JP2001153841A - Electophoretic device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an electrophoretic device capable of performing a continuous separating and fractioning work of a sample by free flow electrophoresis and automatically separating and fractioning a plurality of subjects by a fractioning mechanism. SOLUTION: This device comprises a free flow electrophoretic element 10, a sample cup 49 arranged in the lower part of the outflow port of a separated liquid from the free flow electrophoretic element, and a mechanism for moving the position for retaining the sample cup in such a manner as to be approachable to and separable from the free flow electrophoretic element. According to this, an electrophoretic device capable performing a continuous separating and fractioning operation of a sample by free flow electrophoresis and automatically separating and fractioning a plurality of subjects by a fractioning mechanism can be provided. Further, taking one step ahead, a free flow electrophoretic device capable of continuously and easily separating and fractioning a sample from a minimum quantity at high speed and fully automatically separating and fractioning a plurality of samples can be provided.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to an electrophoresis apparatus, and more particularly, to a free-flow electrophoresis apparatus for separating a solute contained in a liquid sample by applying a voltage to the liquid sample.

[0002]

2. Description of the Related Art Electrophoresis can be used for separating and separating solutes (components) in a liquid sample and for analyzing a liquid sample. As an electrophoresis apparatus, an apparatus for performing separation using a gel or a membrane as a support or an apparatus for performing separation using a capillary is conventionally known.

For example, a capillary electrophoresis apparatus is
It can be composed of a capillary, a high voltage device, a detector and the like. The capillary is the heart of the electrophoresis apparatus, and generally, a fused silica glass elongated in a hollow cylindrical shape and reinforced by coating the periphery with a polyimide resin can be used.

[0004] Here, a comparison between the above-described apparatus using a gel or a membrane and the apparatus using a capillary shows that the latter capillary electrophoresis apparatus can be separated without using a support, and thus the sample after separation is used. It is not necessary to extract the separated substance from the support when fractionated and used. As the capillary electrophoresis device, a device already provided with a sorting mechanism is commercially available.

On the other hand, a free-flow electrophoresis method has been conventionally known as an electrophoresis method for performing separation and fractionation without using a support. In the free flow electrophoresis, a supporting liquid is caused to flow at a constant flow rate in a liquid flow path formed by sandwiching two plates to form a liquid flow to an outlet, and provided near a liquid inlet to the flow path. A sample is injected from the sample inlet, and a voltage is applied to the electrodes provided at both ends of the flow path.The substance contained in the sample flows out to the outlet while being electrophoretically separated on the positive or negative electrode side by the principle of electrophoresis. An electrophoresis apparatus utilizing this is a free-flow electrophoresis apparatus.

For example, JP-A-5-203623 (Document 1) discloses an electrophoresis tank having a plurality of outlets on the downstream side, a buffer solution tank having an outlet on the upstream side of the electrophoresis tank, and a buffer solution having the same. A low-pressure pressurizing device connected to the tank, a sorting container provided corresponding to each of the plurality of outlets, and a pipe having fluid resistance means for connecting the plurality of outlets and the sorting container respectively are provided. Free-flow electrophoresis devices have been reported.

[0007] The electrophoresis apparatus has the following features. In the above configuration, the buffer liquid tank is pressurized at a predetermined constant pressure by a low-pressure pressurizing device. Then
The buffer solution flows into the electrophoresis tank from the upstream outlet at a constant flow rate. On the other hand, the buffer solution, together with the sample separated from the plurality of outlets on the downstream side, flows into the collection container through the pipe. At this time, by the fluid resistance means of the pipe,
The flow velocity flowing through each pipe is kept constant. Therefore,
The flow velocity in the electrophoresis tank is made uniform. It is reported that by doing so, the flow can be made uniform with a simple device, and a compact and inexpensive electrophoresis device can be provided.

[0008]

However, the following points can be pointed out in the conventionally known electrophoresis apparatus based on the following considerations by the present inventors.

(A) First, consider a capillary electrophoresis apparatus as the electrophoresis apparatus described first. In such an apparatus, the technically most difficult point in the separation of separated components is a high voltage. There is a problem of operability accompanying the application. Further, in the capillary electrophoresis apparatus, the amount of components that can be separated by one analysis is extremely small. Therefore, in order to obtain a large amount, it is necessary to repeat the fractionation and accumulate the fraction, however, capillary electrophoresis has a low repetition accuracy, and this is also a problem. Becomes

(B) On the other hand, free flow electrophoresis, which allows continuous separation and separation of samples, has a problem in operability of a liquid flowing out of the free flow electrophoresis element.
For example, in the above cited document 1 (Japanese Patent Application Laid-Open No. 5-203623), the fractionated liquid is configured to flow into the fractionation vessels provided respectively corresponding to the plurality of outlets. No means for replacing the dispensing container as is clarified is specified.

Here, according to the recognition of the inventor, which is the basis of the idea and realization of the present invention, in order to establish an electrophoresis apparatus using a free-flow electrophoresis element as an analyzer, a plurality of electrophoresis apparatuses are required. It would be desirable to be able to separate the sample automatically (more preferably fully automated), but this would require replacement of the collection vessel. Further, when a plurality of samples can be analyzed, it is necessary to wash the free-flow electrophoresis element between the samples. However, if the analysis container is fixed, even the washing liquid is contained in the container. If the sorting mechanism is provided via a pipe corresponding to a plurality of outlets, the structure is complicated, and the sample container needs to be replaced for each sample and each washing, which is suitable for automation. Absent.

(C) Therefore, it is more desirable that continuous separation / separation work of a sample can be easily performed and a plurality of specimens can be automatically separated / separated. Is an electrophoresis device that can continuously and rapidly separate and separate samples from extremely small amounts, and can automatically separate and separate multiple samples, especially by free-flow electrophoresis. The device can be obtained.

(D) Furthermore, according to the recognition of the inventor, which is the basis of the idea and realization of the present invention, it is important for free-flow electrophoresis that the flow in the element is actually uniform. However, in Reference 1 mentioned above, there is a description referring to “uniform flow” (Reference 1, page 2, right column, lines 46 to 47, page 3, right column, third line, etc.). Even though it is described as being uniform, there is no function or confirmation means for confirming whether the flow is really uniform. For this reason, it cannot be determined whether or not a uniform flow actually occurs. Also,
In the free-flow electrophoresis, the separated solution is diluted with the supporting solution because of the characteristic of performing the electrophoretic separation while flowing the sample together with the supporting solution. Therefore, when the fractionated liquid is used for other purposes, it is desirable to concentrate the diluted liquid. However, the above-mentioned literature 1 does not particularly describe the concentration of such a fraction. Furthermore, in free-flow electrophoresis, since a voltage is applied to the electrophoresis tank having a larger area than in the above-described capillary electrophoresis, there is a risk that a large current flows. However, the above-mentioned document 1 does not specifically describe measures for avoiding the danger.

Accordingly, the present invention, based on the above considerations,
In addition, based on the considerations described later, this was made to improve the electrophoresis apparatus, and it is possible to continuously and easily separate and sort samples, and to automatically separate and sort multiple samples. It is an object of the present invention to provide an improved electrophoresis apparatus. Another object of the present invention is to provide an electrophoresis apparatus capable of confirming a flow in an element, particularly, an electrophoresis apparatus based on free-flow electrophoresis. Another object of the present invention is to provide an electrophoresis apparatus capable of concentrating a fraction, particularly a free-flow electrophoresis apparatus using free-flow electrophoresis. Another object of the present invention is to provide an electrophoresis apparatus having safety associated with voltage application, particularly, a free-flow electrophoresis apparatus using free-flow electrophoresis.

[0015]

According to the present invention, the following electrophoresis apparatus is provided. That is,
According to the present invention, the free-flow electrophoresis device has a free-flow electrophoresis device, and a sample cup is disposed below an outlet of a separation liquid from the free-flow electrophoresis device. An electrophoretic device is provided, which has a mechanism for moving the element so that the element can move forward and backward (claim 1).

Here, the “part holding the sample cup” in claim 1 corresponds to the separation and extraction mechanism of the sampling device in a preferred embodiment of the present invention (including modifications and variations of the first embodiment). According to the present invention, there is provided an electrophoresis apparatus capable of continuously separating and collecting samples by free-flow electrophoresis and capable of automatically separating and separating a plurality of samples by a sorting mechanism. be able to.

Further, according to the present invention, the free-flow electrophoresis element is disposed in an upper portion of the apparatus, and has an observation unit having a camera for observing a flow in the free-flow electrophoresis element. An electrophoresis device is provided (claim 2). Here, the "camera" in claim 2
In a preferred embodiment of the present invention (including modifications and variations of the second embodiment), a CCD camera is applicable.

According to the present invention, in addition to the functions and effects of the electrophoretic device of the first aspect, it is possible to provide a free-flow electrophoretic device capable of confirming the flow in the element. For example, by using a sample containing a fluorescent reagent as the sample, the flow of the liquid in the electrophoresis separation tank can be observed in real time as the flow of the fluorescent dye. Therefore, it is possible to proceed with the separation and separation while always observing the flow of the liquid, which is very important in the separation using the free-flow electrophoresis element. In this case,
In a preferred example, for example, a dichroic mirror is provided above the free-flow electrophoretic element, and a light source that causes light to enter the free-flow electrophoretic element via the dichroic mirror and a reflection from the free-flow electrophoretic element As a configuration having a camera that observes light through the dichroic mirror, the present invention can be suitably implemented, but without using a dichroic mirror or a light source, for example, as a mode of directly observing the flow of a dye as a color with a camera, The present invention can be suitably implemented.

Further, according to the present invention, there is provided an electrophoresis apparatus, wherein a portion for holding the sample cup is constituted by a heat block. According to the present invention, it is possible to provide a free-flow electrophoresis apparatus capable of concentrating a fractionated liquid, in addition to the operation and effect of the electrophoresis apparatus according to claim 1 or 2. In the present invention, the provision of the heat block makes it possible to control the temperature of the fraction, and if the heat block is used in a state where the temperature is raised to a certain temperature, the fraction can be concentrated. Therefore,
In the separation using the free-flow electrophoresis element, the fractionated solution diluted with the supporting solution can be concentrated simultaneously with the separation and fractionation operation.

Further, according to the present invention, there is provided an electrophoresis apparatus characterized in that the parts that come into contact with the liquid are made of non-metal (claim 4). According to the present invention,
In addition to the functions and effects of the electrophoretic device according to claim 1, 2, or 3, it is possible to provide a free-flow electrophoretic device having safety associated with voltage application. In the present invention, by configuring the portion where the liquid comes into contact with a nonmetal, it is possible to provide an electrophoresis apparatus having safety associated with voltage application. In this case, in a preferred embodiment, the present invention can be suitably implemented as a configuration in which a power supply for voltage application used for free flow electrophoresis is independent of other mechanisms.

[0021]

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

[Structure of the First Embodiment] FIGS. 1 to 4 show an embodiment of the electrophoresis apparatus of the present invention. FIG. 1 shows a configuration diagram of the electrophoresis apparatus. FIGS. 2 and 3 are diagrams for explaining a separation and extraction mechanism of the apparatus.
Indicates a free-flow electrophoresis element applicable to a separation and extraction mechanism. FIG. 1 also shows an overall configuration of an electrophoresis apparatus according to the present invention. In this embodiment, an electrophoresis apparatus 100 includes a sampling apparatus 102.
And a power supply for voltage application 103 and a computer for control 104.

As will be described later, the sampling device 102 and the voltage applying power source 103 are independent of each other, and are connected by a cord 110. The control computer 104 is also connected to the voltage application power supply 103 by the cord 110, and includes transmission of a control signal in order to cause the sampling device 102 including various control elements to automatically perform processing in a process described later. 1 is connected to a main body (main body housing) indicated by reference numeral 101 in FIG. 1 through connection means including a multi-core cable or the like (not shown) formed by a connector for signal transmission / reception. Things.

The sampling device 102 here has a sample stocker 1 as shown transparently therein.
05, the liquid sending pump 106, and the sampling mechanism 107
And a sampling liquid injection mechanism 108, and a separation / extraction mechanism 109 which is the heart of the apparatus as shown in FIGS. (Shown in a flat rectangular shape). This apparatus can be configured to be automatically controlled by the control computer 104.

Here, the sample stocker 105 can arrange a plurality of containers 105a as shown in the figure, and can set a sample. As described in detail below, the liquid sending pump 106 is connected to the control computer 1.
04 can be started at a predetermined timing,
The feeding of the supporting liquid can be performed by this. The sampling mechanism 107 can also be started by the control computer 104 at a predetermined timing. Sampling of a required amount of the sample in the sample stocker 105 and delivery to the sampling liquid injecting mechanism 108 can be performed by the sampling mechanism 107 thus activated, and the sampled liquid is transferred to the sampling liquid injecting mechanism 1.
The application to the separation and extraction mechanism 109 by 08 is also executed under the control of the control computer 104.

FIG. 2 is a side view of the separation / extraction mechanism 109, FIG. 3 is a top view of the separation / extraction mechanism 109, and FIG. 4 is an example of the applicable free-flow electrophoresis element 10. The free-flow electrophoresis element 10 incorporated in the separation and extraction mechanism 109 is shown.

As shown in FIG. 2, the free-flow electrophoresis element 10 includes, for example, two glass substrates 10a and 10a.
a, a gap serving as the electrophoresis separation tank 11 is formed between the two glass substrates 10a, 10a. Further, as shown in FIG. 3 or FIG. 4, a plurality of (seven in the illustrated example) support liquid inlets 12 are provided at one end of the gap (electrophoresis separation tank 11), and the opposite side is provided. At the end,
A plurality (six in the illustrated example) of separated liquid extraction ports 14 are provided. Similarly, the support liquid inlet 12 and the separated liquid outlet 14
Between the sample inlet 13 and the support liquid inlet 12 between
Are provided, and electrodes 15, 15 are provided on both side portions between the sample injection port 13 and the separated liquid extraction port 14.

In the illustrated example, a support liquid inlet 12 for injecting a support liquid, a sample inlet 13 for injecting a sample,
And the separated liquid extraction port 14 for fractionation are both on the same surface side, that is, the lower glass substrate 10a in FIG.
It is provided in. Here, FIG. 4 is a diagram showing the free-flow electrophoresis element 10 as a separation column incorporated in the separation / extraction mechanism 109 in FIG. 2 in a single state (as viewed from the lower side of the glass substrate portion). In this case, the electrophoresis separation tank 11 and the electrode 15 correspond to a state where the electrophoresis separation tank 11 and the electrode 15 are transmitted through the glass part). The portion of the free-flow electrophoresis element 10 shown in the right part of FIG. 3 has a glass on the upper surface so as to show a correlation with a movement position (a virtual line position) of a movable sorting mechanism 48 described later. The substrate portion is drawn with this removed.

The support liquid inlet 12 of the free-flow electrophoresis element 10 is connected to a liquid feed pump 106 in the main body housing by a liquid pipe 106a. When the liquid feed pump operates, the support liquid sent from the liquid feed pump 106 is supplied. The liquid is separated electrophoresis tank 11
Is injected into. The sample injection port 13 of the free-flow electrophoresis element 10 is connected to the sampling liquid injection mechanism 108 in the main body housing by a liquid tube 108a, whereby the sample is injected into the separation electrophoresis tank 11. The electrodes 15, 15 of the free-flow electrophoresis element 10 are connected to a voltage applying power supply so that a voltage can be applied from a voltage applying power supply 103 separate from the sampling device 102,
Thus, a voltage for free-flow electrophoresis is applied to the separation / electrophoresis tank 11.

In this embodiment, the upper surface of the sampling device 102 (the upper surface
The separation / extraction mechanism 109 configured in 1a) includes, in addition to the above-described free-flow electrophoresis element 10, a holding table 26 for the free-flow electrophoresis element 10, a waste vat 37 for effluent, It consists of the sorting mechanism 48 touched.

The free-flow electrophoretic element 10 is held on the holding table 26 with the support liquid inlet 12, the sample inlet 13, and the separated liquid extraction port 14 on the lower side. Is done. Here, it is preferable to adopt a configuration in which the holding table 16 fixes the periphery of the free-flow electrophoresis element 10. By doing so, the upper surface of the separation and migration tank 11 (upper glass substrate 10a)
The free-flow electrophoresis element 10 can be configured so as to be able to hold the free-flow electrophoresis element 10 in a state where there is no support from the lower surface (the lower glass substrate 10a) (a state where a space can be left on the upper and lower sides). Therefore, as will be described further below, the liquid can be injected and fractionated from the lower surface of the separation and migration tank 11, and the separation and migration tank 11 can be observed from the upper surface.

Here, the former advantage (the point that the liquid can be injected and separated from the lower surface of the separation / electrophoresis tank 11) is that the separation mechanism 48 is used to flow the separated liquid from the free-flow electrophoresis element 10. 2 and 3 to the lower side of the separated liquid extraction port 14 serving as an outlet.
From the state of the solid line position (48a) to the virtual line position (48b).
When moving by sliding as shown in
At this time, it is advantageous to make the movement be performed without any obstacles that may hinder such movement. Further, the latter advantage (the point that the separation / electrophoresis tank 11 can be observed from the upper surface) is exhibited and utilized as a further advantage in another embodiment described later.

The waste vat 17 is provided below the separation liquid extraction port 14 and receives the effluent from the separation liquid extraction port 14 and flows to a waste liquid tank (not shown) via the liquid pipe 37a. Have.

The sorting mechanism 48 has a sample cup and is configured to be able to move forward and backward with respect to the free-flow electrophoretic element 10 (see arrows S in FIGS. 2 and 3). Here, it has a sample cup 49 for preparative sample and a sample cup holder 50, and the sample cup 49 for preparative process is stored in the sample cup holder 50,
The sample cup holder 50 has a notch extending in a space 51 provided in the sorting mechanism 48 (here, as shown in FIG. 3, extending in a direction orthogonal to the sorting mechanism moving direction S (vertical direction in FIG. 3). (Elongated hole). Preferably, the mounting position (the position along the longitudinal direction of the long hole) in the long hole can be adjusted.

The sorting mechanism 48 is configured to be driven in parallel with the free-flow electrophoresis element 10 by, for example, a cylinder (such as an air cylinder) having a retractable rod (not shown). As described above, it is configured to move horizontally with respect to the free-flow electrophoresis element 10 horizontally positioned on the main body 101. Therefore, the sorting mechanism 48 can slide forward and backward on an appropriate base on the upper surface of the apparatus. Here, the driving range of the sorting mechanism 48 is at least
From the position where the tip of the sorting mechanism 48 driven toward the end is separated from the end of the holding table 26 to the position where the space 51 is located below the separated liquid extraction port 14, and the driving source thereof is not shown as described above. This is done by using a cylinder. The driving range is a position (48a) indicated by a solid line to a position (48b) indicated by a virtual line in FIGS.

Sample cup holder 5 of sorting mechanism 48
0 can be provided by the number of the separation liquid extraction ports 14 (six in the illustrated example) (FIG. 3 shows an example in which the number of the separation liquid extraction ports 14 is provided). 49
Is installed at a position where the effluent from the separated liquid extraction port 14 is dropped into each sample cup when the sorting mechanism 48 is at the position (48b) indicated by the phantom line and fits.

The sampling device 102 is constructed so as to be capable of automatically processing not only separation and separation of samples but also cleaning and surface treatment of the free-flow electrophoretic element 10 between samples. Here, when these processes are also performed automatically, control for the processes can also be executed under the control of the control computer 104.

The control computer 104 is control means for controlling the entire electrophoresis apparatus 100 (system), and includes an input / output interface, an operation processing circuit (CPU)
And various control programs executed by the arithmetic processing circuit;
And a storage circuit for storing and storing various calculation results, and is provided with input means such as a keyboard and a display unit 104a.

The sampling device 102 and the voltage applying power source 103 are independent devices.
Is connected to earth. In this case, all parts of the sampling device 102 that are in contact with the liquid are made of non-metal, and the electrodes 1 of the free-flow electrophoresis element 10
The voltage from 5 does not leak to the equipment making up the device.

Next, a method for separating and collecting a sample using the electrophoresis apparatus 100 will be described. It is assumed that a program for each condition is set in the control computer 104 in advance, and the control program is stored in the storage circuit of the control computer 104.

A sample is set in the sample stocker 105, and a control program for executing the following steps is started. In the sample stocker 105,
A container 105a containing a sample for a plurality of specimens can be set in advance.

At the start, the sorting mechanism 48 is held at a position (48a) shown by a solid line (first position on the left side in FIGS. 2 and 3: initial position). The step of attaching / detaching (automatic exchange) of the sample cup 49 for fractionation to the sample cup holder 50, which will be described later, can be executed when it is at such a position. The state of the sorting mechanism 48 in the solid line state (48a) shown in the left part of the figure is already in the sample cup holder 5
A state in which a new sample cup 49 for preparative separation has been attached to 0 has been shown.

Here, under the control of the control computer 104, the liquid feed pump 106 is started according to the control program, and the support liquid inlet 1 of the free-flow electrophoretic element 10 is opened.
The support liquid is sent from the second to the electrophoresis separation tank 11 to create a liquid flow up to the separation liquid extraction port 14.

At this time, the effluent from the separated liquid extraction port 14 is dropped onto the waste liquid vat 37, and as a result, is led to a waste liquid tank (not shown). At this point, the sorting mechanism 48 is still in operation.
Therefore, the preparative sample cup 49 on the sample cup holder 50 has not moved, and therefore, the preparative sample cup 49 does not exist below the separated liquid extraction port 14. As a result, the supporting liquid at this time is a waste liquid vat 37.
The liquid is not dropped unnecessarily into the sample cup 49 for fractionation. It is at the timing described later that the liquid to be dispensed to the dispensing sample cup 49 is dispensed.

Next, under the control of the control computer 104, the sampling mechanism 107 is activated to sample a specified amount of the sample contained in the container 105a in the sample stocker 105. The sampled liquid is sent to the sampling liquid injection mechanism 108, and a specified amount of the sample is injected into the sample injection port 1 of the free-flow electrophoresis element 10.
3 will be injected.

After the sample is injected, a voltage is applied from the voltage applying power source 3 to the electrodes 15 and 15 of the free-flow electrophoretic element 10 under the control of the control computer 104, and a voltage is applied between the electrodes 15 and 15. Then, the sample flows out to the separation liquid extraction port 14 while performing electrophoresis toward the positive or negative electrode side depending on its electric characteristics. In accordance with this, under the control of the control computer 104,
When the separation of the sample occurs, the cylinder (not shown) of the sorting mechanism 48 is driven, and therefore, the position (4
The sorting mechanism 48 is moved to 8b) (the second position in a state where it is movable to the right in FIGS. 2 and 3: the sorting position). As a result, at such a timing, the liquid separated by voltage application is:
The liquid is dropped from the separated liquid extraction port 14 to the sample cup 49 for fractionation. The total amount of the injected sample is injected,
At the time when the liquid flows out to the position 4, the sorting mechanism 48 moves to the position (4
Return to the position of 8a). As mentioned above, at the return position of the sorting mechanism 48, the operation of automatically changing the sample cup for sorting can be performed. Such return control is also executed under the control of the control computer 104.

Here, in the present electrophoresis apparatus 100, the sample cup 49 for sorting can take the first and second positions with the movement of the sorting mechanism 48, instead of the configuration as described in the above-mentioned document 1. Then, one of the positions (first position) replaces the old and new sample cups 49, and the other position (second position) sorts the sample cups 49. It becomes possible. Even in the case of continuous separation / separation of samples, it can be appropriately performed and easily performed, and also in the case of separation / separation of multiple samples,
This can be automated by the sorting mechanism 48, and the separation and sorting can be easily performed automatically as described below.

When the return control is performed, the preparative sample cup 49 containing the separated and extracted liquid is moved by a robot arm mechanism (not shown) and guided to another process or stored for each application. You. Thus, the preparative sample cup containing the separated and extracted liquid for exchanging the preparative sample cup is removed. In this case, the command control for the robot arm mechanism is also executed under the control of the control computer 10.

Further, in this embodiment, after the sample is injected,
In the same procedure, the cleaning liquid and the surface treatment liquid flow in the electrophoresis separation tank 11 via the sampling mechanism 107 and the sampling liquid injection mechanism 108, and the cleaning and the surface treatment are performed automatically. At this time, the effluent from the separated liquid extraction port 14 is
To the waste liquid tank. Also at this time, the liquid is passed through as it is. Of course, also in this case, in the present electrophoresis apparatus 100, the sample cup for separation does not exist below the separation liquid extraction port 14, and
As a result, the washing liquid and the like are not stored in the sample cup for fractionation.

Here, in the above-described processing such as washing, if the container is configured to be fixed, the washing liquid may be contained in the container, or may be connected to a plurality of outlets via piping. In such a case, the configuration is complicated, and it is necessary to replace the collection container for each sample and each washing, which is not suitable for automation. However, the electrophoresis apparatus 100 does not have such disadvantages. .

Thus, for example, during the above-mentioned cleaning or surface treatment, the above-mentioned sample cup (49) for sorting is returned to the sorting mechanism 48 returned to the solid line position (48a).
Can be removed, and subsequently, a process of mounting a new sample cup 49 for sample collection to the sample cup holder 50 can be executed by the control of the robot arm mechanism. (Attaching a new preparative sample cup for exchanging the preparative sample cup) Thus, the automatic exchange of the preparative sample cup is also easily performed. After such replacement, the state described first is obtained. Therefore, when the continuous processing is subsequently performed, the series of procedures described above can be automatically performed in the same manner.

Thus, all of the above steps are performed automatically by a control program built in the control computer 104, and a continuous processing of a plurality of samples can be performed by repeating this procedure.

[Operation / Effect of First Embodiment] The electrophoresis apparatus 100 according to the present embodiment, as described in the above configuration, is capable of continuous separation / separation of samples by free-flow electrophoresis. It is possible to provide an electrophoresis apparatus in which a plurality of specimens can be easily separated and fractionated by the fractionation mechanism 48 easily. Therefore also
A good and effective solution can also be realized in view of the considerations of known electrophoresis devices at the beginning of the description.

As described above, in the capillary electrophoresis apparatus, the most difficult technical point in separating the separated components is the operability associated with the application of a high voltage.
Because the amount of components that can be separated in a single analysis is extremely small,
In order to obtain a coherent amount, it is necessary to repeat the fractionation and accumulate the fraction, but capillary electrophoresis does not have high repetition accuracy, which is a problem.
In the known free-flow electrophoresis device, the operability of the liquid flowing out of the free-flow electrophoresis element becomes a problem,
According to the above configuration of the apparatus 100 of the present embodiment, such a problem can be solved and the electrophoresis apparatus is effective.

In particular, in order to establish an electrophoresis apparatus using a free-flow electrophoresis element as an analyzer, it is desirable to be able to automatically separate a plurality of samples. For this purpose, it is necessary to replace a preparative container. Where necessary, in the case of the above-mentioned document 1, the fractionated liquid is configured to flow into containers (fractionating containers) provided respectively corresponding to the plurality of outlets, and therefore, there is no need to replace the container. If you do not have it, and if you want to be able to analyze multiple samples, you need to wash the free-flow electrophoresis element between samples, but if the container is fixed, the washing liquid will be stored in the container Furthermore, the sorting mechanisms provided via pipes corresponding to the plurality of outlets have a complicated structure, and the sample container is replaced for each sample and each washing. Not suitable for automation be necessary.

On the other hand, in configuring an electrophoresis apparatus for electrically separating and separating a liquid sample, a sampling mechanism 107 for a plurality of samples, a sampling liquid injection mechanism 108,
It has a power supply 103 for voltage application, a liquid feed pump 106, and a computer 104 for control (FIG. 1), and has a free-flow electrophoresis element 10 as a separation column, and the free-flow electrophoresis element 10 is arranged in an upper part of the apparatus. An outlet for the separated liquid from the free-flow electrophoresis element 10 is formed on the lower substrate 10a of the free-flow electrophoresis element 10, and a sample cup 49 is arranged below the outlet to hold the sample cup 49. It has a mechanism that allows the part to move back and forth with respect to the free-flow electrophoresis element 10 (Figs. 2 to 4), and enables continuous, high-speed, and easy separation and fractionation of samples from a very small amount. Thus, the apparatus 100 of the present embodiment, which can provide a free-flow electrophoresis apparatus capable of automatically separating and separating a plurality of samples, can be provided at the beginning of the specification (a). ~ It is also advantageous problem solution from the viewpoint mentioned in (c), so it is possible to realize an improved electrophoresis apparatus advanced one step.

In addition, the apparatus 100 according to the present embodiment can provide an electrophoresis apparatus having safety associated with voltage application. Comparing capillary electrophoresis and free-flow electrophoresis in terms of these aspects, the latter free-flow electrophoresis has the danger that a large current will flow because a voltage is applied to the electrophoresis tank, which has a larger area than capillary electrophoresis. Although there is no particular description of the measures for avoiding the danger in the above-mentioned document 1, the electrophoresis which makes the safety accompanying the voltage application by configuring all the parts in contact with the liquid with a non-metal is described. Example device 10 capable of providing the device
0 can be realized as having both of the effective measures for avoiding the danger described in the consideration (d) at the beginning of the specification, and therefore, in this regard, the improved electrophoresis which has taken a step further The device can be realized.
In this case, if the power supply 103 for voltage application is made independent of the sampling device 102, it is an even more effective workaround to secure the safety associated with voltage application and avoid danger.

[Modifications and Modifications of the First Embodiment] The constitutions of this embodiment can of course be variously modified and changed.

[1-1]: For example, in the present embodiment, the free-flow electrophoresis element 10 has two glass substrates 10
The material is not limited to glass but may be any insulating substrate such as plastic.

[1-2]: Further, for example, the support liquid inlet 12, the sample inlet 13, and the separated liquid extraction port 14 are formed on the same surface of the free-flow electrophoresis element 10, but the formed surface is Not limited. For example, the separated liquid extraction port 14 serving as an outlet may be formed on a side surface of the free-flow electrophoresis element.

[1-3]: Also, for example, the sorting mechanism 4
8 is configured to be driven by a cylinder, but the drive source is not limited to the cylinder.

[1-4]: Also, for example, the sample cup 49 for sample collection and the sample cup holder 50 are used to separate the required sample solution from the sample solution flowing out of the separation solution extraction port 14 so that the sample solution flows out. It is only necessary to provide the same number of liquid extraction ports, and it is not always necessary to provide the same number of separation liquid extraction ports 14. In this case, the outflow portion of the fractionated liquid that does not require fractionation passes through the space (elongated hole) 51 in the fractionation mechanism 48 as it is (that is, the fractionation sample cup 4).
9 and the portion where the sample cup holder 50 does not exist) are dropped onto the waste liquid vat 37 and similarly guided to the waste liquid tank (see FIG. 3).

In the slit-shaped space (elongated hole) 51 provided in the sorting mechanism 48, the sample cup holders 50 are thus mounted so that the number of the sample cup holders or the position on the sorting mechanism 48 can be adjusted. In combination with the structure that can be configured as
It has not only such attachment means but also such a function.

The points [1-1] to [1-4] also apply to the following modifications and changes in the embodiment.

[Structure of the Second Embodiment] Another embodiment (second embodiment) of the electrophoresis apparatus of the present invention will be described below with reference to FIG.
This will be described with reference to FIG. Electrophoresis apparatus 1 according to first embodiment
Reference numeral 00 denotes an electrophoresis apparatus that electrically separates and separates a liquid sample in which a plurality of solutes are dissolved, and includes at least a sampling mechanism for a plurality of samples, a sampling liquid injection mechanism, a power supply for voltage application, a liquid sending pump, and a control computer. Having, also has a free-flow electrophoresis element as a separation column, the free-flow electrophoresis element is horizontally arranged at the top of the apparatus,
An outlet for the separated liquid from the free-flow electrophoresis element is formed on the lower surface of the free-flow electrophoresis element, a sample cup is disposed below the outlet, and a portion for fixing the sample cup is provided on the free-flow electrophoresis element. While this was an example of having a mechanism to move horizontally with respect to the electrophoretic element,

The following is an example in which the flow in the free-flow electrophoretic element can be further confirmed in addition to this, and is considered as a modification of the first embodiment. Can also.

FIG. 5 is for explaining the main part of the preferred embodiment in this case. The electrophoresis apparatus according to the present embodiment (second embodiment) also has the basic structure of the first embodiment. As in the example, the sampling device 102 shown in FIGS.
And a power supply for voltage application 103 and a computer for control 104. Here, FIG.
Is a diagram of a sampling device 102A in the present embodiment. This is a view corresponding to FIG. 2 of the first embodiment. The difference from the first embodiment is that the observation mechanism denoted by reference numeral 120 in FIG. This is an addition to the configuration, and the other components described so far other than the observation mechanism 120 may be the same. Accordingly, the same components are denoted by the same reference numerals, and detailed description thereof will be omitted. In the description of the first embodiment, for example, “sampling device 102” is replaced with “sampling device 102A” in the description. This is applied to the description of the present embodiment (this point
The following examples are based on this.) Hereinafter, the main part of the electrophoresis apparatus 100 according to the present embodiment will be described.

As shown in FIG. 5, here, a system including an excitation light source 121, a dichroic mirror 122, an absorption filter 123, and a CCD camera 124 is provided above the sorting mechanism 109 of the first embodiment shown in FIG. Is configured. The dichroic mirror 121, as described above,
Free flow electrophoresis element 1 with no support on its upper surface
At the top of 0. The excitation light source 121 is connected to the free-flow electrophoresis element 10 via the dichroic mirror 122.
The CCD camera 124 is a camera for observing the reflected light from the free-flow electrophoretic element 10 via the dichroic mirror 122, and the absorption filter 123 is a
It is arranged between the camera and the CD camera 124. Here, CCD
The image information from the camera 124 may be provided to the control computer 104 (FIG. 1), for example.

The observation mechanism 120 in the electrophoresis apparatus 100 having the above configuration can be used as follows.

For example, when a sample containing a fluorescent dye is used as the sample, the light from the excitation light source 121 is applied to the free-flow electrophoretic element 10 via the dichroic mirror 122, whereby the dye contained in the sample is Is excited. The fluorescence is dichroic mirror 1
The light is reflected by the CCD camera 12 and cut by the absorption filter 123 to a wavelength other than the fluorescence wavelength of the sample.
4 observed. That is, when a sample containing a fluorescent reagent is used as a sample in the present apparatus, the flow of the liquid in the electrophoresis separation tank 11 can be observed in real time as the flow of the fluorescent dye.

In the case where the image information captured by the CCD camera 124 is provided to the control computer 104 shown in FIG. 1, the information may be displayed on the display unit 104a, or may be used instead of or together with this. This may be displayed on an appropriate monitor display device.

[Operation and Effect of Second Embodiment] The electrophoresis apparatus 100 according to the present embodiment has the above-mentioned configuration of the first embodiment,
The same operations and effects as those of the first embodiment described in [Operations and effects of the first embodiment] are obtained.

Further, in addition to this, for example, by using a sample containing a fluorescent reagent as the sample, the electrophoresis separation tank 1 can be used.
1 can be observed in real time as a flow of a fluorescent dye. This makes it possible to proceed with the separation and fractionation operation while always observing the flow of the liquid, which is very important in the separation using the free-flow electrophoresis element 10. As described above, it is also mentioned in the above-mentioned reference 1 that it is important that the flow in the element is uniform in the free-flow electrophoresis, but there is no means for confirming whether the flow is really uniform. For this reason, the present embodiment which can provide a free-flow electrophoresis apparatus capable of confirming the flow in the element 10 can be provided in contrast to those not adopting the present invention, which cannot determine whether a uniform flow is generated. The device 100
The present invention can also be realized as having appropriate confirmation / judgment means from the viewpoint described in the consideration (d) at the beginning of the specification.

[Modifications and Modifications of the Second Embodiment]
Naturally, the configuration of the embodiment can also be variously modified and changed (including [1-1] to [1-4] in the above-mentioned section [Modifications and Modifications of First Embodiment]).

[2-1]: For example, the CCD camera 12
4 is not particularly limited as long as it is a camera capable of observing a fluorescent image.

[2-2]: For example, the positions of the excitation light source 121 and the CCD camera 124 are not particularly limited.
It can be arranged at any position where the fluorescent image of the free-flow electrophoresis element 10 can be observed.

[2-3]: For example, in the illustrated example, the dichroic mirror 122 is provided above the free-flow electrophoresis element 10.
And a CCD camera 124 for observing the reflected light from the free-flow electrophoretic element 10 through a dichroic mirror 122. Not limited. For example, instead of using the excitation light source 121, the dichroic mirror 122, the absorption filter 123, and the like, it is also possible to directly observe the flow of the dye as a color with the CCD camera 24. The present invention can also be implemented in this way.

[Structure of Third Embodiment] Next, still another embodiment (third embodiment) of the electrophoresis apparatus of the present invention will be described.
This will be described with reference to FIG. The present embodiment is an example in which, when the sample is collected in the sample cup for separation, it is intended to be able to concentrate the collected liquid, and a modification of the first embodiment. You can think of it as an example.

FIG. 6 is a view for explaining the main part of the preferred embodiment in this case. The electrophoresis apparatus according to the present embodiment (third embodiment) also has the basic structure of the first embodiment. As in the example, the sampling device 102 shown in FIGS.
And a power supply for voltage application 103 and a computer for control 104. Here, FIG.
Is a diagram of the sampling device 102B in the present embodiment, which corresponds to FIG. 2 of the first embodiment. Hereinafter, the main part of the electrophoresis apparatus 100 according to the present embodiment will be described.

As shown in FIG. 6, here, instead of the sample cup holder 50 provided as means for holding the sample cup 49 in the sorting mechanism 109 of the first embodiment shown in FIG. It is configured by providing a block 61. As in the case of the sample cup holder 50 in FIG. 2, the heat block 61 has a hole for holding the sample cup 49, and can be implemented as a configuration in which a plurality of heat blocks are provided for each sample cup.

Here, the heat block 61 can control the temperature for each block. According to the present embodiment, when the heat block 61 is introduced, the temperature control can also be performed by the control of the control computer 104. In this case, the temperature control is performed as a control program. Can be incorporated in advance.

The heat block 61 in the electrophoresis apparatus 100 having the above configuration can be used as follows. The control computer 104 controls the temperature of the heat block 61. Under the control of the control computer 104, the sample cup 49 is held while the heat block 61 is heated to a certain temperature. When the separation and preparative work described in the section of the embodiment is performed, the liquid flowing out of the separation liquid extraction port 14 of the free-flow electrophoresis element 10 is subjected to the sample cup 49 heated by the heat block 61.
Is dropped. Therefore, the fractionated liquid dropped into the sample cup 49 is heated by the heat of the heat block 61, the volatilization of the fractionated liquid is promoted, and the fractionated liquid is concentrated.
In other words, this configuration enables the fractionated liquid to be concentrated simultaneously with the separation and fractionation operation.

[Operation / Effect of Third Embodiment] The electrophoresis apparatus 100 according to the present embodiment has the above-mentioned configuration of the first embodiment,
The same operations and effects as those of the first embodiment described in the section "Operation and effects of the first embodiment" can be obtained.

Further, in addition to this, heat block 6
By providing 1, temperature control of the fractionated liquid can be performed. If the heat block 61 is used in a state where the temperature is raised to a certain temperature, the fractionated liquid can be concentrated. This allows the fractionated solution diluted with the support solution in the separation using the free-flow electrophoresis element 10 to be concentrated simultaneously with the separation and fractionation operation. As described above, in the case of free flow electrophoresis, the separated liquid is diluted with the support liquid due to the characteristic of performing electrophoretic separation while flowing the sample together with the support liquid, and therefore, when the separated liquid is used for other purposes. Although it is desirable to concentrate the diluted liquid, for example, there is no description about the concentration of the preparative liquid in the above-mentioned literature 1, but it is intended to provide a free-flow electrophoresis apparatus capable of concentrating the preparative liquid. The possible embodiment device 100 can be realized as having effective means from the viewpoint described in the consideration (d) at the beginning of the specification.

[Modifications and Modifications of the Third Embodiment]
Naturally, the configuration of the embodiment can be variously modified and changed (including [1-1] to [1-4] in the section of [Modifications and Modifications of First Embodiment]).

[3-1]: For example, heat block 6
1 is not particularly limited as long as it can control the temperature.

[3-2]: For example, the temperature can be changed to a thermostat using a liquid or the like.

[3-3]: In the above description, the heat block 61 is heated and used. However, if it is desired to cool the fractionated liquid, the temperature can be lowered and cooled.

[3-4]: In this embodiment, it is needless to say that the main part can be implemented in the form shown in FIG.
This can be combined with the second embodiment (including [2-1] to [2-3] in the section of [Modifications and Modifications of the Second Embodiment]), and needless to say. Absent. In such a combination, the same operation and effect as those of the second embodiment described in the section of [Operation and effect of the second embodiment] are further obtained. The present invention can also be implemented in this way.

The contents described above can be considered as the following inventions.

[Appendix 1] An electrophoresis apparatus which electrically separates and separates a liquid sample in which a plurality of solutes is dissolved, includes a sampling mechanism for at least a plurality of specimens, a sampling liquid injection mechanism, a power supply for voltage application, A pump, a control computer, and a free-flow electrophoresis element as a separation column, wherein the free-flow electrophoresis element is horizontally disposed at the top of the apparatus, and the separation from the free-flow electrophoresis element is performed. A liquid outlet is formed on a lower surface or a side surface of the free flow electrophoretic element, a sample cup is arranged below the outlet, and a portion for fixing the sample cup is horizontally positioned with respect to the free flow electrophoretic element. An electrophoresis apparatus or system having a moving mechanism.

[Appendix 2] In the electrophoresis apparatus or system according to Appendix 1, a dichroic mirror is provided above the free-flow electrophoresis element, and light is applied to the free-flow electrophoresis element via the dichroic mirror. An electrophoresis apparatus or system comprising: an incident light source; and a camera for observing reflected light from the free-flow electrophoresis element through the dichroic mirror.

[Appendix 3] The electrophoresis apparatus or system according to Appendix 1 or 2, wherein the portion for fixing the sample cup is constituted by a heat block. .

[Additional Item 4] In the electrophoresis apparatus or system according to any one of the additional items 1 to 3, the power supply for voltage application is independent of other mechanisms, and the liquid comes into contact with the liquid supply device. An electrophoresis apparatus or system wherein all parts are non-metallic.

[Brief description of the drawings]

FIG. 1 is a diagram showing an electrophoresis apparatus according to one embodiment (first embodiment) of the present invention.

FIG. 2 is a diagram provided for describing an example of a separation and extraction mechanism in the same example.

FIG. 3 is a diagram also used for the explanation.

FIG. 4 is a view showing an example of a free-flow electrophoresis element that can be incorporated in the same mechanism.

FIG. 5 is a view showing an electrophoresis apparatus according to another embodiment (second embodiment) of the present invention, and is used for describing a main part in the same example.

FIG. 6 is a view showing an electrophoresis apparatus according to still another embodiment (third embodiment) of the present invention, and is used for describing a main part in the same example.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 10 Free flow electrophoresis element 10a Glass substrate 11 Electrophoresis separation tank 12 Support liquid injection port 13 Separation liquid extraction port 14 Sample injection port 15 Electrode 26 Holder 37 Waste liquid vat 37a Liquid tube 48 Preparative mechanism 49 Sample cup 50 Sample cup holder 51 Space (elongated hole) 61 Heat block (sample cup holder) 100 Electrophoresis apparatus 101 Main body 102, 102A, 102B Sampling apparatus 103 Power supply for voltage application 104 Control computer 104a Display section 105 Sample stocker 105a Container 106 Liquid supply pump 106a Liquid Tube 107 Sampling mechanism 108 Sampling liquid injection mechanism 108a Liquid tube 109 Separation and extraction mechanism 110 Code 120 Observation mechanism 121 Light source for excitation 122 Dichroic mirror 123 Absorption filter 124 CCD camera

Claims (5)

[Claims] 1. A free-flow electrophoresis device, comprising a free-flow electrophoresis device, a sample cup disposed below an outlet of a separation solution from the free-flow electrophoresis device, and a portion for holding the sample cup. An electrophoresis apparatus, comprising a mechanism for moving forward and backward with respect to the electrophoresis. 2. The electrophoresis device according to claim 1, wherein the free-flow electrophoresis element is disposed in an upper portion of the apparatus, and further includes an observation unit including a camera for observing a flow in the free-flow electrophoresis element. apparatus. 3. The electrophoresis apparatus according to claim 1, wherein a portion for holding the sample cup is constituted by a heat block. 4. The electrophoresis apparatus according to claim 1, wherein a part to which the liquid comes in contact is made of a nonmetal. 5. The electrophoresis apparatus according to claim 4, wherein a power supply for applying voltage for free flow electrophoresis is independent of other mechanisms.