reagent to promote the reaction.

The dispensing head 26 is constructed so as to be movable in three axial directions, and when a sample is set in each processing container 12, an appropriate amount of a necessary reagent is sucked up from the reagent bottle table 28 to be stored in each processing container 12. Dispense.

After dispensing the reagent, the lid mechanism 24 moves to cover and seal the upper surface of each processing container 12 of the container rack 10.
The lid mechanism 24 is configured to reduce the evaporation of the reagent. Then, in this state, the temperature of the reagent in each processing container 12 is controlled to an appropriate temperature by the action of the heater 16, and the shaker 18 is further operated as necessary to promote the reaction between the reagent and the sample.

When the treatment with the reagent is completed, the lid mechanism 24
Rises and the suction pump 22 operates. As will be described later, the container rack 10 is provided with a waste liquid pipe connected to the discharge port of each processing container 12, and the suction pump 22
This is a container rack 1 via a Teflon tube 20.
It is connected to 0 waste liquid pipe. Therefore, the suction pump 22
When is activated, the reagent is discharged from the discharge port of the processing container 12, and the discharged reagent is discharged to a waste liquid bottle (not shown) through the tube 20 and the suction pump 22.

The overall configuration of this embodiment has been briefly described above. Next, the processing container 1 according to the present embodiment
The structure of No. 2 and the mechanism for discharging the reagent from the processing container 12 will be described with reference to FIGS. 2 and 3.

As shown in FIG. 2, the processing container 12 of the present embodiment has a truncated cone-shaped cup portion 120 and a cup portion 12.
0 and a discharge pipe 126 extending from the bottom surface. The upper portion of the cup portion 120 has an outer edge 122 protruding outward so as to be hooked by the holder 14. The size of the cup portion 120 is related to the size of the holder 14 and the container rack 10, and for example, the diameter of the upper opening is 14 mm, the diameter of the bottom surface 124 is 10 mm, and the depth is 12 m.
m.

A hole having a capillary size (for example, about 1 mm) is provided on the bottom surface 124 of the cup portion 120, and a discharge pipe 126 having a length of 10 mm and having the same inner diameter with respect to the hole.
Is connected. Further, the lower end of the discharge pipe 126 is a discharge port 126a having the same capillary size. Such a processing container 12 can be formed by resin molding, for example.

In FIG. 3, the processing container 12 is provided with a container rack 1.
It is the figure which showed the state mounted | worn with 0. As shown in FIG. 3, the container rack 10 has holes 1 for accommodating the processing containers 12.
00 is drilled in a predetermined number. The container rack 10 is made of, for example, aluminum. In addition, in the holder 14,
A hole into which the processing container 12 is inserted is provided. The diameter of this hole is large enough to catch the outer edge of the processing container 12. The holder 14 is provided to remove a plurality of processing containers 12 from the container rack 10 at once after all the reagent processing is completed.

The processing container 12 is mounted on the container rack 10 manually. In this case, the operator stacks the holder 14 on the container rack 10 and then inserts the processing container 12 into the container rack 10 through the hole of the holder 14.
A seal member 104 made of rubber is provided at the lower end of the hole 100 of the container rack 10, and the discharge pipe 126 of the processing container 12 is inserted and fixed in this seal member. As a result, the discharge port at the tip of the discharge pipe 126 projects into the waste liquid pipe 102 provided in the container rack 10. The waste liquid pipe 102 has a predetermined airtightness due to the action of the seal member 104.

When the mounting of the processing container 12 is completed, the sample 32 is set in each processing container 12, and the dispensing head 26 is operated to dispense a predetermined reagent into each processing container 12.

In the processing container 12 of this embodiment, the discharge pipe 1
Since the discharge port 126a at the tip of 26 is an opening having a diameter of a capillary tube, when the reagent 30 is injected into the processing container 12, the surface tension of the reagent 30 causes the amount of the reagent 30 to reach a certain amount or more (that is, the reagent). The reagent 30 is not discharged from the discharge port 126a until the liquid surface of 30 becomes higher than a certain height. That is, while the liquid pressure due to the weight of the reagent 30 and the surface tension of the reagent 30 are balanced at the discharge port 126a, the reagent 30 is not discharged from the discharge port 126a and is held in the processing container 12. Then, the reagent 30 is discharged from the discharge port 126a only when the injection amount exceeds the limit amount and the surface tension cannot withstand the liquid pressure. By this mechanism, a predetermined amount of reagent 30 is held in the processing container 12.

When the treatment of the sample 32 with the reagent 30 is completed, the suction pump 22 is activated and the container rack 10
The waste liquid pipe 102 is pulled. As a result, the balance between the liquid pressure and the surface tension maintained until then is broken, and the reagent 30 is sucked out to the waste liquid pipe 102 from the discharge port 126a. The discharged reagent 30 is discharged into the waste liquid bottle via the tube 20 and the suction pump 22. And after a certain period of time,
When all the reagents in each processing container 12 are discharged, the suction pump 22 is stopped. Therefore, when another reagent is injected into the processing container 12 thereafter, a predetermined amount of the reagent is held in the processing container 12 due to the effect of the surface tension.

As described above, according to the present embodiment, by making the discharge port 126a of the processing container 12 an opening having a capillary size, it is possible to hold and discharge the reagent without providing a mechanical mechanism such as a valve. It can be switched in conjunction with the operation of the suction pump 22. Therefore, according to the present embodiment, the reagent in each processing container 12 can be automatically and efficiently removed with a relatively simple configuration.

In the configuration of the above embodiment, the waste liquid pipe 1
Although the reagent 30 in the processing container 12 was sucked out by pulling 02 with the suction pump 22, there are the following methods for discharging the reagent.

One is a method of applying pressure from above the reagent 30 in the processing container 12. FIG. 4 is a schematic diagram showing a configuration for realizing this method. In the configuration of FIG. 4, a pipe 24a is provided inside the lid mechanism 24 that hermetically seals the processing container 12, and air is supplied to each processing container 12 from this piping 24a by an air pump (not shown), so that the upper surface of the reagent 30 is removed. It can be pressurized. In this configuration, after the reagent 30 is dispensed into each processing container 12, the lid mechanism 24 is moved to cover the processing container 12, and the treatment with the reagent 30 is performed. Then, when the processing is completed, each processing container 12 is pressurized by the air pump with the lid kept. As a result, the reagent 30 passes through the discharge pipe 126 and is discharged from the discharge port 126a to the waste liquid pipe 102. With this configuration, holding and discharging of the reagent in the processing container 12 can be switched by the operation of the air pump.

In this configuration, the reagent 30 in the processing container 12
Although the means for pressurizing the lid mechanism 24 is built in the lid mechanism 24, such a pressing means may be provided separately from the lid mechanism 24.

Another method for discharging the reagent is to further inject a liquid into the processing container 12 holding the reagent 30 so that the amount of the liquid in the processing container 12 exceeds the limit supported by the surface tension. It is a method of increasing to. That is, in this method, for example, the dispensing head 26
Then, the liquid is further injected into each processing container 12. As a result, the liquid pressure at the discharge port 126a increases to overcome the surface tension, and the reagent 30 in the processing container 12 is discharged from the discharge port 126a. According to this method, the holding / discharging of the reagent can be controlled without providing a special means for suction or pressurization. The processing container 12 is used to discharge the reagent.
If the liquid to be injected into is the reagent used for the next reagent treatment, the next reagent can be injected at the same time when the treated reagent is discharged, and the treatment efficiency is improved.

Next, a modification of the processing container 12 will be described.

FIG. 5 shows an example in which the processing container 12 is provided with a filter 128. In this configuration, the bottom surface 1 of the processing container 12
A filter 128 is provided slightly above 24. As the filter 128, for example, a mesh having an opening of about 0.4 mm square can be used. By providing the filter 128, it is possible to prevent the sample 32 from being sucked into the discharge tube 126 and the sample 32 from blocking the inlet of the discharge tube 126 when the reagent is discharged. As the filter 128, a filter other than a mesh may be used as long as it allows a reagent to pass but does not allow a sample to pass. Also, for example, the bottom surface 12
The filter 128 may be provided in the opening portion of No. 4.

FIG. 6 shows an example in which a discharge pipe 130 is provided at the lower end of the side surface of the processing container 12. In the example of FIG. 2 described above, the discharge pipe is provided on the bottom surface of the processing container 12, but the disposition position of the discharge pipe is not limited to this, and it may be any position where all the reagents in the processing container 12 can be discharged. Good. Therefore, even when the discharge pipe 130 is provided at the lower end portion of the side surface of the processing container 12 as shown in FIG. 6, the effect of this embodiment can be obtained.

FIG. 7 shows another example of the processing container 12. In FIG. 7, the discharge port 132 is only provided on the bottom surface of the processing container 12, and the discharge pipe as in the example of FIG. 2 is not provided. Even in the case of such a structure, the outlet 132
If the diameter of is the capillary dimension, it is possible to obtain the same effect as the above-mentioned example.

In the embodiment described above, since each processing container 12 can be disposable, all the microplates are discarded although only some holes are used as in the conventional microplate. There is no waste of having to do it.

[Brief description of drawings]

FIG. 1 is a perspective view schematically showing an overall configuration of a biological tissue processing apparatus.

FIG. 2 is a perspective view showing the structure of a processing container used in the biological tissue processing apparatus according to the present invention.

FIG. 3 is a cross-sectional view showing a state in which a processing container is mounted on a container rack.

FIG. 4 is a cross-sectional view showing a configuration example in which a reagent in a processing container is pressurized to discharge the reagent.

FIG. 5 is a diagram showing an example in which a filter is provided in a processing container.

FIG. 6 is a diagram showing an example in which a discharge pipe is provided at a lower end portion of a side surface of a processing container.

FIG. 7 is a diagram showing another example of the configuration of the processing container.

FIG. 8 is an explanatory diagram showing a procedure of reagent processing in a conventional biological tissue processing apparatus.

[Explanation of symbols]

10 container racks, 12 processing containers, 14 holders, 1
6 heater, 18 shaker, 20 tube, 22 suction pump, 24 lid mechanism, 26 dispensing head, 28 reagent bottle table, 30 reagent, 32 sample, 102 waste liquid pipe, 104 sealing member, 126 discharge pipe, 126a
Vent.

Claims (10)

[Claims] 1. A biological tissue processing apparatus for arranging a sample of biological tissue and injecting a reagent into a processing container to treat the sample with a reagent, wherein a capillary-shaped reagent discharge port is provided at the bottom of the processing container. A biological tissue processing apparatus characterized by the above. 2. The biological tissue processing apparatus according to claim 1, wherein when the treatment with the reagent held in the treatment container is completed, the next reagent is injected into the treatment container, and the fluid pressure due to the injection is increased. An apparatus for treating a biological tissue, wherein the reagent retained in the treatment container due to the increase is discharged from the reagent discharge port and replaced with the next reagent. 3. The biological tissue processing apparatus according to claim 1, wherein the reagent discharge port is connected to a suction means, and when the reagent is removed from the processing container, the suction means is operated to discharge the reagent. An apparatus for treating biological tissue, characterized in that a reagent is sucked from an outlet. 4. The biological tissue processing apparatus according to claim 1, further comprising a pressurizing unit that applies a pressure to the upper surface of the reagent contained in the processing container, when the reagent is removed from the inside of the processing container. Is an apparatus for treating biological tissue, characterized in that the reagent in the treatment container is discharged from the reagent discharge port by applying pressure to the reagent from above by the pressurizing means. 5. The biological tissue processing apparatus according to claim 4, wherein the pressurizing unit is provided in a lid mechanism used to seal the upper surface of the processing container. 6. The biological tissue processing apparatus according to claim 1, wherein the processing container is a container rack to which a plurality of the processing containers can be detachably mounted, and a waste liquid pipe connected to a reagent outlet of each of the processing containers. A treatment apparatus for biological tissue, comprising a container rack having the same, and discharging the reagent in each treatment container through the waste liquid pipe. 7. The biological tissue processing apparatus according to claim 1, wherein suction of the sample into the reagent outlet and blocking of the reagent outlet due to the sample are prevented in the processing container. An apparatus for treating biological tissue, characterized in that it is provided with means for preventing suction and blockage. 8. The biological tissue processing apparatus according to claim 7, wherein the suction / closure prevention unit is a mesh member that covers at least the reagent discharge port and has a finer mesh than the size of the sample. Biological tissue processing equipment. 9. A biological tissue processing container for holding a biological tissue sample and a reagent, wherein the biological tissue processing container has a capillary-sized reagent discharge port at the bottom. 10. The treatment container for biological tissue according to claim 9, wherein the treatment container prevents suction of the sample into the reagent discharge port and blockage of the reagent discharge port by the sample. A treatment container for biological tissue, comprising a means.