JPH0440356A - Electrophoresis device - Google Patents

Abstract

PURPOSE:To enhance the sepn. performance of a migration sample and t automate the sepn. and refining of the migration sample by providing a bubble removing means which removes the bubbles existing in the buffer soln. in a migration chamber. CONSTITUTION:A buffer soln. is previously injected from an introducing hole 3 into the migration chamber 1 to form parallel flow. The migration sample is cracked to each of the respective components when the migration sample is injected from an injection pipe 4 into the chamber. These components are fraction collected from a fraction collection port via a fraction collection pipe 7. The bubbles float on the buffer soln. surface if the bubbles exist in the buffer soln. at this time. The bubbles are sucked through a pass filter 19 for gas, a filter retainer 21 and a suction pipe 23 into a vacuum pump 22 and are dis charged to the outside. Namely, the bubbles can be surely discharged even if the bubbles exist in the buffer soln. and, therefore, the sepn. performance of the migration sample is enhanced. In addition, the efficiency of the sepn. operation is enhanced as well as the bubbles can be removed even if a power source is not interrupted.

Description

[Detailed Description of the Invention] [Industrial Application Field] The present invention relates to an electrophoresis device that electrophoretically separates a sample into various components, and particularly relates to an electrophoresis device that electrophoretically separates an electrophoresis sample into various components, and particularly relates to an electrophoresis device that electrophoresis separates an electrophoresis sample into various components. Concerning what is suitable for.

[Prior Art] Various electrophoresis devices for separating electrophoresis samples have been proposed and put into practical use.

Although not shown in the general electrophoresis apparatus, a buffer solution is supplied to the electrophoresis tank from the injection port to form a parallel flow downward along the height direction within the electrophoresis tank, and the electrophoresis is controlled. Inject the electrophoresis sample into the tank from the top.

When a DC voltage is applied to the ten and - electrodes of the electrolytic cell, the electrophoresis sample migrates into various components due to the moving force based on the amount of charge of each electrophoresis sample and the difference in fluid resistance that each particle of the electrophoresis sample receives. To separate.

At that time, among the electrophoretically separated components, those that flow down the center of the electrophoresis tank are separated from the separation section of the electrophoresis tank to the collection section for each component, and those that have shifted from the center of the electrophoresis tank are added to the buffer solution. At the same time, it is taken in from the discharge part of the electrophoresis tank.

In addition, in a migration tank, if air bubbles are present in the buffer solution, air bubbles may enter the buffer solution when it is injected into the migration tank.
Alternatively, if air bubbles are generated in the buffer solution in the electrophoresis tank, the bubbles will cause turbulence in the parallel flow of the buffer solution and the electrophoresis sample in the electrophoresis tank, which will deteriorate the separation performance of the electrophoresis sample. needs to be removed.

In the prior art, no means for removing air bubbles is provided, and air bubbles are removed by an operator as described below. That is, first cut off the power supply for energizing the + and - electrodes, then remove the cap on the top of the migration tank, and then inject a liquid such as a buffer from the bottom of the migration tank to drain the liquid in the migration tank. The air bubbles are removed by raising the cap to the upper cap attachment position, and the air bubbles also rise accordingly. Note that the reason to turn off the power when removing the cap is to ensure safety so that when liquid is injected into the migration tank, the liquid in the migration tank will leak outside and the worker will not receive an electric shock from the spilled liquid. is being considered.

[Problems to be Solved by the Invention] By the way, the above-mentioned prior art does not have a means for removing air bubbles present in the buffer solution, so the removal is done by an operator. As mentioned above, since the power supply is temporarily shut off, the separation and purification of the electrophoresing sample must be stopped during that time, resulting in a problem that the efficiency of separation and purification is reduced accordingly. In addition, since air bubbles are removed after a certain amount of bubbles have accumulated in the electrophoresis tank, the bubbles may cause turbulence in the electrophoresis tank until they are removed, which may reduce the separation performance of the electrophoresis sample. .

In view of the above circumstances, an object of the present invention is to improve the separation performance of electrophoresis samples by being able to reliably and easily remove bubbles even if they exist in the buffer solution in the electrophoresis tank without cutting off the power supply. An object of the present invention is to provide an electrophoresis device that can increase separation efficiency as well as increase separation efficiency.

[Means for Solving the Problems] In order to achieve the above object, the present invention is characterized by having a bubble removing means for removing bubbles present in the buffer solution in the electrophoresis tank.

[Operation] When a buffer solution is injected into the electrophoresis tank to form a parallel flow in the electrophoresis tank, and a DC voltage is applied to the electrodes in the electrolytic tank, and in this state, the electrophoresis sample is injected into the migration tank. The electrophoresis sample is decomposed into each component and separated.

At that time, if air bubbles are present in the buffer solution in the migration tank, the bubbles will rise to the liquid surface, but as mentioned above, since the buffer solution in the migration tank has a bubble removal means for removing the bubbles, , the air bubbles are removed by air bubble removal means.

Therefore, even if air bubbles are present in the buffer solution, the air bubbles can be reliably removed by the air bubble removal means.

Compared to conventional technology, which removes air bubbles after they have formed to a certain extent, it is possible to improve the separation performance of the electrophoresing sample, and also to increase the efficiency of separation work, since the removal can be done without shutting off the power.

[Example] An example of the present invention will be described below with reference to FIGS. 1 and 2. FIG. 1 is a front view of a longitudinal section showing an embodiment of the electrophoresis device of the present invention, and FIG. 2 is a side view of the same longitudinal section.

In the electrophoresis device of the embodiment, a migration tank 1 is formed in a main body 2, and electrolytic tanks A are formed on both sides of the migration tank 1.

To be more specific, the electrophoresis tank 1 is inside the main body 2 and has dimensions such that the height is larger than the width as shown in Fig. 1 in order to contain the electrophoresis sample and the buffer solution, and the depth is larger than the width as shown in Fig. 2. has smaller dimensions and defines a generally rectangular space.

The main body 2 is made of an insulating material such as acrylic, glass, or ceramics.

When the buffer solution is injected into the upper part of the electrophoresis tank 1 from the introduction hole 3 of the main body 2, it forms a parallel flow that flows straight downward from the upper part of the electrophoresis tank 1, and the electrophoresis sample flows in the same way due to the parallel flow. I have to. The introduction hole 3 communicates between the upper part of the electrophoresis tank 1 of the main body 2 and an external buffer tank (not shown).

On the other hand, the electrophoresis sample is a mixture of various components,
The buffer solution is injected into the migration tank 1 from the injection pipe 4 of the main body 2, and is caused to flow within the migration tank 1 along the parallel flow of the buffer solution. The injection pipe 4 is installed below the introduction hole 3 of the main body 2, and communicates between the migration tank 1 and an external migration sample tank (not shown).

Further, as shown in FIG. 2, a front tank wall 5a and a rear tank wall 5b are attached to the lower part of the migration tank 1 so as to face the front and rear parts of the migration tank 1. A preparative port 6 is provided at the bottom, a plurality of preparative tubes 7 are piped to the preparative port 6, and a discharge hole 8 for discharging the buffer solution is provided.

Further, on both the left and right sides of the electrophoresis tank 1, as shown in FIG. 1, anion exchange membranes 9. Electrolytic cells A and A are provided with a cation exchange membrane 10 in between. Each electrolytic cell A has anion exchange membranes 9 . It forms a space separated by the cation exchange membrane 10 and having approximately the same height as the front and rear tank walls 5a and 5b. In each electrolytic cell A, a live electrode 11 is provided on the anion exchange membrane 9 side, and an electrode 12 is provided on the cation exchange membrane 10 side, and the electrolyte is introduced into the introduction hole 13.
It enters the interior through the hole and flows out through the discharge hole 14. Live electrode 11 and negative electrode 12 are connected to corresponding poles of power supply 15.

Then, by turning on the power supply 17, both electrodes 11 and 12
When a DC voltage is applied to the migration tank 1, a buffer solution is injected into the migration tank 1 from the introduction hole 3 to form a vertical parallel flow in the migration tank 1, and in this state, the migration sample is introduced from the injection tube 4 into the migration tank 1.
When the electrophoresis sample is injected into the electrophoresis sample, each component of the electrophoresis sample moves and separates due to the difference between the moving force based on the amount of charge of each component of the electrophoresis sample and the fluid resistance that the particles of the electrophoresis sample receive,
Each separated component is fractionated into a fractionating tube 7 through a fractionating port 6.

Furthermore, a cooling tank 16 is provided so as to be able to circulate in order to cool down the Joule heat generated in the migration tank 1 when a voltage is applied to both electrodes 11 and 12. This cooling tank 16
As shown in FIG. 2, in the main body 2, the space is defined by facing the front tank wall 5a and the rear tank wall 5b, respectively.
Cooling water is supplied from the water introduction hole 17, while the water discharge hole 1
By being drained from the cooling tank 8, the water is circulated within the cooling tank 16. Therefore, the front tank wall 5a and the rear tank wall 5b are made of a material with high thermal conductivity in order to increase the cooling efficiency of the cooling tank 16.

In this electrophoresis apparatus, bubbles are generated in the buffer solution in the electrophoresis tank 1 when the buffer solution is injected into the electrophoresis tank 1 or for some other reason, and it is necessary to remove the bubbles.

Therefore, in the embodiment, a bubble removing means for removing bubbles in the buffer solution is provided.

The bubble removing means includes a gas passing filter 19 and a gas exhaust mechanism (not shown). Although not shown in detail, the gas passing filter 19 is a thin film in which a plurality of minute passages are formed, and is made of, for example, tetrafluoroethylene resin. The gas passing filter 19 is attached to an opening 2o formed in the upper part of the main body 2 with a filter holder 21, so that the gas passing filter 19 is disposed at a liquid level position in the migration tank 1. The filter holder 21 is inserted into the opening 2o of the main body 2.

The gas exhaust mechanism includes a vacuum pump 22 installed externally,
It consists of a suction pipe 23, one end of which is connected to the suction side of the vacuum pump 22, and the other end of which is connected to the opening 20 through a cap 24.

This air bubble removing means reduces the pressure inside the migration tank 1 by the suction force of the vacuum pump 22, and the air bubbles present in the buffer solution are sucked into the gas passing filter 19, and the air bubbles pass through the gas passing filter 19. And filter holder 21. By being sucked into the vacuum pump 22 through the suction pipe 23, the gas in the bubbles is discharged to the outside.

Therefore, the filter holder 21 is attached to the gas passing filter 19.
is formed to allow gas to pass through the microchannels of
For example, it is made of acrylic or the like.

Further, the inner wall 2a of the main body 2 near the liquid level in the migration tank 1 is formed in a chevron shape. That is, the chevron-shaped inner wall 2a forms a wide space at its lower part on the side of the introduction hole 3 for the buffer solution, and forms a space whose width gradually becomes narrower from the lower part to the top, and the top part forms a space for gas passage. It is located at a position corresponding to the lower center of the filter 19, so that bubbles of the buffer solution are guided to the gas filter 19 and collected therein.

Next, the operation of the electrophoresis apparatus of the example will be described.

A buffer solution is previously injected into the electrophoresis tank 1 through the introduction hole 3 to form a parallel flow within the electrophoresis tank 1, and the live electrode 11. - When a DC voltage is applied to the electrode 12 and the electrophoresis sample is injected into the electrophoresis tank 1 from the injection tube 4 in this state, the electrophoresis sample is decomposed into each component and passes from the aliquot port 6 to the separation tube 7. It will be fractionated.

At that time, if bubbles are present in the buffer solution in migration tank 1,
The bubbles float to the surface of the buffer solution. The floated air bubbles are removed by air bubble removal means. That is, the bubble removing means includes the gas passing filter 19 and the vacuum pump 22. Since it is equipped with a discharge mechanism consisting of a suction pipe 23, air bubbles in the buffer solution pass through the gas passing filter 19, and the filter presser 2
1. It is sucked into the vacuum pump 22 through the suction pipe 23 and discharged to the outside by the vacuum pump 22.

Therefore, even if there are air bubbles in the buffer solution, the air bubbles can be reliably removed by the air bubble removal means, compared to conventional techniques that remove air bubbles after they have formed to a certain extent.
Separation performance of electrophoretic samples can be improved. Moreover, since it can be removed without shutting off the power supply 15, the efficiency of separation work can also be improved.

In addition, since the inner part 112a near the liquid surface in the migration tank 1 has a mountain-shaped shape so as to guide air bubbles to the gas passing filter 19, the air bubbles are not scattered in the migration tank 1, and the air bubbles floating on the liquid surface are concentrated. This allows for more accurate removal.

[Effects of the Invention] As described above, according to claims 1 and 2 of the present invention, even if air bubbles exist in the buffer solution, the air bubbles can be reliably removed by the air bubble removal means. So,
The separation performance of the electrophoresis sample can be improved, and it can be removed without shutting off the power supply, and the efficiency of separation work can be improved. As a result, the separation and purification of the electrophoresis sample can be automated. According to claim 3, the inner wall near the liquid surface in the electrophoresis tank has a mountain-shaped shape and guides the bubbles floating on the liquid surface so that they can be easily removed by the bubble removing means, so that the bubbles can be removed accurately. effective.

[Brief explanation of drawings]

FIG. 1 is a front view of a longitudinal section showing an embodiment of the electrophoresis device of the present invention, and FIG. 2 is a side view of the same longitudinal section. DESCRIPTION OF SYMBOLS 1... Migration tank, 2... Main body, 2a... Inner wall of main body, 3... Buffer introduction hole, 4... Injection tube for electrophoretic sample, 6... Preparation port, 15 ... Power source, 19... Gas passing filter, 21... Filter holder, 22... Vacuum pump, 23 Suction pipe. Figure 2

Claims (1)

[Scope of Claims] 1. An electrophoresis apparatus characterized by having a bubble removing means for removing bubbles present in a buffer solution in a electrophoresis tank. 2. The bubble removing means includes a gas passing filter capable of passing air bubbles present in the buffer solution in the electrophoresis tank, and a discharge mechanism for discharging the bubbles to the outside through the gas passing filter. 1. The electrophoresis device according to 1. 3. The electrophoresis device according to claim 2, wherein the inner wall near the liquid surface of the electrophoresis tank has a chevron shape that guides air bubbles present in the buffer solution to a gas passing filter.