JPH0321475Y2 - - Google Patents

Description

[Detailed description of the invention] (Industrial application field) The chemical liquid shaking mechanism of the embedding device for preparing electron microscope specimens according to this invention is used in the medical or natural science fields to prepare specimens to be observed with an electron microscope. In order to do this, we have installed an embedding device that fixes samples taken from living organisms and embeds them in resin automatically and with a small amount of chemical solution. This improves familiarity.

(Prior art) In the field of medicine or natural science, when a collected sample is observed using a microscope, the sample is treated with a chemical solution, a resin is infiltrated into the sample, and the resin is solidified together with the sample. The embedding process is performed to The sample that has undergone the embedding process is sliced and attached to a glass slide to prepare a specimen for microscopic observation.

Conventionally, as an embedding device that can automatically perform the above-mentioned sample embedding process, a plurality of bottomed cylindrical units (for example, 20 units) that store processing chemicals and resin are used. A structure is used in which containers are arranged on the same arc, and embedding baskets containing samples are sequentially immersed in the containers to embed the samples. The embedding cage is suspended from the outer periphery of the lower surface of a disk whose center part is fixed to a rotary lift shaft provided at the center of the plurality of containers, and the disk is moved up and down and rotated intermittently. , so that it can be immersed in multiple containers in sequence.

(Problems to be Solved by the Invention) However, in the conventional embedding device for preparing an electron microscope specimen as described above, the following disadvantages occur.

In other words, the sample to be embedded is small, about 0.5 to 1 m square, and is placed in an embedding cage with a diameter of about 1 cm and immersed in the processing solution in the container. This is when embedding is performed using 20 embedding baskets, which requires at least 20 to 30 c.c. In order to perform a good embedding process, the processing solution used once must be discarded.
The processing liquid contains quite expensive substances, and it is uneconomical to discard a relatively large amount of processing liquid, about 20 to 30 c.c., each time embedding treatment is performed.

Also, during the embedding process, the sample is first immersed in alcohol at a concentration of about 50%, then the concentration of this alcohol is gradually increased, and finally the sample is immersed in alcohol at a concentration of 100%. In the container
Alcohol with different concentrations of 50%, 70%, 80%, 95%, and 100% was stored and placed in order. However, since alcohol with concentrations of 50% and 100% is commercially available, it can be used as is, but alcohol with other concentrations must be adjusted each time, which is troublesome.

In order to solve this inconvenience, the inventor of the present invention
A chemical liquid selection mechanism that selectively communicates a plurality of liquid storage containers each storing a chemical liquid for embedding processing with a liquid feeding pipe equipped with a liquid feeding pump that can freely rotate in forward and reverse directions and prevents the flow of chemical liquid when stopped. , a plurality of embedding cages each having a short cylindrical shape and having a net-like plate in the opening at the lower end that does not allow the sample to be embedded to pass through are stacked vertically and held, and the liquid on the outer periphery of each embedding cage is The embedding cage holder is configured to maintain airtightness and has a lower connection port at the lower end for connecting the upper end of the liquid sending pipe, and an upper connection port for connecting one end of the overflow pipe leading to the waste liquid container at the upper end. and one of the plurality of connection ports provided on the waste liquid container and the chemical liquid selection mechanism.
He devised an embedding device for preparing specimens under an electron microscope that communicated with the individual through a drainage pipe, and filed a patent application on the same day (Japanese Patent Application No. 60-206578 (Japanese Patent Laid-open No. 62-67424)).

When embedding a sample using this embedding device for preparing electron microscope specimens, first stack multiple embedding baskets each containing a sample one on top of the other, and place the liquid in the vertical direction. These embedding baskets are combined into a circular tube shape for circulation and held by an embedding basket holder. Once the embedding cages have been held, the liquid medicine in the storage container is sent through the liquid feeding pipe into the embedding cages in the holder by operating the liquid feeding pump, and the drug solution is stored in each embedding cage. The sample is immersed in the chemical solution. During this immersion, the liquid feeding pump is stopped, and during this time the chemical liquid selection mechanism is switched to a state where the liquid feeding pipe and the liquid draining pipe are communicated with each other.

When the predetermined period of immersion is completed, the liquid feed pump is reversed to discharge the chemical solution used for the immersion treatment of the sample into the waste liquid container.

Once the waste liquid has been discharged, switch the chemical liquid selection mechanism again to connect the liquid feeding pipe to a liquid storage container storing another chemical liquid, rotate the liquid feeding pump in the normal direction, and place the liquid into the embedding cage again. Send the chemical solution.

Thereafter, by repeating the above-mentioned operations, the sample stored in the embedding cage is embedded.
When performing a process of immersion in alcohol from a low concentration to a higher concentration, first supply alcohol at a low concentration (50%) into the embedding cage, and then immerse the embedding cage in the embedding cage without draining this low concentration alcohol. Continuously supply high concentration (100%) alcohol so that the concentration of alcohol in the embedding cage gradually increases.

By the way, in the newly developed embedding device for preparing electron microscope specimens that is configured and operates in this way, unlike the case of conventional embedding devices, the embedding basket cannot be shaken in the chemical solution, and the embedding In order to mix the sample in the burial cage with the chemical solution, the chemical solution must be shaken.

The present invention provides a chemical solution shaking mechanism for an electron microscope specimen embedding device that can be incorporated into such a newly developed electron microscope specimen embedding device to easily shake a chemical solution.

(Means for Solving the Problem) The chemical liquid shaking mechanism of the embedding device for preparing an electron microscope specimen of the present invention consists of a liquid storage container that stores a chemical liquid for embedding processing, and a short cylindrical container that is embedded in a lower end opening. A lower connection port of an embedding cage holder that holds a plurality of embedding cages with mesh plates that do not allow the sample to pass through, stacked vertically and keeping the outer periphery of each embedding cage liquid-tight. A hose made of a flexible material is connected in the middle of the liquid feeding pipe connecting the two, and clamping blocks provided at the tips of a pair of rotating arms each rotating around an axis are positioned on opposite sides of the hose. a spring that rotates the rotary arm in a direction in which the clamping blocks move away from each other; a solenoid that rotates the rotary arm against the elasticity of the spring to bring the pair of clamping blocks closer together; In addition to providing a control circuit for intermittent energization,
A portion of the liquid feeding pipe closer to the liquid storage container than the hose can be freely closed.

(Function) The chemical liquid shaking mechanism of the embedding device for preparing an electron microscope specimen of the present invention configured as described above shakes the chemical liquid sent into the embedding cage, and mixes the sample and the chemical liquid in the embedding cage. To blend in, do the following.

Once the drug solution is sent into the embedding basket holder that holds the embedding basket, the flow path in the middle of the liquid feeding pipe connecting this holder and the liquid storage container is closed at the part closer to the liquid storage container than the hose, and the control is performed. The circuit energizes the solenoid intermittently.

By intermittent energization of this solenoid, the interval between the holding blocks fixed to the pair of rotating arms repeatedly expands and contracts.

That is, as the solenoid is energized, the upper pair of rotating arms rotate in opposite directions against the elasticity of the spring, and the gap between the clamping blocks fixed to the tips of each rotating arm narrows. , this clamping block crushes the hose made of flexible material. When the hose is crushed in this way and the internal volume of the hose is reduced, the chemical liquid present above the hose flows upward.

Next, when the power to the solenoid is cut off, the pair of rotating arms rotate in the opposite direction to the above case due to the elasticity of the spring, and the gap between the clamping blocks widens, increasing the internal volume of the hose. The chemical liquid present above flows downward.

Therefore, if the solenoid is energized intermittently, the chemical liquid present above the hose will be shaken to flow finely up and down.

(Example) Next, the present invention will be explained in more detail while explaining the illustrated embodiment.

FIG. 3 schematically shows the overall configuration of an embedding device for preparing an electron microscope specimen incorporating the chemical solution shaking mechanism of the present invention. First, to explain this embedding device, the inside of the drug solution stocker 1 is divided into three chambers 4, 5, 6 by partition walls 2, 3, and each chamber 4, 5, 6 has one or more embedding devices. liquid storage containers 7a, 7
b and 7c are stored. Among these, processing liquids such as glutaraldehyde and sodium acetate are refrigerated at a low temperature of about 4° C. in liquid storage containers 7a and 7a in chamber 5.
Processing liquids such as 50% alcohol and 100% alcohol are stored in chambers b and 7b at room temperature, and resin such as resin is stored at around 37°C in liquid storage container 7c in chamber 6. ing.

At the bottom of each liquid storage container 7a, 7b, 7c, the lower ends of liquid suction pipes 8, 8 are open, respectively, and the upper ends of each liquid suction pipe 8, 8 are opened at the sockets 10, 8, of the liquid medicine selection mechanism 9, respectively. It leads to 10. This drug selection mechanism 9
is composed of a plurality of sockets 10, 10 whose positions are fixed, and a movable insertion port 11 which can be fluid-tightly joined to each socket 10, 10.

A liquid supply pipe 12 whose lower end communicates with this insertion port 11
A liquid feed pump 13 and a shaking mechanism 14 of the present invention are provided in order from the insertion port 11 side, and the upper end of this liquid feed pipe 12 is connected to a lower connection port provided at the lower end of the embedding basket holder 15. is connected to. The liquid feeding pump 13, like a peristaltic pump, can freely rotate forward and reverse, and when stopped, liquid does not flow through the pump. That is, use one that has a structure that closes the conduit.

An air reservoir 16 is provided at the upper end of the embedding basket holder 15, and constitutes an escape area into which the drug solution in the embedding basket holder 15 flows when the shaking mechanism 14 is activated.

One end of the overflow pipe 17 is connected to the upper connection port of the embedding basket holder 15 provided above the air reservoir 16, and the other end of the overflow pipe 17 communicates with the waste liquid container 18. This waste liquid container 18 has a three-way valve 19 in the middle.
The intake pipe 20 is connected to one end of the intake pipe 20, and the other end of the intake pipe 20 is provided with an air filter 21. An exhaust pipe 23 whose one end is connected to the remaining connection port of the three-way valve 19 and has a vacuum pump 22 installed in the middle.
An activated carbon filter 24 is provided at the other end.
Further, a drain pipe 2 having one end connected to the waste liquid container 18
The other end of 5 is communicated with one of the plurality of sockets 10 provided in the chemical liquid selection mechanism 9.

As shown in FIGS. 1 and 2, the shaking mechanism 14 is configured to intermittently crush a hose 27 made of a flexible material provided in the middle of the liquid feed pipe 12 using a clamp device 26. The clamp device 26 is attached to a base 28
By intermittently energizing a solenoid 29 fixed to the upper surface of the hose 27 by a control circuit (not shown), a pair of clamping blocks 30, 30 provided to sandwich the hose 27 are moved toward and away from each other.

Approximately L-shaped rotating arms 31, 31, each having a holding block 30, 30 fixed to its tip, are rotatable about a vertical shaft 32, respectively. Connecting rod 3 whose rear end is connected to the tension rod 33 of the solenoid 29
The flange-shaped portion formed at the tip of 4 is opposed to the rear end edge of the rotary arms 31, 31, and when the solenoid 29 is energized, the tension rod 33 is
When moved to the right in the figure, each rotating arm 31, 3
1 is rotated in a direction to narrow the gap between the holding blocks 30, 30. Also, each rotating arm 3
Compression springs 36, 36 are provided between 1, 31 and a block 35 fixed to the base 28, and each rotating arm 3
1 and 31 are given elasticity to rotate in the direction in which the gap between the holding blocks 30 and 30 is widened.

When embedding a sample using an embedding device for preparing an electron microscope specimen having a shaking mechanism configured as described above, a plurality of embedding baskets containing samples are set in the embedding basket holder 15, and then By operating the liquid feeding pump 13 in the middle of the liquid feeding pipe 12, the medicinal liquid in either of the liquid storage containers 7a (or 7b, 7c) is embedded in the embedding basket holder 15 through the liquid feeding pipe 12. The sample contained in each embedding cage is immersed in this chemical solution. During this immersion, the liquid feed pump 13 is stopped, and during this time, the chemical liquid selection mechanism 9 is switched to a state where the liquid feed pipe 12 and the liquid drain pipe 25 are communicated with each other.

In addition, while the liquid pump 13 is stopped, the solenoid 29 constituting the shaking mechanism 14 is intermittently energized to shake the medicinal solution that has entered the embedding cage, thereby causing the drug solution stored in each embedding cage to be shaken. Improve the compatibility between the sample and the chemical solution. That is, when the solenoid 29 is energized, the pair of rotating arms 31, 31 are compressed by the compression spring 3.
The holding block 3 rotates against the elastic force of 6 and 36.
0 and 30 becomes narrower, the hose 27 made of flexible material is crushed and the internal volume of this hose is reduced, and the chemical liquid present above this hose 27 flows upward, causing the excess chemical liquid to flow upward. is stored in the air reservoir 16. Next, when the energization to the solenoid 29 is stopped, the rotating arms 31, 31 release the compression springs 36, 36.
The clamping blocks 30, 30 rotate due to the elasticity of the
The interval between the hoses 27 and 27 widens, the hose 27 restores itself due to its own elasticity, and its internal volume increases, and the chemical liquid present above the hose 27 flows downward. By carefully repeating this operation, the chemical solution in each embedding cage and the sample become more compatible.

When the predetermined period of immersion is completed while performing such shaking, the liquid feed pump 13 is reversed,
The chemical solution used for the sample immersion process is discharged into the waste solution container 18. At the time of this discharge, air is sent into the embedding basket holder 15 through the overflow pipe 17 by switching the three-way valve 19 so that the inside of the waste liquid container 18 and the air filter 21 are communicated.
To ensure that the chemical liquid discharge work is carried out smoothly.
The vacuum pump 22 attached to the waste liquid container 18 is
It is used to reduce the pressure inside the embedding cage holder 15 when the sample in the embedding cage is treated with glutaraldehyde. Exhaust gas discharged during operation of the vacuum pump 22 is purified by passing through an activated carbon filter 24.

After draining the waste liquid, switch the chemical liquid selection mechanism 9 again to connect the liquid feeding pipe 12 to the liquid storage container 7a (or 7b, 7c) storing another chemical liquid, and adjust the liquid feeding pump 13 to the correct position. Turn it around and send the drug solution into the embedding cage again.

Thereafter, by repeating the above-mentioned operations, the sample stored in the embedding cage is embedded.
The above-mentioned shaking ensures that the chemical solution enters the inside of the embedding cage, which improves the compatibility between the sample and the chemical solution compared to the case of conventional processing equipment, allowing for better embedding processing. .

(Effects of the invention) Since the chemical solution shaking mechanism of the embedding device for electron microscope specimen preparation of the present invention is configured and operates as described above, the amount of chemical solution used for sample embedding processing can be reduced, and operating costs can be reduced. By incorporating it into an embedding device that can reduce the cost, it is possible to efficiently shake the chemical liquid sent into the embedding cage to improve the compatibility between the sample and the chemical liquid.

[Brief explanation of the drawing]

Figures 1 and 2 show an embodiment of the chemical solution shaking mechanism of the present invention, where Figure 1 is a side view, Figure 2 is a partial cross-sectional plan view, and Figure 3 is a schematic diagram showing the overall configuration of the embedding device. It is. 1...Medical liquid stocker, 2, 3...Partition wall, 4,
5, 6...chamber, 7a, 7b, 7c...liquid storage container,
8...Liquid suction pipe, 9...Medical solution selection mechanism, 10...Socket, 11...Insertion port, 12...Liquid sending pipe, 13...
Liquid feeding pump, 14... Shaking mechanism, 15... Embedding basket holder, 16... Air reservoir, 17... Overflow pipe, 18... Waste liquid container, 19... Three-way valve, 20...
... Intake pipe, 21 ... Air filter, 22 ... Vacuum pump, 23 ... Exhaust pipe, 24 ... Activated carbon filter, 25 ... Drain pipe, 26 ... Clamp device, 2
7... Hose, 28... Base, 29... Solenoid, 30... Clamping block, 31... Rotating arm, 3
2... Vertical shaft, 33... Tension rod, 34... Continuous rod, 3
5...Block, 36...Compression spring.

Claims (1)

[Scope of utility model registration request] A liquid storage container storing a chemical solution for embedding processing and a plurality of embedding baskets each having a short cylindrical shape and having a net-like plate in the lower end opening that does not allow the sample to be embedded to pass through are stacked vertically, A hose made of flexible material is connected to the middle of the liquid feed pipe that connects the lower connection port of the embedding cage holder that maintains the liquid tightness around the outer periphery of each embedding cage, and each of the embedding cages is rotated around its axis. The clamping blocks provided at the tips of a pair of moving rotating arms are located on opposite sides of the hose, and the clamping blocks have a spring that tries to rotate the rotating arms in a direction away from each other, and the elasticity of this spring. A solenoid is provided that rotates the rotary arm against the pressure to bring the pair of clamping blocks closer together, and a control circuit that intermittently energizes the solenoid. A chemical liquid shaking mechanism for an embedding device for preparing electron microscope specimens with a portion close to the liquid storage container that can be closed freely.