JP2004279168A - Electrophoretic device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a chip-sized electrophoretic device capable of concurrently analyzing a lot of samples with high resolutions for a short period. <P>SOLUTION: The electrophoretic device 10 is provided with a substrate 20 which has a first main surface 20a and a second main surface 20b opposite to the first main surface, an electrophoretic channel 30 which is formed on the first main surface of the substrate 20, and an electrophoretic channel protecting film 40 which is formed on the first main surface having the electrophoretic channel and made up so as to expose both ends of the electrophoretic channel. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to an electrophoresis apparatus for separating and analyzing biological substances such as proteins and polynucleotides, and more particularly to an electrophoresis apparatus which is easy to handle and capable of separating and analyzing samples at high resolution.
[0002]
[Prior art]
With the development of microfabrication technology, for example, a technology for integrating a chemical analysis system into a chip made of a so-called chip size glass substrate of several centimeters square to analyze biomolecules has been developed.
[0003]
First, a device called a DNA chip or a DNA microarray was put to practical use. Such a device is intended to analyze a larger number of samples in a shorter time and with higher sensitivity by reducing the scale of execution as compared with a conventional chemical analysis system.
[0004]
In the future, it is foreseen that proteins will be the main analysis target. Conventionally, in protein analysis, protein molecules charged by an electric field are electrophoresed in a large-sized porous gel such as a polyacrylamide gel, and the proteins are separated by charges to perform analysis.
[0005]
In such a conventional analysis system, it is difficult to maintain the uniformity of the composition of the polyacrylamide gel, and there is a problem in reproducibility of analysis results.
[0006]
In order to solve such problems of gel composition uniformity and reproducibility of analysis results, a porous silicon layer is formed on a silicon substrate, a sealing material is disposed thereon, and a sample is placed on the porous silicon layer. There is known an electrophoresis apparatus which performs electrophoresis by infiltrating and applying an electric field to analyze proteins. (For example, refer to Patent Document 1 and Claims.)
[0007]
[Patent Document 1]
JP-A-9-304340
[0008]
[Problems to be solved by the invention]
In the analysis using the electrophoresis apparatus disclosed in the above-mentioned literature, it is difficult to analyze many samples simultaneously. In addition, since the migration length cannot be sufficiently obtained, it is difficult to increase the resolution and perform highly accurate analysis.
[0009]
Therefore, an object of the present invention is to provide a so-called chip-size electrophoresis apparatus that can analyze a large number of samples at the same time in a short time and with higher resolution while being separated from each other. .
[0010]
[Means for Solving the Problems]
In order to achieve this object, the electrophoresis apparatus of the present invention has the following structural features. That is, a first main surface, a substrate having a second main surface facing the first main surface, a plurality of grooves formed in the first main surface, and formed to fill the grooves. Channel formed of a porous material, first and second edges formed at both ends of the migration channel, and formed such that the first and second edges are exposed. And an electrophoresis channel protective film.
[0011]
According to the configuration of the electrophoresis apparatus of the present invention, electrophoresis can be performed on a scale of a chip size, and in a state where a plurality of migration channels are separated from each other. It can be analyzed with higher resolution.
[0012]
BEST MODE FOR CARRYING OUT THE INVENTION
Hereinafter, embodiments of the present invention will be described with reference to the drawings. In the drawings, the shapes, sizes, and arrangements of the components are only schematically shown to the extent that the present invention can be understood, and the present invention is not particularly limited thereby. Further, in the following description, specific materials, conditions, numerical conditions, and the like may be used, but these are only one of preferred examples and, therefore, are not limited thereto. In addition, it is to be understood that the same constituent components are denoted by the same reference numerals in the drawings used in the following description, and the overlapping description may be omitted.
[0013]
(First Embodiment)
With reference to FIGS. 1A, 1B and 1C, the configuration of the electrophoresis apparatus according to the first embodiment of the present invention will be described. FIG. 1A is a schematic plan view of the configuration of the electrophoresis apparatus according to the first embodiment as viewed from above, and FIG. 1B is a cross-sectional view taken along line II of FIG. FIG. 1C is a schematic cross-sectional view illustrating a cut surface cut along a broken line, and FIG. 1C is a schematic cross-sectional view illustrating a cut surface cut along a II-II ′ broken line in FIG. It is sectional drawing.
[0014]
The electrophoresis apparatus 10 according to the first embodiment includes a substrate 20. The substrate 20 has a first main surface 20a and a second main surface 20b facing the first main surface 20a.
[0015]
As the substrate 20, for example, a film-shaped body or a plate-shaped body conventionally applied to semiconductor devices such as a silicon substrate, a glass substrate such as quartz glass or non-fluorescent glass, or a plastic substrate formed of polyimide or the like can be appropriately selected. it can.
[0016]
Preferably, a silicon substrate is used. If a silicon substrate is used, for example, an electric circuit for applying a voltage for electrophoresis, a control circuit for controlling each of the electrophoresis channels 30 such as start and end of electrophoresis, a temperature and a substrate By forming a sensor element for detecting the inclination, and furthermore, an arithmetic circuit and a communication circuit by using a semiconductor manufacturing process, it is possible to achieve higher functionality.
[0017]
Here, the substrate 20 constituting the electrophoresis apparatus 10 of the present invention is, for example, a plate-like body in which the first main surface 20a and the second main surface 20b have the same shape and the same rectangular shape. Preferably, for example, the size of the first and second main surfaces 20a and 20b is set such that the length in the short direction is about 10 mm and the length in the long direction is about 30 mm.
[0018]
A plurality of grooves 22 are formed on the first main surface 20 a of the substrate 20. The groove 22 has a rectangular cross section in this configuration example, but is not limited to this, and may be formed in, for example, a semicircular shape or any other suitable shape. These grooves 22 are formed in a linear shape and in parallel with each other and apart from each other along the set longitudinal direction of the first main surface 20a.
[0019]
The size of the groove 22 can be appropriately set in consideration of the resolution and the sample to be applied within a range that does not impair the object of the present invention. As for the total length, the longer the migration distance is, the higher the resolution is, so it is preferable to make the total length as long as possible.
[0020]
At this time, the total length is preferably equal to the longitudinal length of the first main surface 20a. For example, when the length of the substrate 20 in the longitudinal direction is 30 mm as described above, the total length of the groove 22 is preferably 30 mm.
[0021]
The size of the cross section of the groove 22, that is, the groove width on the first main surface 20 a may be appropriately set in consideration of the amount of the sample to be electrophoresed, for example, the amount of the protein or nucleic acid that is a biomolecule to be injected or the measurement method. . For example, when a protein is assumed as a sample, its width is preferably set in the range of 0.5 μm to 1000 μm. Preferably, this width is, for example, about 10 μm.
[0022]
Similarly, the depth (height) of the groove 22 is preferably set in the range of 0.1 μm to 1000 μm when a protein is assumed as a sample. This depth is preferably set to, for example, about 3 μm.
[0023]
The distance between the adjacent migration channels 30 may be determined in consideration of the number of migration channels to be formed within a range that does not impair the object of the present invention. Can be used, so that this interval can be, for example, in the range of 0.1 μm to 10 μm. Alternatively, any suitable interval larger than 10 μm may be used. Preferably, this interval is set to, for example, about 100 μm in consideration of ease of sample injection and analysis.
[0024]
An electrophoresis channel 30 is formed in each of the plurality of grooves 22. The migration channel 30 is formed by embedding the groove 22 with a porous material. This porous body is preferably made of, for example, porous silica.
[0025]
These migration channels 30 have a linear shape along the longitudinal direction of the first main surface 20a, and are formed so as to be parallel to each other and apart from each other. That is, the first migration channel 30-1, the second migration channel 30-2, the third migration channel 30-3,..., And the n-th migration channel 30-n (where n is 4 or more) are linear. Are formed so as to be parallel to each other and apart from each other.
[0026]
On the first main surface 20a of the substrate 20 on which the migration channel 30 is formed, both end portions 32 of the migration channel 30, that is, first and second edge portions 32a and 32b are exposed. That is, the migration channel protective film 40 is formed such that the first and second edge portions 32a and 32b are exposed.
[0027]
According to the electrophoresis device of the first embodiment, electrophoresis can be performed in a state of being separated for each of the plurality of migration channels, so that a large number of samples can be simultaneously separated in a short time. And at a higher resolution. In particular, since the electrophoresis channels are formed independently so as to be spaced apart from each other, for example, a plurality of unrelated samples can be electrically transferred without adverse effects such as simultaneous interference of the plurality of electrophoresis channels and mixing of the samples. Electrophoresis can be performed.
[0028]
Next, a method for manufacturing the electrophoresis apparatus according to the first embodiment will be described with reference to FIGS.
[0029]
FIGS. 2A to 3B are schematic cross-sectional views each showing a part of a cut surface taken along the line II-II ′ of FIG.
[0030]
In describing the method for manufacturing the electrophoresis apparatus 10 of the present invention, only one electrophoresis apparatus 10 is illustrated and described in each of the drawings. However, in actuality, for example, a large number of A plurality of electrophoresis apparatuses 10 are manufactured on a large-sized substrate 20 at the same time as in the case where the semiconductor devices are formed in a lattice shape.
[0031]
First, regions for forming a plurality of electrophoresis devices 10 are set in advance on a large-sized substrate 20 (not shown). In this region, the spacing between the semiconductor devices 10, the shape and width of the migration channels 30, and the spacing between adjacent migration channels 30, that is, the number of migration channels 30 are set, assuming that the migration will be performed later. I do.
[0032]
Next, as shown in FIG. 2A, the semiconductor device is applied in a conventionally known semiconductor device manufacturing process so that a region where an electrophoresis channel is formed is exposed on the first main surface 20a of the substrate 20. The first main surface 20a of the substrate 20 is covered by a photolithography process under the same conditions as described above, using a resist pattern obtained by patterning the resist layer 70 (hereinafter, may be simply referred to as a resist layer) as a mask. As a material for the resist layer 70, a silicon-containing positive resist is preferably used. Then, the resist layer 70 is formed by performing a spin coat application, a pre-bake treatment, a photomask formation, an exposure and a development process.
[0033]
Specifically, first, a resist material is applied to the entire surface of the substrate by spin coating to form a film, and a pre-bake process is performed as necessary. Next, the resist is exposed to light using a photomask on which a pattern corresponding to the migration channel 30 is formed, and is further developed to form a resist layer 70. Thereafter, the substrate is washed with pure water and dried with a spin drier.
[0034]
Next, as shown in FIG. 2B, the exposed substrate 20 is etched using the resist layer 70 as a mask, and a plurality of grooves 22 are formed on the first main surface 20a side in a predetermined manner. Form at intervals. This etching step is preferably performed by, for example, reactive ion etching (hereinafter, also simply referred to as RIE) using oxygen plasma. As the reaction conditions, conventionally known reaction conditions can be applied. However, the groove 22 having a predetermined shape is formed by appropriately adjusting the reaction conditions in consideration of the width, depth, and cross-sectional shape of the groove 22. can do.
[0035]
In the example of the manufacturing method of this embodiment, the example in which the groove 22 is formed by performing reactive ion etching has been described. However, the groove 22 may be mechanically formed by, for example, a so-called slicing device (dicing device).
[0036]
Next, as shown in FIG. 2C, using the patterned resist layer 70 as a mask, the migration channel 30 is formed so as to fill the groove 22. The migration channel 30 is formed of a porous material having a pore diameter and a porosity large enough to allow electrophoresis. Preferably, the material is made of, for example, porous silica.
[0037]
Here, a step of forming the migration channel 30 will be described. The steps (1) to (4) described below are steps for forming a migration channel 30 suitable for application to the electrophoresis device 10 of the present invention.
[0038]
▲ 1 ▼ First method
The first method is a forming step by a so-called sol-gel method. First, a mixed solution containing an alkoxide, an alcohol as a solvent, an acid or base as a catalyst, and water for hydrolysis is used as a starting solution, and the mixed solution is heated to a predetermined temperature to perform hydrolysis and polymerization. A sol-like solution is obtained by the condensation reaction. This sol-like solution is applied by, for example, a conventionally known spin coating method so as to fill the groove 22 via the resist layer 70. Thereafter, by heating at a predetermined temperature and time, water and the solvent are evaporated to form a porous body. Preferably, for example, a methoxide silicate is used as an alkoxide, and a molar ratio of methoxide silicate: water: methanol: hydrogen chloride = 1: 4: 2: 0.4 is used as a spin coating method. Is applied on the resist layer 70 so as to fill the groove 22. Thereafter, it is preferable to form the electrophoresis channel 30 by heating at a predetermined temperature and time to evaporate water and the solvent.
[0039]
For example, as a specific reaction condition, there is an example in which baking is performed at 150 ° C. for 1 hour. Thus, a migration channel having an average pore diameter of 3 nm and a porosity of 0.6% is obtained. The “average pore diameter” and “porosity” here can be determined as approximate values estimated from images of the cross section of the migration channel by a transmission electron microscope.
[0040]
In general, the range of the pore diameter suitable for protein electrophoresis is 1 nm to 10 μm, preferably 2 nm to 50 nm. The porosity is preferably in the range of 0.5% to 50%.
[0041]
In this sol-gel method, for example, Al ₂ O ₃ , MgO, SiO ₂ , ZrO ₂ And TiO ₂ The inorganic substance is selected from any of the above. Further, for example, an organic substance is selected from any of organic compounds having an alkyl group, an acyl group or a phenyl group. Then, an organic-inorganic composite obtained by chemically bonding them can be used.
[0042]
For example, a mixture of any one of alkylalkoxysilanes selected from the group including methyltriethoxysilane and dimethyldiethoxysilane, and an alkoxysilane selected from the group including tetraethoxysilane, or a polydiphenylsiloxane oligomer and tetraethoxysilane After forming a film of the mixture on the substrate 20, the organic component is removed by heating to form a porous body.
[0043]
Specifically, a sol-gel film can be formed using methyldiethoxysilane and tetraethoxysilane. In this case, an organic-inorganic composite film in which a methyl group remains can be formed. Also, a sol-gel film can be formed using a polydiphenylsiloxane oligomer and tetraethoxysilane. In this case, an organic-inorganic composite film in which phenyl groups remain can be formed.
[0044]
(2) Second method
The second method uses a spin-on-glass solution (for example, SiO2 manufactured by Tokyo Ohka Co., Ltd.). ₂ In this step, the electrophoresis channel 30 is formed using a coating liquid for forming a film, such as OCD-T10, or a dispersion of ultrafine silica particles.
[0045]
For example, iodine is added to the above-mentioned spin-on-glass solution, and this iodine-containing solution is buried in the groove 22 by using a pattern of the resist layer 70 by a spin coating method. After the iodine is gradually desorbed, a heat treatment is performed, for example, at 450 ° C. for 1 hour in a nitrogen atmosphere, whereby the migration channel 30 having a pore diameter of about 60 nm can be formed.
[0046]
(3) Third method
The third method is a method of forming a porous silicon layer having a pore diameter of about 10 nm by performing the same anodic oxidation treatment of a silicon substrate as in JP-A-9-304340 described in the section of the prior art.
[0047]
▲ 4 ▼ Other methods
Other methods include applicable, for example, zeolite or Al ₂ O ₃ , MgO, SiO ₂ , ZrO ₂ Or TiO ₂ A porous film can be formed by depositing such an oxide on a substrate by a conventionally known sputtering process. At this time, the pore diameter can be controlled by adjusting the sputtering conditions.
[0048]
Furthermore, using a polymer material such as polyamide, polyacetal, polycarbonate, and polyacrylamide, it is possible to form a porous film by a conventionally known blowing agent decomposition method, a solvent diffusion method, a chemical reaction method, or a firing method. It is possible.
[0049]
Preferably, the migration channel 30 is formed by the sol-gel method (1) in consideration of ease of implementation.
[0050]
Next, as shown in FIG. 3A, the resist layer 70 is removed by using an appropriate stripper for stripping the resist.
[0051]
After that, as shown in FIG. 3B, the exposed upper surface of the migration channel 30 is polished and flattened by, for example, a CMP (chemical mechanical polishing) method. At this time, it is preferable to flatten the surface so that the level (that is, the height, hereinafter the same) of the upper surface of the migration channel and the level of the first main surface 20a of the substrate 20 are substantially the same. . However, the upper surface of the migration channel 30 may protrude at a higher level than the first main surface 20a. When the upper surface of the electrophoresis channel 30 can be formed uniformly, that is, the cross-sectional and vertical cross-sectional shapes of the electrophoresis channel 30 can be formed uniformly over the entire length of the electrophoresis channel 30 so as not to hinder electrophoresis. This polishing step is not always necessary. In this case, the manufacturing process can be further simplified.
[0052]
Next, in order to protect the formed migration channels 30, a channel protective film 40 is formed on the first main surface of the substrate 20 so as to cover the migration channels 30.
[0053]
The electrophoresis apparatus 10 of this embodiment is assumed to obtain an analysis result from the first main surface 20a side of the substrate 20, that is, to analyze a migration pattern of a sample. Therefore, the channel protective film 40 is preferably formed by selecting an appropriate material within a range that does not impair the resolution in the analysis. Preferably, this protective film 40 is made of SiO ₂ It is good to be a membrane. This protective film 40 is made of SiO ₂ When the film is formed by, for example, a thin film having a thickness of about 1 μm to 3 μm is preferable.
[0054]
The formation of such a channel protective film 40 is performed, for example, by using SiO 2 ₂ In the case of a film, a conventionally known general condition is applied, and a plasma CVD method using an ethyl silicate (TEOS) gas is used. ₂ Can be deposited in a desired thickness.
[0055]
Here, with reference to FIGS. 4A to 4C, a process of forming the exposed first and second edge portions 32a and 32b of the migration channel 30 will be described.
[0056]
FIGS. 4A to 4C are schematic cross-sectional views each showing a part of a cut surface taken along line II ′ of FIG. 1A. This sectional view is a longitudinal sectional view along the extending direction of the migration channel 30. Only the vicinity of the edge portion 32 is shown in an enlarged manner, and other portions are omitted.
[0057]
As shown in FIG. 4A, the migration channel 30 is formed on the first main surface 20a side of the substrate 20 by the above-described process.
[0058]
Thereafter, as shown in FIG. 4B, the edges 32 of the electrophoresis channel, that is, the regions to be the first and second edges 32a and 32b are covered by the edge-exposing protective film 80. Cover it. As the protective film 80 for exposing the edge portion, a metal mask made of, for example, a stainless alloy is preferably arranged in contact, or a film of a heat resistant resin such as a photopolymerizable polyimide is preferably formed by a thick film printing method.
[0059]
After the formation of the channel protective film 40, as shown in FIG. 4C, the protective film 80 for exposing the edge portion is removed by peeling, if necessary, by a peeling means such as an appropriate peeling liquid. I do. In this manner, the first and second edge portions 32a and 32b are exposed and formed.
[0060]
Thereafter, the plurality of structures (electrophoresis devices) formed in a lattice shape on the substrate 20 are cut into individual pieces by cutting between adjacent structures. In this way, a plurality of electrophoresis devices 10 are obtained from one large-sized substrate. This singulation step is preferably performed by, for example, a dicing apparatus using a high-speed rotating blade.
[0061]
According to the method of manufacturing the electrophoresis device of the first embodiment, a large number of electrophoresis devices of the first embodiment can be efficiently manufactured simultaneously with simple steps.
[0062]
(Second embodiment)
With reference to FIGS. 5A, 5B, and 5C, a configuration of the electrophoresis apparatus according to the second embodiment of the present invention will be described. The components already described in the first embodiment are given the same reference numerals and their detailed description is omitted. In addition, since the materials applied, the execution conditions of each step, and the like can be the same as those described in the first embodiment, detailed descriptions thereof may be omitted.
[0063]
FIG. 5A is a schematic plan view of the configuration of the electrophoresis apparatus according to the second embodiment as viewed from above, and FIG. 5B is a cross-sectional view taken along line II of FIG. FIG. 5C is a schematic cross-sectional view showing a cut surface taken along a line, and FIG. 5C is a schematic sectional view showing a cut surface taken along a dashed line II-II of FIG. It is sectional drawing.
[0064]
The electrophoresis apparatus 10 according to the second embodiment includes a substrate 20. The substrate 20 has a first main surface 20a and a second main surface 20b facing the first main surface 20a.
[0065]
The substrate 20 is a plate-shaped body in which the first main surface 20a and the second main surface 20b have the same shape and the same rectangular shape.
[0066]
On the first main surface 20a of the substrate 20, a plurality of cylindrical electrophoresis channels 30 having a rectangular cross section are formed linearly. These migration channels 30 are formed parallel to each other and apart from each other along the longitudinal direction of the first main surface 20a. These migration channels 30 are defined as linear first migration channels 30-1, second migration channels 30-2, third migration channels 30-3,..., And n-th migration channels 30-n ( n is a positive number of 4 or more.).
[0067]
The migration channel 30 is formed of a porous body.
[0068]
The first and second edges 32a and 32b, ie, both ends 32 of the migration channel 30, are exposed.
[0069]
The surface of the migration channel 30 is covered with a protective film 34.
[0070]
The migration channel protection film 40 is formed on the substrate 20 on which the migration channel and the protection film 34 are provided so as not to cover both end portions of the first main surface 20a in the longitudinal direction. Therefore, the first and second edges 32a and 32b of the migration channel 30 are exposed.
[0071]
With the configuration of the electrophoresis apparatus according to the second embodiment, the same operation and effect as those of the electrophoresis apparatus according to the first embodiment can be obtained.
[0072]
Next, a method for manufacturing the electrophoresis device according to the second embodiment will be described with reference to FIGS.
[0073]
6 (A) to 6 (D) are schematic cross-sectional views showing a cut surface taken along a line II-II ′ of FIG. 5 (A).
[0074]
A region for forming a plurality of electrophoresis devices 10 is set in advance on a large-sized substrate 20 (not shown). Then, in this region, the spacing between the semiconductor devices 10, the shape and width of the migration channels 30, and the spacing between the adjacent migration channels 30 are set assuming that the semiconductor devices 10 will be separated later.
[0075]
Next, as shown in FIG. 6A, a porous film for forming the migration channel 30, that is, a film 30 for forming the migration channel, is formed on the entire surface of the first main surface 20a of the substrate 20. Is formed with a thickness equal to a predetermined thickness of the migration channel 30 which is set in advance. The step of forming the migration channel forming film 30 'is preferably performed by any one of the steps (1) to (4) already described in the first embodiment.
[0076]
Then, as shown in FIG. 6B, for example, a film 30 ′ for forming an electrophoretic channel is formed by a plasma CVD method using an ethyl silicate (TEOS) gas by applying a conventionally known general condition. SiO for protection ₂ Is formed on the entire surface of the film 30 '. The protective film 34 may have such a thickness that the film 30 ′ for forming the migration channel can be protected from the resist layer 70 formed later. The thickness of the protective film 34 is preferably, for example, about 0.2 μm.
[0077]
Next, as shown in FIG. 6B, a resist layer 70 is formed by a photolithography process under the same conditions as those applied in a conventionally known semiconductor device manufacturing process. As a material for the resist layer 70, a silicon-containing positive resist is preferably used, and spin coating, prebaking, photomask formation, exposure and development steps are performed to form the resist layer 70. The resist layer 70 is patterned so as to cover a channel forming film 30 ′ to be the migration channel 30 to be formed.
[0078]
Next, as shown in FIG. 6C, using the patterned resist layer, that is, the resist pattern 70 as a mask, a channel forming film 30 ′ exposed in a region where the resist pattern 70 is not formed. The migration channel 30 is formed by patterning by performing an etching process on. This etching step is preferably performed by, for example, reactive ion etching (RIE) using oxygen plasma. As the reaction conditions, conventionally known reaction conditions can be applied. In particular, by appropriately adjusting the reaction conditions in consideration of the predetermined width, depth and cross-sectional shape of the migration channel 30, the electrophoresis having the predetermined shape can be performed. A channel 30 can be formed. The electrophoresis channel 30 is preferably made of a porous material having a pore diameter and a porosity large enough to allow electrophoresis as described above, for example, porous silica.
[0079]
Next, the resist layer 70 is removed with a suitable stripper for stripping the resist.
[0080]
Next, as shown in FIG. 6D, in order to protect the plurality of migration channels 30 formed, a channel protection film 40 is formed on the first main surface of the substrate 20 so as to cover the migration channels 30. Formed.
[0081]
In the manufacturing method according to the second embodiment, a configuration example will be described in which, for example, a fluorinated polyimide is used as the channel protective film 40 in consideration of the analysis accuracy of the migration result and the like. It is more preferable that the channel protective film 40 is formed of fluorinated polyimide, because light absorption in the visible light region can be suppressed. Further, the protective film 40 can be formed as a thin film having a desired thickness by a simpler process such as a coating method. The channel protective film 40 is preferably formed as a thin film having a thickness of about 10 μm.
[0082]
Such a fluorinated polyimide layer as the channel protective film 40 can be formed with a desired thickness under general conditions by applying a conventionally known coating method or the like, for example.
[0083]
Here, with reference to FIGS. 7A and 7B, a process of forming the exposed first and second edge portions 32a and 32b of the migration channel 30 will be described.
[0084]
FIGS. 7A and 7B are schematic cross-sectional views showing a cut surface taken along the line II ′ of FIG. 5A. The edge portion 32, that is, the vicinity of the first and second edge portions 32a or 32b is shown in an enlarged manner, and other portions are omitted.
[0085]
As shown in FIG. 7A, the migration channel 30 is formed on the first main surface 20a side of the substrate 20 by the above-described process.
[0086]
Then, as shown in FIG. 7B, the channel protective film 40, which is a fluorinated polyimide resin layer, and the protective film 34 under the channel protective film 40 are partially removed, and the edge 32 of the migration channel 30 is removed. Expose. The channel protective film 40 and the protective film 34 are partially removed by, for example, using a slicing device to cut off only a desired edge region.
[0087]
Thereafter, the plurality of structures formed in a lattice on the substrate 20 are cut into individual pieces by cutting between adjacent structures. Thus, a plurality of electrophoresis devices 10 are obtained from one large-sized substrate 20.
[0088]
As described above, according to the method of manufacturing the electrophoresis apparatus of the second embodiment, a large number of electrophoresis apparatuses of the second embodiment can be efficiently manufactured with simple steps.
[0089]
In particular, in the formation of the migration channel, since the patterning can be performed after the material is formed as a film on the entire surface of the substrate, a plurality of migration channels can be manufactured more uniformly.
[0090]
(Third embodiment)
With reference to FIGS. 8A, 8B, and 8C, a configuration of the electrophoresis apparatus according to the third embodiment of the present invention will be described. The components already described in the first and second embodiments are denoted by the same reference numerals, and detailed description thereof will be omitted. In addition, as for the applied material, the process conditions, and the like, those described in the first and second embodiments can be applied in the same manner, and thus detailed description thereof is omitted.
[0091]
FIG. 8A is a schematic plan view illustrating the configuration of the electrophoresis apparatus according to the third embodiment as viewed from above, and FIG. 8B is a view taken along the line II of FIG. FIG. 8C is a schematic cross-sectional view illustrating a vertical cross section taken along a broken line, and FIG. 8C is a schematic cross-sectional view illustrating a cross section taken along a II-II ′ broken line in FIG. It is sectional drawing.
[0092]
The electrophoresis apparatus 10 according to the third embodiment includes a substrate 20. The substrate 20 has a first main surface 20a and a second main surface 20b facing the first main surface 20a.
[0093]
The substrate 20 is a plate-like body in which the first main surface 20a and the second main surface 20b have the same shape and the same rectangular shape.
[0094]
A plurality of grooves 22 are formed on the first main surface 20 a of the substrate 20. The groove 22 has a rectangular cross section in this configuration example. These grooves 22 are formed in a linear shape along the longitudinal direction of the first main surface 20a, parallel to each other, and separated from each other.
[0095]
An electrophoresis channel 30 is formed in each of the plurality of grooves 22. The migration channel 30 is formed by embedding the groove 22 with a porous material.
[0096]
Therefore, the linear migration channels 30 are formed parallel to each other and separated from each other along the longitudinal direction of the first main surface 20a.
[0097]
On the first main surface 20a of the substrate 20 where the migration channel 30 is formed, both ends 32 of the migration channel 30, that is, the first and second edge portions 32a and 32b are exposed. A protective film 40 is formed.
[0098]
A first electrode 50 is formed of a conductive material on the first edge 32a of the migration channel 30. A second electrode 52 is formed of a conductive material on the second edge 32b. The first and second electrodes 50 and 52 are respectively formed in the first to n-th migration channels 30-1 to 30-n. The first and second electrodes 50 and 52 are preferably formed by forming, for example, a chromium (Cr) thin film and then forming a gold (Au) thin film on the chromium thin film.
[0099]
The first and second electrodes 50 and 52 formed in the migration channel 30 of the electrophoresis apparatus 10 according to the third embodiment are respectively composed of the first migration channel 30-1 to the n-th migration channel. Although the configuration example formed in each of the 30-n has been described, the first and second electrodes 50 and 52 may be formed so as to extend over the plurality of migration channels 30. For example, the first electrode 50 may be formed as one electrode so as to extend over all the migration channels 30 on the side of the first edge 32 a of the migration channel 30.
[0100]
Further, the electrophoretic device 10 according to the third embodiment is characterized in that an opening 42 is formed in this configuration example. The opening 42 is opened from the surface of the channel protective film 40 to the surface of the migration channel 30 in order to supply a sample to be subjected to electrophoresis to each migration channel 30. In this example, one opening 42 is formed for each migration channel 30.
[0101]
The shape and size of the opening 42 can be determined within a range that does not hinder the supply of the sample.
[0102]
The opening 42 extends over the plurality of migration channels 30, that is, in the direction perpendicular to the direction in which the migration channels 30 extend, as one or more openings 42, for example, in which the plurality of migration channels 30 It may be configured to be exposed by the opening 42.
[0103]
The opening 42 can be formed at any suitable position on the migration channel 30 between the first electrode 50 and the second electrode 52 of each migration channel 30.
[0104]
In particular, as shown in FIG. 8A, when the opening 42 is provided closer to the center between the first electrode 50 and the second electrode 552, the sample is moved according to the charge of the sample. Electrophoresis can be performed from the opening 42 to the first electrode 50 side and from the opening 42 to the second electrode 52 side, that is, in both directions in which the migration channel 30 extends. If electrophoresis in a single direction is desired, the opening 42 may be formed near the first electrode 50 or the second electrode 52.
[0105]
In the electrophoresis apparatus 10 according to this embodiment, the configuration example in which both the first and second electrodes 50 and 52 and the opening 42 are provided has been described, but a configuration in which only one of them is provided may be employed. it can.
[0106]
Note that the electrophoresis apparatus 10 of this embodiment shows a configuration example in which first and second electrophoresis buffer supply means 60 and 62 are added. The first and second electrophoresis buffer supply means 60 and 62 are configured to supply a buffer for electrophoresis to the electrophoresis channel 30 and to derive the electrophoresis buffer from the electrophoresis channel 30. . These are preferably made of, for example, fiber fibers of the same material and the same shape. A specific electrophoresis process will be described later.
[0107]
According to the configuration of the electrophoresis apparatus of the third embodiment, in addition to the same operation and effect as the electrophoresis apparatus of the first embodiment, the first and second electrodes 50 and 52 are When each is formed in each of the first migration channel 30-1 to the n-th migration channel 30-n, electrophoresis is performed independently by applying an electric field to each migration channel 30 individually. Therefore, the electrophoresis apparatus 10 can be operated flexibly. Further, since the opening 42 is formed, more accurate supply of the sample to the migration channel 30 can be performed.
[0108]
Next, a method of manufacturing the electrophoresis device according to the third embodiment will be described with reference to FIGS.
[0109]
FIGS. 9A to 9C are schematic cross-sectional views showing a cross section taken along the line II-II ′ of FIG. 8A.
[0110]
First, regions for forming a plurality of electrophoresis devices 10 are set in advance on the substrate 20 (not shown). Then, in this region, the spacing between the semiconductor devices 10, the shape and width of the migration channels 30, and the spacing between the adjacent migration channels 30 are set assuming that the semiconductor devices 10 will be separated later.
[0111]
As shown in FIG. 9A, the migration channel 30 is formed by the same steps as those already described in the first embodiment. The migration channel 30 may be formed by any of the above methods (1) to (4).
[0112]
Thereafter, as shown in FIG. 9B, the exposed upper surface of the migration channel 30 is polished and flattened by, for example, a CMP (chemical mechanical polishing) method.
[0113]
Next, in order to protect the formed plurality of migration channels 30, a channel protection film 40 is formed on the first main surface 20 a of the substrate 20 so as to cover the migration channels 30.
[0114]
Next, as shown in FIG. 9C, a resist layer 70 having a pattern exposing a region where the opening 42 is formed is formed by a conventionally known photolithography method. Next, using the resist layer 70 as a mask, the opening 42 is formed by reactive ion etching using, for example, oxygen plasma as described above so that a part of each of the plurality of migration channels 30 is exposed.
[0115]
Here, with reference to FIGS. 10A to 10D, the exposed first and second edge portions 32a and 32b of the migration channel 30 and the first and second electrodes 50 and 52. Will be described. These steps are performed in parallel with the steps described with reference to FIGS. 9A to 9C described above.
[0116]
FIGS. 10A to 10D are schematic cross-sectional views showing a vertical cross section taken along the line II ′ of FIG. 8A. Only the vicinity of the edge portion 32 is shown in an enlarged manner, and other portions are omitted.
[0117]
As shown in FIG. 10A, the migration channel 30 is formed on the first main surface 20a side of the substrate 20 by the above-described process.
[0118]
Further, as shown in FIG. 10B, a migration channel protective film 40 is formed. Next, a resist layer (that is, a resist pattern) 70 having a pattern in which the edge 32 of the electrophoresis channel, that is, the region that becomes the first and second edge 32a and 32b and the region that becomes the opening 42 is exposed, It is formed by a conventionally known photolithography method.
[0119]
Next, as shown in FIG. 10C, the above-described reactive ion etching using, for example, oxygen plasma is performed using the resist layer 70 as a mask so that a part of the surface of the migration channel 30 is exposed. Thus, the first and second edge portions 32a and 32b and the opening 42 are formed.
[0120]
Thereafter, as shown in FIG. 10D, the first and second electrodes 50 and 52 are formed on the exposed edge 32 using a conductive material. Preferably, for example, using a mask having a configuration in which only the regions where the electrodes 50 and 52 are to be formed is exposed, using a metal mask such as a stainless alloy, or the like, and using a conventionally known electron beam evaporation method, firstly, chromium (Cr). The first and second electrodes 50 and 52 can be formed by forming a thin film of about 10 nm and a thin film of gold (Au) on this chromium thin film with a thickness of about 500 nm.
[0121]
After that, the plurality of structures (electrophoresis devices) formed in a lattice on the substrate 20 are cut into individual pieces by cutting between adjacent structures. In this way, a plurality of electrophoresis devices 10 are obtained from one large-sized substrate.
[0122]
As described above, according to the method of manufacturing the electrophoresis apparatus of the third embodiment, a large number of electrophoresis apparatuses of this embodiment can be efficiently manufactured with simple steps.
[0123]
(Fourth embodiment)
With reference to FIGS. 11A, 11B and 11C, a configuration of an electrophoresis apparatus according to a fourth embodiment of the present invention will be described. Note that the components already described in the first to third embodiments are given the same reference numerals and may not be described in detail. In addition, since the applied materials, the conditions for implementing each step, and the like can be applied in principle to the examples described in the first to third embodiments, detailed descriptions thereof are omitted.
[0124]
FIG. 11A is a schematic plan view of the configuration of the electrophoresis apparatus according to the fourth embodiment as viewed from above, and FIG. 11B is a cross-sectional view taken along line II of FIG. 11A. FIG. 11C is a schematic cross-sectional view showing a cut surface cut along a broken line, and FIG. 11C is a schematic cross-sectional view showing a cut surface cut along a II-II ′ broken line in FIG. 11A. It is sectional drawing.
[0125]
The electrophoresis apparatus 10 according to the fourth embodiment includes a substrate 20. The substrate 20 has a first main surface 20a and a second main surface 20b facing the first main surface 20a.
[0126]
The substrate 20 is a plate-like body in which the first main surface 20a and the second main surface 20b have the same shape and the same rectangular shape.
[0127]
On the first main surface 20a of the substrate 20, a plurality of cylindrical electrophoresis channels 30 having a rectangular cross section are formed linearly.
[0128]
The migration channel 30 is formed of a porous body. Both ends 32 of the electrophoresis channel 30, that is, first and second edges 32a and 32b are exposed.
[0129]
A first electrode 50 is formed of a conductive material on the first edge 32a of the migration channel 30. A second electrode 52 is formed of a conductive material on the second edge 32b. The first and second electrodes 50 and 52 are respectively formed in the first to n-th migration channels 30-1 to 30-n.
[0130]
The surface of the migration channel 30 is covered with a protective film 34.
[0131]
The migration channel protection film 40 is formed on the substrate 20 on which the migration channel and the protection film 34 are provided so that the first and second edge portions 32a and 32b are exposed.
[0132]
Further, the configuration example of the electrophoresis apparatus 10 according to the fourth embodiment is characterized in that an opening 42 is formed. The opening 42 is opened from the surface of the channel protective film 40 to the surface of the migration channel 30 in order to supply a sample to be subjected to electrophoresis to each migration channel 30. In this example, one opening 42 is formed for each migration channel 30.
[0133]
The shape and size of the opening 42 can be determined within a range that does not hinder the supply of the sample.
[0134]
The opening 42 can be formed so as to extend over the plurality of migration channels 30 as already described in the third embodiment.
[0135]
The opening 42 can be formed at any suitable position on the migration channel 30 between the first electrode 50 and the second electrode 52 of each migration channel 30.
[0136]
In particular, as shown in FIG. 11A, when the opening 42 is provided closer to the center between the first electrode 50 and the second electrode 52, the sample is moved according to the charge of the sample. Electrophoresis can be performed from the opening 42 toward the first electrode 50 and from the opening 42 toward the second electrode 52 in both directions in which the migration channel 30 extends. If electrophoresis in a single direction is desired, the opening 42 may be formed near the first electrode 50 or the second electrode 52.
[0137]
In the electrophoresis apparatus 10 according to this embodiment, the configuration example in which both the first and second electrodes 50 and 52 and the opening 42 are provided has been described, but a configuration in which only one of them is provided may be employed. it can.
[0138]
According to the configuration of the electrophoresis apparatus of the fourth embodiment, in particular, the first and second electrodes 50 and 52 respectively include the first migration channel 30-1 to the n-th migration channel 30-n. In this case, the electrophoresis apparatus 10 can be operated flexibly because an electric field can be individually applied to each migration channel 30 to perform electrophoresis independently of each other. . Further, since the opening 42 is formed, more accurate supply of the sample to the migration channel 30 can be performed.
[0139]
Next, a method for manufacturing an electrophoresis device according to a fourth embodiment will be described with reference to FIGS.
[0140]
FIGS. 12A to 12D are schematic cross-sectional views showing a vertical cross section taken along line II ′ of FIG. 11A. Only the vicinity of the edge portion 32 is shown in an enlarged manner, and other portions are omitted.
[0141]
First, regions for forming a plurality of electrophoresis devices 10 are set in advance on the substrate 20 (not shown). Then, in this region, the spacing between the semiconductor devices 10, the shape and width of the migration channels 30, and the spacing between the adjacent migration channels 30 are set assuming that the semiconductor devices 10 will be separated later.
[0142]
As shown in FIG. 12A, the migration channel 30 is formed by the steps already described in the second embodiment. The migration channel 30 (including the film before patterning) may be formed by any of the methods (1) to (4) described in the first embodiment. Next, in order to protect the formed plurality of migration channels 30, a channel protection film 40 is formed on the first main surface 20 a of the substrate 20 so as to cover the migration channels 30.
[0143]
Next, as shown in FIG. 12B, a pattern in which the edge 32 of the electrophoresis channel, that is, the regions that become the first and second edges 32a and 32b and the region that becomes the opening 42 are exposed is formed. The resist layer 70 to be formed is formed by a conventionally known photolithography method.
[0144]
As shown in FIG. 12C, etching of the migration channel protective film 40 and the protective film 34 therebelow are performed by the above-described reactive ion etching using, for example, oxygen plasma using the resist layer 70 as a mask. Do. This etching forms first and second edges 32a and 32b and an opening 42, respectively, exposing a portion of the surface of the migration channel 30.
[0145]
Next, the first and second electrodes 50 and 52 are formed on the exposed edge 32 using a conductive material. Preferably, for example, a metal mask made of a stainless alloy or the like, which is configured to expose only the regions where the electrodes 50 and 52 are to be formed, is formed. Thereafter, using this metal mask, a conventionally known electron beam evaporation method is used. First, a chromium (Cr) thin film is formed with a thickness of about 10 nm. Thereafter, a gold (Au) thin film may be formed on the chromium thin film to a thickness of about 500 nm.
[0146]
Thereafter, the plurality of structures (electrophoresis devices) formed in a lattice shape on the substrate 20 are cut into individual pieces by cutting between adjacent structures. In this way, a plurality of electrophoresis devices 10 are obtained from one large-sized substrate.
[0147]
According to the method of manufacturing the electrophoresis device of the fourth embodiment, a large number of electrophoresis devices of the fourth embodiment can be efficiently manufactured with simple steps.
[0148]
(Fifth embodiment)
With reference to FIGS. 13A and 13B, a configuration of an electrophoresis apparatus according to a fifth embodiment of the present invention will be described. The components already described in the first to fourth embodiments are denoted by the same reference numerals, and detailed description thereof will be omitted. Further, the applied materials and the execution conditions of each step can be applied in principle to the examples described in the first to fourth embodiments, and therefore, detailed description thereof is omitted.
[0149]
FIGS. 13A and 13B are schematic plan views illustrating the configuration of the electrophoresis apparatus according to the fifth embodiment, as viewed from above. FIG. 13A shows a first configuration example. FIG. 13B shows a second configuration example.
[0150]
The electrophoresis apparatus 10 according to the fifth embodiment is characterized by the shape of the migration channel 30. The configuration example of the electrophoresis channel 30 of the electrophoresis apparatus 10 according to the fifth embodiment can be applied to any of the electrophoresis apparatuses described in the first to fourth embodiments. Therefore, detailed description of the configuration other than the migration channel 30 is omitted.
[0151]
The electrophoresis apparatus 10 includes a substrate 20. The substrate 20 has a plurality of migration channels 30 formed therein. These migration channels 30 are formed apart from each other. The migration channel 30 is formed of a porous body.
[0152]
As shown in FIG. 13A, in the first configuration example, the plurality of migration channels 30 extend so that the plurality of curved portions 30 b are sequentially connected in the longitudinal direction of the migration device 10. That is, the migration channel 30 is formed in a corrugated shape obtained by combining the curved portions 30b on the flat substrate upper surface. The curved portion 30b is configured as, for example, a part of a circular arc or an elliptical arc. The portions of the arc can be combined in any suitable range and size.
[0153]
As shown in FIG. 13B, in the second configuration example, the plurality of migration channels 30 include the plurality of linear portions 30a formed in parallel with each other. A predetermined number of the linear portions 30a, in this configuration example, three are set as one set, and the ends of the linear electrophoresis channels adjacent on the same side in this set are connected by a curved portion 30c. Therefore, the migration channel 30 of this configuration example has a folded structure that connects a plurality of linear channels to each other.
[0154]
In the first and second configuration examples, in particular, the migration channel 30 near the first and second edge portions 32a and 32b considers the arrangement of the opening 42 and the first and second electrodes 50 and 52. And it is linear.
[0155]
On the first main surface 20 of the substrate 20 on which the migration channel 30 is formed, a migration channel protective film 40 is formed.
[0156]
An opening 42 is formed in the channel protective film 40. The opening 42 is open from the surface of the channel protective film 40 to the surface of the migration channel 30. In this embodiment, one opening 42 is formed for each migration channel 30.
[0157]
The opening 42 can be formed at any suitable position on the migration channel 30 between the first electrode 50 and the second electrode 52 of each migration channel 30.
[0158]
First and second electrodes 50 and 52 are formed on the exposed first and second edge portions 32a and 32b.
[0159]
In the electrophoresis apparatus 10 according to this embodiment, the example in which the opening 42 and the first and second electrodes 50 and 52 are formed in the linear portion of the migration channel 30 has been described. However, the present invention is not limited thereto. For example, it can be formed on the curved portion 30b.
[0160]
Further, although an example has been described in which the electrophoresis channel 30 of the present embodiment extends in the longitudinal direction of the electrophoresis apparatus 10, an arbitrary suitable shape can be used as long as the object of the present invention is not impaired. The migration channel 30 may be formed in a spiral shape, for example. For example, once draw a spiral in a specific direction toward the center of the circle, gradually draw a small circle, turn around near the center, and then follow the spiral already drawn, gradually in the opposite direction A channel shape that draws a spiral like a large circle may be used.
[0161]
Furthermore, an example has been described in which the migration channel 30 is configured to pass from the first edge portion 32a to the second edge portion 32b side. For example, the migration channel 30 is formed in a curved shape by the curved portion as described above. In such a case, the end of the electrophoresis channel 30 may be shaped to pass through to the same side. That is, the end of the migration channel 30 starting from the first edge portion 32a side may be set to the first edge portion 32a side.
[0162]
According to the configuration of the electrophoresis apparatus of the fifth embodiment, the length of the migration channel 30 can be made longer, so that the resolution in analysis can be improved.
[0163]
The manufacture of the electrophoresis apparatus according to the fifth embodiment can be carried out as already described in the above-described first to fourth embodiments. A preferred mode of use of the electrophoresis apparatus, in particular, a protein electrophoresis method will be described.
[0164]
First, the two-dimensional electrophoresis will be described. The so-called two-dimensional electrophoresis method includes a two-stage electrophoresis method focusing on different properties of a protein. For example, as the first dimension electrophoresis, isoelectric focusing using the isoelectric point of a protein as an index, and as the second dimension electrophoresis, SDS-polyacrylamide gel electrophoresis using the molecular weight of a protein as an index (Hereinafter, also simply referred to as SDS-PAGE).
[0165]
Isoelectric focusing uses an immobilized pH gradient gel that is generally formed to have a longitudinal pH gradient. When electrophoresis is performed using such a pH gradient gel, the protein molecule stays at the position of pH corresponding to the intrinsic charge of the protein molecule because the pH of the gel is equal to the potential of the protein. In this manner, a protein having a specific charge can be separated from a mixture of a plurality of types of proteins using the charge of the protein as an index.
[0166]
In addition, SDS-PAGE refers to performing electrophoresis in a polyacrylamide gel using a sample in which a protein is denatured in a chain with a treatment solution containing sodium dodecyl sulfate (hereinafter, also simply referred to as SDS). This is an electrophoresis method. The composition of the processing solution and the buffer for electrophoresis, and the electrophoresis conditions are described in, for example, publications and standard academic books (for example, Laboratory Chemistry Course, Maruzen, etc.). Conditions can be selected.
[0167]
In a gel, the larger the molecule, the more difficult it is to move, so the longer the chain length of the denatured protein, the lower the mobility. In this way, a protein having a specific molecular weight can be separated from a mixture of a plurality of types of proteins using the molecular weight (chain length) of the protein as an index.
[0168]
In the two-dimensional electrophoresis method, generally, first, after isoelectric focusing, SDS-PAGE is continuously performed.
[0169]
Next, an electrophoresis process using the semiconductor device of the present invention will be schematically described.
[0170]
First, an electrophoresis buffer is introduced into the electrophoresis channel 30 of the electrophoresis apparatus 10 according to the above-described embodiment. As the electrophoresis buffer, a buffer having any suitable composition (for example, for SDS-PAGE) well known to those skilled in the art based on the publications and standard academic books described above can be used.
[0171]
Two methods can be applied for introducing this buffer. First, the edge 32 of the electrophoresis channel 30, that is, one of the first edge 32 a and the second edge 32 b is directly immersed in the container containing the buffer for electrophoresis. In this method, an electrophoresis buffer is injected into the electrophoresis channel 30 by capillary action. Second, using a fibrous fiber, one end of the fibrous fiber is immersed in a container containing a buffer solution for electrophoresis, and the other end is connected to the first edge 32a or the second end. This is a method of injecting the electrophoresis buffer into the electrophoresis channel 30 by capillary action by connecting to one of the edges 32b.
[0172]
In this way, the electrophoresis apparatus 10 into which the electrophoresis buffer has been injected can be stored in a moist state for a certain period of time without being immediately subjected to electrophoresis.
[0173]
Next, a sample to be subjected to electrophoresis will be described. Examples of the electrophoresis sample include SDS mainly applied to SDS-PAGE, for example, a sample treated with a buffer having any suitable composition and processing conditions known to those skilled in the art according to publications and standard academic books. Is assumed to be applied. However, as will be described later, as the first dimension electrophoresis, the sample at the stage where the isoelectric focusing as described above has been completed is not subjected to further processing (for example, the above-described processing using a buffer containing SDS), It can be directly subjected to the second-dimensional electrophoresis.
[0174]
The sample to be subjected to electrophoresis can be analyzed using the generation of fluorescence by excitation light as an index without performing staining, but in order to further improve the resolution of the analysis, for example, the sample is previously stained with a fluorescent reagent. Good to put. Specific examples of the fluorescent reagent include (i) fluorescamine which reacts with a primary amine, and (ii) a BODIPY (registered trademark) which covalently binds to an n-terminal amino group of a side chain of a protein or an ε-amino group of a lysine residue. ) TR-X dye, (iii) and others such as fluorescein isocyanate (FITC), phycoerythrin (PE) and periodinin chlorophyll protein (PerCP).
[0175]
The sample to be subjected to electrophoresis includes, for example, one or more syringes or spotters, and can be positioned at the sample injection position by XY control, and using a sample supply unit capable of automatically controlling the sample injection amount. Is supplied to a predetermined sample injection position of each migration channel.
[0176]
This sample is supplied into the opening 42 when the opening 42 is formed. When the first and second electrodes 50 and 52 electrically connected to the migration channel 30 are formed and the opening 42 does not exist, the first electrode 50 and the second electrode 52 Is supplied to the exposed region of the migration channel 30 located at the position (1).
[0177]
Next, an outline of an electrophoresis process using the electrophoresis apparatus of the present invention will be described.
[0178]
<First method>
The first method is an electrophoresis process applied to an electrophoresis device in which the first and second electrodes 50 and 52 are formed.
[0179]
First, the electrophoresis apparatus 10 into which the buffer for electrophoresis has been injected as described above is prepared.
[0180]
Next, on a predetermined sample injection position of the electrophoresis apparatus 10, for example, on the electrophoresis channel 30 located between the first and second electrodes 50 and 52, in particular, to increase the migration length, the first or the first The sample is injected either on the edge 32a or 32b near the second electrode or onto the electrophoresis channel 30 exposed in the opening 42 (see FIG. 8). Here, an example in which a sample is injected onto the migration channel 30 exposed at the edge 32a near the first electrode 50 to perform electrophoresis will be described.
[0181]
Next, in this example, a negative (−) pole is connected to the first electrode 50 near the starting point of electrophoresis, and a positive (+) pole is connected to the second electrode 52, and an electric field is applied to the migration channel 30. Perform electrophoresis.
[0182]
<Second method>
The second method is an example in which electrophoresis is performed while supplying (discharging) the electrophoresis buffer from both sides of the first edge 32a and the second edge 32b of the migration channel 30. Specifically, the first and second edge portions 32a and 32b are immersed in separate containers each containing a buffer for electrophoresis.
[0183]
Here, the electrophoresis buffer applied to the first method and the second method described above will be described. As an electrophoresis buffer, for example, a buffer having a composition well known to those skilled in the art according to publications and standard academic books described above can be used. For example, a buffer solution containing 20% by weight of acetonitrile, 50 mM (M: Mol / L) SDS and 50 mM boric acid can be mentioned.
[0184]
When the first and second electrodes 50 and 52 electrically connected to the migration channel 30 are formed, the electrophoresis is performed so that these electrodes do not contact the migration buffer. In addition, as described above, one end of the fibrous fiber is connected to the first edge 32a or the second edge 32b, and the other ends are placed in separate containers each containing a buffer for electrophoresis. It may be soaked.
[0185]
At this time, the buffer solution contained in the two different containers may be a buffer solution suitable for desired electrophoresis. For example, they may be buffers having the same composition as each other, or buffers having different compositions. Further, the composition of the electrophoresis buffer solution previously injected into the electrophoresis channel 30 may be different from the composition of these buffer solutions.
[0186]
Next, an electric field is applied to the electrophoresis channel 30. When the first edge portion 32a or the first edge portion 32a is immersed in the electrophoresis buffer in a different container, a voltage can be directly applied to the electrophoresis buffer. That is, the positive (+) electrode is brought into contact with the electrophoresis buffer in one container, and the negative (−) electrode is brought into contact with the electrophoresis buffer in the other container. At the same time, an electric field is applied.
[0187]
When the first and second electrodes 50 and 52 electrically connected to each of the migration channels 30 are formed, one of the electrodes has a positive (+) polarity and the other has a negative (+) polarity. -) Electrophoresis can also be performed by applying an electric field individually for each migration channel, for example, as a pole.
[0188]
After the electrophoresis is completed, the selected staining reagent is irradiated with specific excitation light, thereby detecting the electrophoresis pattern as fluorescence emission and analyzing it.
[0189]
(Sixth embodiment)
A configuration of an electrophoresis apparatus according to a sixth embodiment of the present invention will be described with reference to FIGS. The components already described in the first to fifth embodiments are denoted by the same reference numerals, and detailed description thereof will be omitted. In addition, the applied materials, the conditions for each step, and the like can be applied in principle to the examples described in the first to fifth embodiments, and thus detailed descriptions thereof may be omitted.
[0190]
FIG. 14A is a schematic plan view seen from the top for explaining the configuration of the electrophoresis apparatus according to the sixth embodiment, and FIG. 14B is a cross-sectional view taken along the line II of FIG. FIG. 14C is a schematic cross-sectional view showing a vertical cross section cut along a broken line, and FIG. 14C is a schematic cross-sectional view showing a cross section cut along a II-II ′ broken line in FIG. 14A.
[0191]
The electrophoresis apparatus 10 according to the sixth embodiment has a configuration suitable for use in so-called two-dimensional electrophoresis, particularly in second-dimension electrophoresis using a molecular weight as an index. That is, the electrophoresis channel 30 of the electrophoresis apparatus 10 according to the sixth embodiment stores a sample to be subjected to the first dimension electrophoresis, that is, the second dimension electrophoresis after the isoelectric focusing. A part of the electrophoresis channel 30 that is in contact is formed so as to expand so that the contact surface with the sample becomes larger.
[0192]
Further, the electrophoresis apparatus 10 according to the sixth embodiment has a structure in which the first channel is provided on a region where a sample can be supplied to each migration channel in accordance with the migration channel 30 including the region whose area is expanded as described above. It has a fixing frame 90 for fixing the sample after the electrophoresis of the dimension.
[0193]
The configuration of the migration channel 30 and the configuration in which the fixed frame 90 is provided are applicable to any of the electrophoresis apparatuses described in the above-described first to fifth embodiments, for example, in order to more efficiently inject a sample. can do.
[0194]
Therefore, detailed description of the configuration other than the migration channel 30 and the fixed frame 90 is omitted here.
[0195]
The electrophoresis apparatus 10 includes a substrate 20. A plurality of migration channels 30 are formed on the first main surface 20a of the substrate 20. The linear migration channels 30 are formed parallel to each other and apart from each other along the longitudinal direction of the electrophoresis apparatus 10. In this configuration example, in the electrophoresis channel 30, the region where the sample on which the first-dimensional electrophoresis has been completed is provided is greatly expanded in the vertical direction in the drawing so as to be circular. In this example, the shape is circular. However, the shape is not particularly limited, and may be any suitable shape such as a rectangular shape.
[0196]
The migration channel 30 is formed of a porous body.
[0197]
On the first main surface 20a of the substrate 20 on which the migration channel 30 is formed, a migration channel protective film 40 is formed so that the edge 32 of the migration channel 30 is exposed.
[0198]
In the channel protective film 40, an opening 43 for a fixed frame is formed so as to extend over the plurality of migration channels 30. The opening 43 for the fixing frame is opened in the depth direction from the surface of the channel protective film 40 to the surface of the migration channel 30. At this time, the fixed frame opening 43 is provided so that a fixed frame 90 described later can be fitted therein. The fixed frame opening 43 has the same shape as the contour of the outer shape of the fixed frame 90, is formed to have a size that allows the fixed frame 90 to be fitted without any gap, and is formed so as to straddle each migration channel 30. Good to do.
[0199]
A fixed frame 90 is fitted into the opening 42. The fixing frame 90 is preferably configured to be bonded by an adhesive (not shown) or the like. The fixed frame 90 is provided with a through hole 92. It is preferable that the size of the through hole 92 is such that a sample on which the first dimension electrophoresis has been completed is provided and can be fixed. The fixing frame 90 is preferably made of glass.
[0200]
The opening 43 for the fixing frame can have an appropriate shape and an appropriate size according to the shape and size of the fixing frame 90.
[0201]
In addition, similarly to the above-described embodiment, the opening 43 for the fixed frame is provided with the migration channel 30 between the first electrode 50 and the second electrode 52 of each migration channel 30 in accordance with the fixed frame 90. If it is above, it can be formed at an appropriate position.
[0202]
First and second electrodes 50 and 52 are formed on the exposed edge 32 of the migration channel 30.
[0203]
Further, in the electrophoresis apparatus according to the present embodiment, after the sample on which the first-dimensional electrophoresis has been completed is fixed to the fixing frame 90, the sample is pressed from the upper surface side so that the sample can be easily transferred to the migration channel 30. Such a pressing means may be added. For example, a sample (eg, an immobilized pH gradient gel) on which first-dimensional electrophoresis has been completed is placed in the fixed frame 90, and the sample can be pressed against the migration channel 30 with a uniform force. Means may be used.
[0204]
According to the electrophoresis apparatus of the sixth embodiment, the second-dimensional electrophoresis can be efficiently performed without adjusting a new sample for the second-dimensional electrophoresis of the two-dimensional electrophoresis. can do.
[0205]
The method of manufacturing the electrophoresis apparatus according to the sixth embodiment is the same as the other embodiments except for the configuration of the electrophoresis channel 30 and the configuration in which the fixed frame 90 is provided in the opening 43 for the fixed frame. Detailed description is omitted. Therefore, only the step of forming the migration channel 30 and the step of providing the fixed frame 90 will be described. As described above, the step of forming the migration channel 30 includes (1) the step of forming a groove in advance described in the first embodiment, and (2) the formation on the entire surface of the substrate described in the second embodiment. And a step of patterning the film.
[0206]
The case where the step (1) is applied to the manufacturing method of this embodiment will be schematically described. As described above, the region where the sample on which the first-dimensional electrophoresis has been completed is provided is, for example, circular. In this case, the substrate is etched using the resist layer as a mask so that the shape pattern of the migration channel 30 including the partial region greatly expanded in the vertical direction is circularly exposed on the substrate surface. By this etching, a groove having a desired shape, that is, a linear or curved portion is formed. Then, a porous body is deposited so as to fill the groove to form a migration channel.
[0207]
The case where the step (2) is applied will be schematically described. On the entire surface of the substrate, a porous film constituting the migration channel is formed to have a predetermined thickness of the migration channel set in advance. Next, for example, by applying a conventionally known general condition, by a plasma CVD method using an ethyl silicate (TEOS) gas, a SiO 2 for protecting the film for forming the migration channel is formed. ₂ Is formed on the entire surface.
[0208]
Next, by a photolithography process under the same conditions as those applied in a conventionally known semiconductor device manufacturing process, the region in which the sample on which the first-dimensional electrophoresis has been completed is provided, for example, in a vertical direction so as to be circular. A resist layer is formed in the shape pattern of the migration channel 30 including the greatly expanded portion.
[0209]
Next, using the patterned resist layer as a mask, the exposed channel formation film is etched to form a pattern of the migration channel 30.
[0210]
The migration channel 30 or the film before patterning for forming the migration channel 30 may be formed by any one of the methods (1) to (4) described in the first embodiment.
[0211]
The opening 43 for the fixed frame is formed so that a part of the surface of the migration channel 30 is exposed by, for example, reactive ion etching using oxygen plasma using the resist layer as a mask, as in the above-described embodiment. Can be. The fixing frame 90 having the above-described shape and size is provided in the fixing frame opening 43 by, for example, fitting and bonding.
[0212]
(Seventh embodiment)
Referring to FIG. 15, the configuration of the electrophoresis apparatus according to the seventh embodiment of the present invention will be described. The same components as those described in the first to sixth embodiments are denoted by the same reference numerals, and detailed description thereof will be omitted. The applied materials and the conditions of each step can be applied in principle to the examples described in the first to sixth embodiments, and thus detailed description thereof is omitted.
[0213]
FIG. 15 is a schematic cross-sectional view illustrating a cross section cut across a migration channel for describing a configuration of an electrophoresis apparatus according to a seventh embodiment.
[0214]
The electrophoresis apparatus 10 according to the seventh embodiment has a configuration for analyzing an electrophoresis result from the second main surface 20b side of the substrate 20. That is, the substrate 20 of the electrophoresis apparatus 10 according to the seventh embodiment has at least the electrophoresis channel 30 on the second main surface 20b side such that the electrophoresis result can be analyzed from the second main surface 20b side. The depression 24 is formed so as to be thin over the formed region.
[0215]
The configuration of the seventh embodiment can be applied to any of the electrophoresis devices described in the first to sixth embodiments. Therefore, a detailed description of the configuration other than the substrate 20 is omitted.
[0216]
The electrophoresis apparatus 10 includes a substrate 20. The substrate 20 has a plurality of migration channels 30 formed therein. In the configuration example of this embodiment, since the analysis of the electrophoresis result is performed from the second main surface 20b side, the material of the substrate 20 is preferably a light-transmitting plate. Preferably, for example, quartz glass or non-fluorescent glass is used. In this case, the recess 24 should be formed on the second main surface 20b side so as to be thinner at least over the region where the migration channel 30 is formed until it is about 0.1 mm from the bottom of the migration channel 30. Good.
[0219]
By doing so, for example, a material having low light transmittance, such as silicon nitride, can be used as the material of the channel protective film 40. Expands. Therefore, a cheaper electrophoresis apparatus can be provided.
[0218]
The method of manufacturing the electrophoretic device according to the seventh embodiment is the same as that of the other embodiments except for the configuration in which the depression 24 is provided in the substrate 20, and thus the detailed description is omitted, and the depression 24 is provided. Only the steps will be described.
[0219]
The depression 24 is formed by a suitable process according to the material constituting the substrate 20.
[0220]
For example, the case where the substrate 20 is made of quartz glass will be described. First, a mask is formed on the second main surface 20b by a photolithography method. This mask is formed so that the region where the depression 24 is formed is exposed. Then, by wet etching with hydrofluoric acid under conventionally known reaction conditions, the electrode is formed to a depth that allows analysis after migration, that is, observation of the migration channel 30, from the second main surface 20b side. In this configuration example, the depth of the recess 24 is preferably formed to be preferably about 0.1 mm.
[0221]
This step is preferably performed after the formation of the channel protection film 40, but may be performed before the formation of the migration channel 30 or the channel protection film 40, provided that the configuration of the migration channel 30 is not damaged.
[0222]
The electrophoretic device of the present invention includes the components in each of the embodiments described above, for example, the material of the substrate 20, the shape of the migration channel 30, the material of the channel protective film 40, the opening 42, and the electrodes 50 and 52. The material and the arrangement position can be appropriately selected and combined.
[0223]
In the description of the electrophoresis apparatus 10 of the present invention, the substrate 20 has been described as a plate-like body in which the first main surface 20a and the second main surface 20b have the same shape and the same rectangular shape as each other. Not limited. The shape may be, for example, a circle, an ellipse, or the like as long as the object of the invention is not impaired.
[0224]
In the description of the electrophoresis apparatus 10 of the present invention, the configuration example in which the migration channel 30 extends in the longitudinal direction of the apparatus 10 has been mainly described, but the configuration is not limited thereto. For example, the second electrophoresis channel may be formed in a direction orthogonal to the linear electrophoresis channel 30 extending in the longitudinal direction, for example, so that two-dimensional electrophoresis can be performed.
[0225]
Furthermore, in the description of the electrophoresis apparatus 10 of the present invention, an example in which the migration channel is formed so as to be exposed on the side surface of the substrate has been described. The edge of the electrophoresis channel can be set inside the periphery defining the surface so as not to be exposed on the side surface.
[0226]
In the description of the electrophoresis apparatus 10 of the present invention, an example in which a sample for electrophoresis is a protein has been described. For example, the size of pores may be formed in any suitable size in consideration of the size of a substance to be electrophoresed. For example, it can be subjected to electrophoresis of a biologically-related substance such as a nucleic acid other than a protein, ie, a DNA or RNA fragment.
[0227]
In manufacturing the electrophoresis apparatus of the present invention, the following suitable steps can be performed.
(1) forming a resist layer having a pattern for forming a groove on the first main surface of the substrate;
(2) forming a groove through a resist layer;
(3) a step of applying a sol-like mixed solution on the first main surface via a resist layer so as to fill the grooves;
(4) a step of converting the applied sol-like mixed solution into a migration channel formed of a porous body by heating;
(5) removing the resist layer;
(6) A step of forming a channel protective film on the first main surface such that an edge of the migration channel is exposed.
[0228]
Preferably, the step (2) is performed by reactive ion etching.
[0229]
Also, the step (5) preferably includes a step of flattening the upper surface of the formed migration channel.
[0230]
Further, the step (6) preferably includes a step of forming first and second electrodes at the edge of the migration channel.
[0231]
Furthermore, the step (6) preferably includes a step of forming an opening on the channel protective film.
[0232]
【The invention's effect】
According to the configuration of the electrophoresis apparatus of the present invention, electrophoresis can be performed in a state of being separated for each of a plurality of migration channels, so that a large number of samples can be simultaneously analyzed in a short time and with higher resolution. Can be.
[0233]
In the case where each of the first and second electrodes 50 and 52 is formed in each of the first to n-th migration channels 30-1 to 30-n, the respective migration channels 30 are independent of each other. Since the electrophoresis can be performed by applying an electric field to the device, the electrophoresis apparatus 10 can be operated flexibly. In the case where the opening 42 is formed, more accurate sample supply to the migration channel 30 can be performed.
[0234]
According to the configuration of the electrophoresis apparatus of the fifth embodiment, the length of the migration channel 30 can be made longer, so that the resolution in analysis can be improved.
[0235]
According to the electrophoresis apparatus of the sixth embodiment, the second dimension electrophoresis can be efficiently performed without adjusting a new sample for the second dimension electrophoresis of the two-dimensional electrophoresis. be able to.
[0236]
According to the electrophoresis apparatus of the seventh embodiment, for example, a material having low light transmittance such as silicon nitride can be used as the material of the channel protective film 40, so that the film is excellent in film formability and moisture resistance. The range of choices of materials, such as materials, is expanded. Therefore, a cheaper electrophoresis apparatus can be provided.
[Brief description of the drawings]
FIG. 1A is a schematic plan view of the configuration of an electrophoresis apparatus according to a first embodiment, as viewed from above, and FIG. 1B is a plan view of FIG. 1C is a schematic cross-sectional view showing a cut surface taken along a line II ′ of FIG. 1A, and FIG. 1C is a schematic cross-sectional view showing a cut surface taken along a II-II ′ broken line of FIG. FIG.
FIGS. 2A to 2C are schematic cross-sectional views each showing a cut surface taken along the line II-II ′ of FIG. 1A.
FIGS. 3A and 3B are schematic cross-sectional views showing a cross section taken along the line II-II ′ of FIG. 1A.
4 (A) to 4 (C) are schematic cross-sectional views showing a cross section taken along the line II ′ of FIG. 1 (A).
FIG. 5A is a schematic plan view of the configuration of the electrophoresis apparatus according to the second embodiment as viewed from above, and FIG. 5B is a plan view of FIG. 5C is a schematic cross-sectional view showing a cross-section taken along the line II ′ of FIG. 5, and FIG. 5C is a schematic cross-section showing a cross-section taken along the line II-II ′ of FIG. FIG.
6 (A) to 6 (D) are schematic cross-sectional views each showing a cut surface taken along the line II-II ′ of FIG. 5 (A).
FIGS. 7A and 7B are schematic cross-sectional views each showing a cross section taken along the line II ′ of FIG. 5A.
FIG. 8A is a schematic plan view of the configuration of the electrophoresis apparatus according to the third embodiment as viewed from above, and FIG. 8B is a plan view of FIG. FIG. 8C is a schematic cross-sectional view showing a cut surface taken along the line II ′ of FIG. 8, and FIG. 8C is a schematic cross-sectional view showing a cut surface taken along the line II-II ′ of FIG. FIG.
9 (A) to 9 (C) are schematic cross-sectional views each showing a section taken along the line II-II ′ of FIG. 8 (A).
FIGS. 10A to 10D are schematic cross-sectional views each showing a cross section taken along the line II ′ of FIG. 8A.
FIG. 11A is a schematic plan view illustrating the configuration of an electrophoresis apparatus according to a fourth embodiment, viewed from above, and FIG. 11B is a plan view of FIG. 11C is a schematic cross-sectional view showing a cut surface taken along the line II ′ of FIG. 11, and FIG. 11C is a schematic cross-sectional view showing a cut surface taken along the II-II ′ broken line of FIG. FIG.
12 (A) to 12 (D) are schematic cross-sectional views each showing a cross section taken along the line II ′ of FIG. 11 (A).
FIGS. 13A and 13B are schematic plan views illustrating the configuration of an electrophoresis apparatus according to a fifth embodiment, as viewed from above. FIG. 13A shows a first configuration example. FIG. 13B shows a second configuration example.
FIG. 14A is a schematic plan view illustrating the configuration of an electrophoresis apparatus according to a sixth embodiment, viewed from above, and FIG. 14B is a plan view of FIG. 14C is a schematic cross-sectional view showing a cut surface taken along the line II ′ of FIG. 14, and FIG. 14C is a schematic cross-sectional view showing a cut surface taken along the broken line II-II ′ of FIG. FIG.
FIG. 15 is a schematic cross-sectional view illustrating a cross section cut across an electrophoresis channel for describing a configuration of an electrophoresis apparatus according to a seventh embodiment.
[Explanation of symbols]
10: Electrophoresis device
20: Substrate
20a: first main surface
20b: 2nd main surface
22: groove
24: hollow
30: Electrophoresis channel
30-1: First migration channel
30-2: Second migration channel
30-3: Third electrophoresis channel
30-n: n-th migration channel
30 ': film for forming a channel
30a: straight section
30b, 30c: curved part
32: Edge
32a: 1st edge part
32b: second edge
34: protective film
40: Electrophoresis channel protective film
42: Opening
43: Opening for fixed frame
50: first electrode
52: second electrode
60: First electrophoresis buffer supply (discharge) means
62: Second electrophoresis buffer supply (discharge) means
70: resist layer
80: protective film for exposing migration channel
90: Fixed frame
92: Through hole

Claims (13)

A first main surface, a substrate having a second main surface opposed to the first main surface, a plurality of grooves formed in the first main surface, and formed to fill the grooves. , A first and second edge formed at both ends of the migration channel, and the first and second edges are exposed. And an electrophoresis channel protective film formed as described above. A first main surface, a substrate having a second main surface opposed to the first main surface, a plurality of migration channels formed of a porous body formed on the first main surface, A first edge portion and a second edge portion formed at both ends of the migration channel, and a migration channel protection film formed such that the first and second edge portions are exposed. An electrophoresis apparatus, comprising: A first main surface, a substrate having a second main surface opposed to the first main surface, a plurality of grooves formed in the first main surface, and electrophoresis formed in the grooves Channel, first and second edges formed at both ends of the migration channel, and migration channel formed so that the first and second edges are exposed A protection film, an opening for a fixed frame formed on the migration channel protection film so as to extend over the plurality of migration channels, and a first dimension provided in the opening for the fixed frame. A fixing frame having a through hole for fixing the sample after electrophoresis,
Electrophoresis, wherein a part of the exposed electrophoresis channel, with which the sample subjected to the first dimension electrophoresis comes into contact, is expanded so that a contact surface with the sample becomes larger. apparatus. The electrophoresis apparatus according to any one of claims 1 to 3, wherein each of the plurality of electrophoresis channels has a linear shape and is formed to be spaced apart in parallel with each other. The electrophoresis device according to any one of claims 1 to 4, wherein the migration channel is formed in a wave shape combining a plurality of curved portions. A plurality of the electrophoresis channels, a linear portion including a plurality of linear electrophoresis channels formed in parallel to each other, and a predetermined number of the plurality of linear electrophoresis channels as a set, adjacent to each set The electrophoretic device according to any one of claims 1 to 5, wherein the linear electrophoresis channel has a folded structure in which ends of the linear electrophoresis channel are connected by a curved portion. The electrophoresis apparatus according to claim 1, wherein the migration channel is formed of porous silicon. The substrate is formed of a light-transmitting material, and the depression for analyzing the migration channel formed on the substrate from the second main surface side is formed with the migration channel of the substrate. The electrophoresis apparatus according to claim 1, wherein the electrophoresis apparatus is formed in a region where the electrophoresis is performed. The electrophoresis apparatus according to claim 8, wherein the substrate is made of quartz glass. The electrophoresis channel protective film located between the first and second edge portions has a plurality of openings for exposing a region where a sample can be supplied to each of the electrophoresis channels. The electrophoresis apparatus according to claim 1. The method of claim 1, further comprising a first electrode electrically connected to a first edge of the migration channel, and a second electrode electrically connected to a second edge of the migration channel. Item 11. The electrophoresis apparatus according to any one of Items 1 to 10. A plurality of first electrodes formed on the first edge portion electrically connected to each of the plurality of electrophoresis channels, and a plurality of second electrodes formed on the second edge portion; The electrophoresis apparatus according to claim 11, further comprising an electrode. The electrophoresis buffer according to claim 1, further comprising an electrophoresis buffer supply means connected to the electrophoresis channel such that an electrophoresis buffer can be supplied to a first edge portion and a second edge portion of the electrophoresis channel. 13. The electrophoresis apparatus according to any one of 12 above.

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