JP5047928B2 - Electrophoresis device and components thereof - Google Patents

Description

  The present invention relates to an electrophoresis apparatus and its constituent instruments, and more specifically, to an electrophoresis apparatus and its constituent instruments for concentrating and separating a substance to be separated in a gel.

  After the completion of the Human Genome Project, proteome research has been actively conducted. The proteome is intended for a specific cell, organ, or the entire protein produced by translation in an organ, and proteome research includes protein profiling and the like.

  The proteome is a mixture of many proteins, each protein having unique properties in surface charge and molecular weight. For this reason, it is possible to separate more proteins with high resolution by separating them in combination rather than separating proteins depending only on charge or molecular weight alone. As such a technique, two-dimensional protein migration is most often used.

  Two-dimensional electrophoresis is electrophoresis that separates a sample in the presence and absence of a denaturing agent, and is an excellent technique that can separate several hundreds of proteins at a time (for example, Patent Document 1 and Non-Patent Document 1). Patent Document 1). Two-dimensional electrophoresis consists of two types of electrophoresis: isoelectric focusing that separates proteins depending on electric charge, and slab gel electrophoresis (SDS-PAGE) that separates proteins depending on molecular weight.

  SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) is a general electrophoresis method using SDS, which is a kind of anionic surfactant, as a protein solubilizer. There are many types of SDS-PAGE, but the Laemmli method is most frequently used at present (see, for example, Non-Patent Document 2). In the Laemmli method, a gel for concentrating proteins (concentration gel) is used in addition to a gel for separation (separation gel).

  A conventional gel used in SDS-PAGE will be described below with reference to FIG. FIG. 8 is a perspective view showing the gel 301 for concentration and the gel 302 for separation.

As shown in FIG. 8, in the conventional gel, a concentration gel 301 is arranged between a sample introduction region 303 and a separation gel 302. When electrophoresis is performed using such a gel, the electrode on the concentration gel 301 side is set to a negative (−) potential, and the electrode on the separation gel 302 side is set to a positive (+) potential. At this time, the sample equilibrated by SDS moves from the minus direction toward the plus direction, but the mobility of the sample in the separation gel 302 is smaller than the mobility of the sample in the concentration gel 301. The sample is concentrated at the interface between the concentration gel 301 and the separation gel 302. For this reason, the sample separated in the separation gel 302 forms a clear separation band.
JP 11-30605 A (published on February 2, 1999) Edited by Amersham Pharmacia Biotech, “The latest protocol for electrophoresis for more sensitive and quantitative detection analysis, from general operation to gelomics and proteomics”, Yodosha, 2000, p. 55-108 Nature, (USA), 1970, Vol. 227, p. 680-685

  However, making a concentrating gel on a separating gel complicates the structure of the entire gel, so it is difficult to produce the gel with high reliability.

  The present invention has been made in view of the above-described problems, and an object thereof is an electrophoresis apparatus capable of concentrating a sample and obtaining a clear separation band without using a concentration gel in protein electrophoresis. Is to provide.

  As a result of intensive studies, the present inventors have found that a substance to be separated can be concentrated by using a concentration electrode that applies a voltage to the substance to be separated introduction region of the gel without using a concentrated gel, thereby completing the invention. It was.

  That is, the electrophoresis apparatus according to the present invention is an electrophoresis apparatus that concentrates a substance to be separated introduced into a gel at a specific position of the gel and then separates the separated substance in the separation direction of the gel. The first concentration electrode is provided at a position where the substance to be separated is concentrated at the specific position.

  According to the said structure, the said to-be-separated substance can be electrophoresed to the said specific position. As a result, the substance to be separated is collected at the specific position and concentrated at the position.

  In order to perform electrophoresis of the substance to be separated to the specific position, the substance to be separated is sandwiched between the first concentrating electrode and another electrode in the vicinity of the specific position, and a voltage is applied between the electrodes. May be applied. For example, if the substance to be separated is a protein equilibrated with SDS, it has a negative charge, so by applying a voltage in the direction from the first concentration electrode to the other electrode, The substance to be separated can be electrophoresed toward the first concentration electrode.

  As the other electrode, for example, another concentration electrode provided on the opposite side to the first concentration electrode with respect to the substance to be separated may be used, or an electrophoresis apparatus is originally provided. A separation electrode provided may be used. That is, since the electrophoresis apparatus for separating the substance to be separated in the gel includes a pair of separation electrodes so as to sandwich the gel, at least one of the separation electrodes and the first concentration electrode together with the above-described substance to be separated. A separation substance is sandwiched, and the separation electrode can be used as the other electrode.

  Then, if normal electrophoresis is performed after the substance to be separated is concentrated as described above, the substance to be separated concentrated at the specific position is separated to form a clear separation band. Therefore, if the electrophoresis apparatus according to the present invention is used, the analysis accuracy of the substance to be separated can be improved without preparing a special gel.

  In addition, the electrophoresis apparatus according to the present invention preferably further includes a second concentration electrode that is paired with the first concentration electrode and sandwiches the substance to be separated introduction region into which the substance to be separated is introduced. .

  According to the above configuration, by applying a voltage between the first and second concentration electrodes, the substance to be separated introduced into the substance introduction region can be successfully concentrated at the specific position. In the above configuration, since the separation electrode provided in the electrophoresis apparatus does not have to be used as an electrode paired with the first concentration electrode, switching of electrical connection is not necessary.

  In the electrophoresis apparatus according to the present invention, the first and second concentration electrodes have a linear shape, and the first and second concentration electrodes are arranged in parallel and orthogonal to the separation direction. It is preferable.

  According to the above configuration, the first and second concentration electrodes are linear electrodes that are arranged in parallel to each other perpendicular to the separation direction, and thus have an influence on the voltage during separation in the electrophoresis apparatus. There is almost no. For this reason, a to-be-separated substance can be isolate | separated suitably irrespective of presence of the 1st and 2nd concentration electrode.

  Further, the concentrated substance to be separated becomes a line orthogonal to the separation direction, and a suitable separation pattern can be obtained.

  The electrophoresis apparatus according to the present invention preferably further includes a substrate that supports the gel, and the first and second concentration electrodes are preferably formed on a support surface of the gel on the substrate. Moreover, it is preferable that an active element for switching the first and second concentration electrodes is formed on the support surface of the substrate.

  According to the above configuration, the first and second concentration electrodes can be formed by semiconductor technology. In addition, the voltage applied between the first and second concentration electrodes can be quickly turned on and off by the switching function of the active element.

  Furthermore, in the electrophoresis apparatus according to the present invention, it is preferable that the active element is composed of continuous grain boundary crystalline silicon, low-temperature polysilicon, high-temperature polysilicon, or amorphous silicon.

  According to the above configuration, the active element can be easily formed as a part of the circuit.

  Further, in the electrophoresis apparatus according to the present invention, a transfer film is superimposed on the gel, and further includes a pair of transfer electrodes sandwiching the gel and the transfer film, and the transfer electrode is separated from the separation electrode. It is preferable to include a plurality of electrode regions arranged along the direction, a rectifying element that connects the plurality of electrode regions, and a switching mechanism that switches between positive and negative of the bias applied to the rectifying element.

  In the above configuration, after the substance to be separated in the gel is concentrated and separated, when a positive bias is applied to the rectifying element, a current flows through the rectifying element, and each electrode region is electrically connected. . At this time, a voltage is applied between the pair of transfer electrodes. At this time, if a voltage is applied using one of the pair of transfer electrodes as the anode on the transfer film side, the separated substance to be separated moves to the transfer film and is transferred to the transfer film.

  During the concentration and separation of the substances to be separated in the gel, if a negative bias is applied to the rectifier element, no current flows through the rectifier element, and each electrode region is electrically insulated. The Therefore, the presence of the transfer electrode does not adversely affect the concentration and separation of the substance to be separated.

  According to the above configuration, concentration, separation, and transfer of a substance to be separated can be easily performed in one apparatus.

  Moreover, the component instrument of the electrophoresis apparatus according to the present invention is a component instrument included in the electrophoresis apparatus according to the present invention, and includes the substrate, the first and second concentration electrodes, and the active element. It is characterized by that.

  According to the said structure, the component instrument which concerns on this invention can be provided in an electrophoresis apparatus as a detachable component. The electrophoresis apparatus provided with the component instrument according to the present invention can achieve the same effects as the electrophoresis apparatus according to the present invention.

  The electrophoresis method according to the present invention includes an introduction step of introducing a substance to be separated into a substance separation region of a gel, a first concentration electrode disposed at a specific position on the gel, and the first concentration. A concentration step of concentrating the substance to be separated by applying a voltage in a specific direction in the substance introduction region to be separated by a second concentration electrode sandwiching the substance introduction region to be separated with the electrode; and the concentration Subsequent to the step, the method includes a separation step of applying a voltage in the specific direction in the gel to separate the substance to be separated in the specific direction in the gel.

  In the introduction step, the substance to be separated is introduced into the substance to be separated introduction region of the gel. The substance to be separated is negatively charged in the gel.

  In the concentration step, the first concentration electrode arranged at a specific position on the gel and the second concentration electrode paired with the first concentration electrode and sandwiching the separated substance introduction region, A voltage is applied in a specific direction in the substance introduction region. At this time, if a voltage is applied using the first concentration electrode as an anode, the substance to be separated moves to the first concentration electrode, but cannot travel beyond the first concentration electrode. For this reason, the substance to be separated is concentrated in the vicinity of the specific position.

  In the separation step, following the concentration step, a voltage is applied in the specific direction in the gel to separate the substance to be separated in the specific direction in the gel. The separated substance to be separated forms a clear separation band.

  Therefore, if the electrophoresis method according to the present invention is used, the analysis accuracy of the substance to be separated can be improved.

  According to the electrophoresis apparatus according to the present invention, since the first concentration electrode for concentrating the substance to be separated in the gel is provided, a clear separation result can be obtained without using the concentration gel. Can improve the reproducibility of electrophoresis results.

(Two-dimensional electrophoresis apparatus 100)
An embodiment of the present invention will be described below with reference to FIGS. In the present embodiment, the electrophoretic device 10 is described as being incorporated in the two-dimensional electrophoretic device 100, but the present invention is not limited to this and may be used alone.

  First, a schematic configuration of the two-dimensional electrophoresis apparatus 100 will be described with reference to FIG. FIG. 2 is a schematic diagram showing the two-dimensional electrophoresis apparatus 100 according to the present embodiment.

  As shown in FIG. 2, a two-dimensional electrophoresis apparatus 100 includes a first-dimensional electrophoresis region 101 that separates a sample by an isoelectric point, and a second-dimensional electrophoresis region 102 (electrophoresis that separates a substance to be separated by molecular weight. The apparatus 10) and an immune reaction region 103 for detecting a sample with high specificity of antigen-antibody reaction.

  Note that the sample (substance to be separated) in the present embodiment is a protein extract, but the present invention is not limited to this and may be any substance to be analyzed by electrophoresis. Specifically, a preparation from a biological material (for example, an individual organism, a body fluid, a cell line, a tissue culture, or a tissue fragment) can be preferably used, and a polypeptide or a polynucleotide can be more preferably used. it can.

  In the first-dimensional electrophoresis region 101, a sample introduction and swelling tank 106, an isoelectric focusing tank 107, and an SDS equilibration tank 108 are provided. In the sample introduction and swelling tank 106, the sample is filled into an immobilized pH gradient (IPG) gel 105 supported at the tip of the support 104.

  Isoelectric focusing is performed in the isoelectric focusing tank 107, and the sample in the immobilized pH gradient gel 105 is separated (Isoelectric Focusing, IEF) depending on the isoelectric point of the protein. In the SDS equilibration tank 108, SDS is introduced into the sample in the separated immobilized pH gradient gel 105.

  After the treatment in the SDS equilibration tank 108 is completed, the immobilized pH gradient gel 105 is carried to the electrophoresis apparatus 1 and placed in the sample introduction region 7 (see FIG. 1) of the gel (SDS-PAGE gel) 6. The In the electrophoresis apparatus 10, SDS-PAGE electrophoresis is performed. As will be described later, transfer may be performed in the electrophoresis apparatus 10.

  After the processing in the electrophoresis apparatus 10 is finished, the gel 6 or the transfer film transferred from the gel 6 is transferred to the reaction layer 109 of the immune reaction region 103, and suitable sample analysis (antigen-antibody reaction or the like) is performed.

(Electrophoresis device 10)
Next, the configuration of the electrophoresis apparatus 10 will be described below.

  As shown in FIG. 2, the electrophoresis apparatus 10 includes buffer solution tanks 2 and 3, separation electrodes 4 and 5, and a substrate 8. Buffer tanks 2 and 3 are used to fill the buffer. As the buffer solution, a buffer solution having a composition generally used for electrophoresis may be used. A separation electrode 4 is provided in the buffer solution tank 2, and a separation electrode 5 is provided in the buffer solution tank 3. On the region sandwiched between the buffer baths 2 and 3, a pair of substrates 8 are disposed so as to sandwich the plane of the gel 6. In addition, this invention is not restricted to this, One board | substrate 8 may be arrange | positioned so that the gel 6 may be supported.

  The gel 6 is preferably a polyacrylamide gel suitable for protein separation, but the present invention is not limited to this and may be a gel generally used for electrophoresis, such as an agarose gel. When the separation electrode 4 is a cathode and the separation electrode 5 is an anode, the gel 6 is arranged so that the sample introduction region 7 is located on the separation electrode 4 side.

  Next, the board | substrate 8 is demonstrated below with reference to Fig.1 (a) (b). FIG. 1A is a perspective view showing the substrate 8 and the gel 6, and FIG. 1B is a top view thereof.

  As shown in FIGS. 1A and 1B, a concentration electrode (second concentration electrode) 11 and a concentration electrode (first concentration electrode) 12 are formed on the substrate 8. The concentration electrode 12 is provided in the vicinity of a specific position where the substance to be separated in the gel 6 is to be concentrated. Here, the “position where the substance to be separated is concentrated at a specific position” where the concentration electrode is arranged in this specification means that the substance to be separated is concentrated at a predetermined position by applying a voltage to the concentration electrode. The concentrating electrode is a position of the concentrating electrode, and a person skilled in the art appropriately adjusts the concentrating electrode from the position of the concentrating electrode and the position of the substance to be separated to the position where the substance to be separated is concentrated at a specific position Can be arranged. The concentration electrodes 11 and 12 are provided on the plane of the substrate 8 on the side in contact with the gel 6 so as to sandwich the sample introduction region 7. Moreover, the shape of the concentration electrodes 11 and 12 is linear, and is arrange | positioned in parallel orthogonally to the separation direction of a sample. At this time, it is preferable that the substrate 8 on which the concentration electrodes 11 and 12 are formed is configured to be detachable as the component instrument 1 of the electrophoresis apparatus 10.

  The concentration electrodes 11 and 12 are electrically connected to the power source 13. The material of the concentration electrodes 11 and 12 may be a material having conductivity, and is preferably a material that does not deteriorate even when brought into contact with a buffer solution. Specifically, although not limited to these, platinum, gold, copper, zinc, or the like can be used. The power supply 13 may be built in the substrate 8 using a semiconductor manufacturing technique or a thin display device manufacturing technique.

  The operation of the electrophoresis apparatus 1 is as follows. Although the description will be made assuming that the concentration electrode 11 is a cathode and the concentration electrode 12 is an anode, these may be reversed.

  First, after a sample is introduced into the sample introduction region 7 of the gel 6, the power supply 13 is turned on and a voltage is applied between the concentration electrodes 11 and 12. At this time, a voltage is applied for a certain time until the sample is concentrated at the specific position near the concentration electrode 12 (anode). After the sample is concentrated near the concentration electrode 12, it is preferable to turn off the power supply 13 and cut off the voltage between the concentration electrodes 11 and 12. Next, the power source connected to the separation electrodes 4 and 5 is turned on, and a voltage is applied between the separation electrodes 4 and 5. This initiates sample separation.

  By the above operation, the sample in the gel 6 is concentrated and separated, and a clear separation band can be obtained.

  Other circuits or the like of the concentration electrodes 11 and 12 may be formed on the substrate 8.

  For example, as shown in FIG. 5, an active element 17 that switches the concentration electrodes 11 and 12 is preferably formed on the substrate 8. At this time, the concentration electrode 12 is connected to the power source 13 via the active element 17. FIG. 5A is a perspective view showing a component instrument 30 composed of the substrate 8 on which the active element 17 is formed, and FIG. 5B is a circuit diagram thereof.

  As shown in FIG. 5B, the active element 17 has a triode structure of an anode, a cathode, and a gate electrode, the gate electrode is connected to the concentration electrode 12, and the anode (or cathode) is an X electrode. 18 and the cathode (or anode) is connected to the Y electrode 19.

  The process of providing the active element 17 on the concentration electrode 12 will be described below with reference to FIG. 6A to 6D are process sectional views of the semiconductor layer forming process.

  First, a thin film serving as a material for the gate electrode / wiring 202 is formed on the substrate 8. A glass substrate is preferably used as the substrate 8, but a plastic substrate such as an acrylic plate or a film such as PET (Polyethylene Terephthalate) may be used. As the gate electrode / wiring material 202, tantalum (Ta), molybdenum (Mo), aluminum (Al), or an alloy thereof can be used. For forming a thin film of the material of the gate electrode / wiring 202, it is preferable to use a plasma CVD method (Chemical Vapor Deposition) or a sputtering method, but a PE-CVD method (Plasma Enhanced Chemical Vapor Deposition) is used. May be. After forming a thin film of the material of the gate electrode / wiring 202, it is patterned into a desired pattern by photolithography and etching.

Next, as shown in FIG. 6A, a gate insulating film 203 is formed using a SiNX (silicon nitride) film or the like. Further, a semiconductor layer 204 is formed over the gate insulating film 203. As the semiconductor layer 204, a-silicon (amorphous (amorphous) state silicon), CG (Continuous Grain, continuous grain boundary crystal) silicon, low-temperature or high-temperature poly (polycrystalline) silicon, and impurity-doped silicon thereof. Can be used.
The gate electrode / wiring 202, the gate insulating film 203, and the semiconductor layer 204 are preferably formed continuously in vacuum from the viewpoint of cleanliness at the interface between the layers.

  Next, as shown in FIG. 6B, a source electrode / wiring 205 and a drain electrode / wiring 206 are formed. As materials for the source electrode / wiring 205 and the drain electrode / wiring 206, it is preferable to use a low-resistance material in order to reduce the influence of signal delay. For example, an Al (aluminum) based alloy can be used. In the contact portion between the source electrode 205 and the drain electrode 206 of the semiconductor layer 204, it is preferable to reduce contact resistance with the source electrode 205 and the drain electrode 206 by adding a high concentration impurity to the semiconductor layer. The active element 17 is formed by the above process.

Next, as shown in FIG. 6C, an interlayer insulating film (protective film) 207 is formed on the active element 17. The interlayer insulating film 207 plays a role of protecting the active element 17, and a SiNX film can be used. However, when the active element 17 is required to be flat, it is preferable to use SiO ₂ (silicon oxide). At this time, the SiO ₂ can be planarized by polishing by CMP (Chemical Mechanical Polishing) or by reflowing by BPSG (Boronic Phosphoric Silicate Glass).

  Next, as shown in FIG. 6D, the through hole 208 and the concentration electrodes 11 and 12 are formed. A through hole 208 is provided on the drain electrode / wiring 206 to electrically connect the drain electrode 206 and the concentration electrode 12.

  The active element 17 formed as described above functions as a TFT (Thin Film Transistor). The operation when the active element 17 is an n-type TFT will be described below.

  When the active element 17 is in the ON state, a positive bias is applied to the gate electrode 202, and the semiconductor layer 204 is in the n + state. At this time, a state between the source electrode 205 and the drain electrode 206 is in the (n +) − (n +) − (n +) state, and a current flows.

  On the other hand, when the active element 17 is in the OFF state, a negative bias is applied to the gate electrode 202, the semiconductor layer 204 becomes p-type, and the distance from the source electrode to the drain electrode is the (n +)-(p)-(n +) state. Thus, no current flows.

  Therefore, the concentration electrodes 11 and 12 can be quickly switched by switching the active element 17 between the ON state and the OFF state.

  In other embodiments, the concentration electrode 11 may not be provided. An equivalent effect can be obtained by using the separation electrode 4 instead of the concentration electrode 11. That is, if a voltage is applied between the separation electrode 4 and the concentration electrode 12, the substance to be separated can be electrophoresed at the specific position.

  In addition, the place where the concentration electrodes 11 and 12 are provided is not limited to that shown in FIG. 1, and can be designed to be considered as the most suitable place by those skilled in the art if semiconductor manufacturing technology or thin display device manufacturing technology is used. . For example, in one embodiment, as shown in FIG. 3, it may be formed on the gel 6 side with respect to the sample introduction region 7. Fig.3 (a) is a perspective view which shows the other form of the component instrument 1, FIG.3 (b) is the top view. Thus, embodiments in which the concentration electrodes 11 and 12 are provided so as to sandwich a part of the region into which the substance to be separated is introduced are also within the scope of the present invention. Even in such a form, the substance to be separated eventually moves to a position sandwiched between the concentration electrodes 11 and 12 due to diffusion or the like, and is concentrated at the specific position.

  Furthermore, in another embodiment, as shown in FIG. 4, a stripe electrode 14 for transfer may be formed on the substrate 8. FIG. 4 is a perspective view showing a component instrument 20 composed of the substrate 8 on which the stripe electrode 14 is formed.

  The stripe electrode 14 has a structure in which a plurality of linear conductors are arranged in parallel. The material of this conductor may be a material having conductivity, and is preferably a material that does not deteriorate even when brought into contact with a buffer solution. Specifically, although not limited to these, platinum, gold, copper, zinc, or the like can be used. A diode (rectifier element) 15 that switches the stripe electrode 14 is provided between the conductors of the stripe electrode 14. The stripe electrode 14 and the diode 15 can be formed on the substrate 8 by using conventional semiconductor manufacturing technology or thin display device manufacturing technology.

  When electrophoresis and transfer are performed using the component instrument 20 shown in FIG. 4, the pair of substrates 8 are arranged so as to sandwich the gel 6 and the transfer film. Of the stripe electrodes 14 formed on the pair of substrates 8, the stripe electrode 14 formed on the substrate 8 on the transfer film side is used as an anode, and the stripe electrode 14 formed on the other substrate 8 is used as a cathode. .

  The transfer film may be made of a substance that can fix the sample, and a thin film is preferable. Specifically, a nitrocellulose membrane, a PVDF membrane, a nylon membrane, or the like used for a biopolymer analysis technique such as a well-known Western blotting method can be suitably used.

  The operation of performing electrophoresis and transfer using the component tool 20 is as follows. First, a negative bias is applied to the diode 15 to turn off the stripe electrode 14 and the sample is introduced into the sample introduction region 7 of the gel 6. Apply voltage. After the sample is concentrated, the voltage between the concentration electrodes 11 and 12 is turned off. Next, a voltage is applied between the separation electrodes 4 and 5 to separate the sample. After the sample is separated, the voltage of the separation electrodes 4 and 5 is turned off. Next, a positive bias is applied to the diode 15 to turn on the stripe electrode 14 and apply a voltage between the pair of stripe electrodes 14. As a result, the sample moves to the stripe electrode 14 side of the anode and is transferred from the gel to the transfer film.

  As described above, by using the component instrument 20 shown in FIG. 4, both electrophoresis and transfer can be performed in the electrophoresis apparatus 10.

  In one embodiment, as shown in FIG. 7, both the transfer stripe electrode 14 and the active element 17 described above may be formed on the substrate 8. FIG. 7 is a top view of a component instrument 40 including the substrate 8 on which the stripe electrode 14 and the active element 17 are formed. This component instrument 40 has the function by the stripe electrode 14 and the active element 17, and can perform each switching quickly.

  In one embodiment, a logic circuit, a memory circuit, or the like that controls energization to the active element 17 and the stripe electrode 14 may be further formed on the substrate 8. If this is formed on the substrate 8, it is possible to provide an electrophoretic device 10 with higher functionality.

(Electrophoresis method)
The electrophoresis method according to the present invention includes an introduction step of introducing a substance to be separated into a substance separation region of a gel, a first concentration electrode disposed at a specific position of the gel, and the first concentration electrode. A concentration step of concentrating the substance to be separated to the specific position by applying a voltage in a specific direction in the substance introduction area to be separated by a second concentration electrode sandwiched between the substance introduction area to be separated and Then, following the concentration step, a separation step of applying a voltage in the specific direction in the gel to separate the substance to be separated in the specific direction in the gel may be included. In the separation step, it is preferable not to apply a voltage between the concentration electrodes.

  The electrophoresis apparatus according to the present invention can be suitably used for an electrophoresis method in which a sample is concentrated and then separated.

(A) is a perspective view which shows the component instrument which concerns on one Embodiment of this invention, (b) is the top view. 1 is a side view showing a two-dimensional electrophoresis apparatus according to an embodiment of the present invention. (A) is a perspective view which shows the component instrument which concerns on one Embodiment of this invention, (b) is the top view. It is a top view which shows the component instrument which concerns on one Embodiment of this invention. (A) is a perspective view which shows the component instrument which concerns on one Embodiment of this invention, (b) is the circuit diagram. (A)-(d) is a figure which shows process sectional drawing of a semiconductor layer formation process. It is a top view which shows the component instrument which concerns on one Embodiment of this invention. It is a perspective view which shows the conventional gel for SDS-PAGE.

Explanation of symbols

1, 20, 30, 40 Component instrument 4, 5 Separation electrode 8 Substrate 10 Electrophoresis device 11, 12 Concentration electrode 14 Stripe electrode 15 Diode 17 Active element

Claims (4)

An electrophoretic apparatus for concentrating a substance to be separated introduced into a gel at a specific position on the gel and then separating the substance in the separation direction of the gel,
A first concentration electrode;
The first concentration electrode is provided at a position where the substance to be separated is concentrated at the specific position ,
A second concentrating electrode that is paired with the first concentrating electrode and sandwiches a substance introduction region into which the substance to be separated is introduced;
The shapes of the first and second concentration electrodes are linear, and the first and second concentration electrodes are arranged in parallel and perpendicular to the separation direction,
It further comprises a substrate that supports the gel,
The first and second concentration electrodes are formed on the support surface of the gel in the substrate,
An active element formed on the support surface and connected to the first or second concentration electrode;
The active element performs switching as to whether or not a voltage is applied between the first and second concentration electrodes,
The gel is overlaid with a transfer film,
It further includes a pair of transfer electrodes sandwiching the gel and the transfer film,
The transfer electrode is
A plurality of electrode regions arranged along the separation direction;
A rectifying element for connecting the plurality of electrode regions, and
An electrophoretic device comprising a switching mechanism that switches between positive and negative biases applied to the rectifying element . 2. The electrophoretic device according to claim 1 , wherein the active element is composed of continuous grain boundary crystalline silicon, low-temperature polysilicon, high-temperature polysilicon, or amorphous silicon. A component instrument of an electrophoresis apparatus provided in the electrophoresis apparatus according to claim 1 or 2 ,
A component of an electrophoresis apparatus, comprising the substrate, the first and second concentration electrodes , the active element , and the transfer electrode . An electrophoresis method using the electrophoresis apparatus according to claim 1 or 2,
An introduction step of introducing the object to be separated material to the be separated substance introduction region of the gel,
A concentration step of concentrating the substance to be separated by applying a voltage in the separation direction by the first concentration electrode and the second concentration electrode;
A separation step of applying a voltage in the separation direction in the gel and separating the substance to be separated in the separation direction in the gel following the concentration step.

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