JP5504131B2 - Pretreatment equipment - Google Patents

Description

  The present invention relates to a pretreatment instrument, and more particularly, to a pretreatment instrument used for processing a sample such as electrophoresis in which pretreatment materials such as various reagents are sealed.

  After the completion of the Human Genome Project, proteome research has been actively conducted. This “proteome” means the total of proteins produced by translation in specific cells, organs, and organs, and examples of research on the proteome include protein profiling.

  As one of the techniques for profiling proteins, two-dimensional electrophoresis of proteins is known. This two-dimensional electrophoresis is composed of a total of two electrophoresis steps: isoelectric focusing that separates proteins depending on charge and slab gel electrophoresis that separates proteins depending on molecular weight (especially SDS-PAGE). (For example, refer to Patent Document 1).

  An example of a sample separation apparatus that performs the two-dimensional electrophoresis is disclosed in Patent Document 2. The sample separation device includes a first chip that is separated by isoelectric focusing and a second chip that performs slab gel electrophoresis.

The first chip is composed of a support plate and a first gel (first-dimensional gel) supported on the support plate. In this first chip, a sample (sample) is introduced into the first gel, and after passing through the step of swelling the first gel, a voltage is applied to the first gel to separate the sample in the first direction ( First dimension separation). Then, the first chip is transferred to the second chip after being subjected to the process of staining the separated sample and the process of equilibrating to the environment of the second chip.
The second chip is configured by storing a second gel (second-dimensional gel) in the substrate, and the first gel of the first chip is connected to the second gel. In this state, by applying a voltage to the second gel, the sample is separated in a second direction different from the first direction (two-dimensional separation).

  Patent Document 2 describes that by automating each process of two-dimensional electrophoresis, the convenience of two-dimensional electrophoresis is improved, the dependency on the operator is reduced, and the reproducibility of the results is improved. .

Japanese Patent Laid-Open No. 11-30605 JP 2007-64848 A

  By the way, the process of introducing the sample into the first dimension gel performed before the first dimension separation, the reaction process with the necessary reagents performed after the first dimension separation and before the second dimension separation, etc. Usually, it is carried out using a pretreatment instrument having a plurality of groove structures. In each of the plurality of groove structures, pretreatment materials such as various reagents and cleaning agents are singly or prepared, and the first gel held by the first chip is sequentially brought into contact with the pretreatment material in each groove. Each process is advanced by this.

In such a pretreatment instrument, there is an attempt to improve convenience by filling the groove with a pretreatment material in advance and sealing with a sealing material until use.
However, when the pretreatment material is liquid, the inventors have peeled off the sealant during use, so that the pretreatment material adhering to the sealant leaks to the outside of the pretreatment instrument or in other grooves. It has been found by examination that there is a possibility that it may interfere with the experiment.

  The present invention has been made in view of the above circumstances, and provides a pretreatment instrument that seals the inside of the groove suitably before use and can be easily opened during use, and does not easily leak or mix pretreatment materials. The purpose is to do.

  A first aspect of the present invention is a pretreatment instrument for pretreating a sample, a base material having a plurality of grooves opened on an upper surface, and a pretreatment material disposed in at least one of the plurality of grooves And a sealing material for sealing at least two or more of the plurality of grooves, the sealing material having a surface sheet coated with a release agent on the first surface, and a heat on one surface. A bonding layer having a fusible layer and bonding the heat-fusible layer to the base material and the first surface of the top sheet and the other surface of the film material And a first half-cut portion formed from the top sheet side to at least a depth penetrating the top sheet in the thickness direction, and at least the joining layer from the one surface side of the film material in the thickness direction. A second half-cut portion formed to a depth penetrating through The cut portion has two half cuts extending at a narrower interval than the width of the predetermined groove in which the pretreatment material is disposed among the plurality of grooves, and the first half cut portion is When the sealing material is viewed in the thickness direction, the sealing material has a pair of half cuts extending so as to sandwich the two half cuts. In the plan view, the base material is fused to the upper surface of the base material so as to be positioned within the width of the predetermined groove.

  A second aspect of the present invention is a pretreatment instrument for pretreating a sample, a base material having a groove opened on an upper surface, a pretreatment material disposed in the groove, and sealing the groove. A sealing material, the sealing material has a surface sheet, a film material on which one surface has a heat-fusible layer, and the heat-fusible layer is fused to the substrate; The surface sheet and a bonding layer for releasably bonding the other surface of the film material, the film material has a through-hole having a narrower width than the groove, and the sealing material The through hole is fused to the base material so as to be positioned within the width of the groove in a plan view of the pretreatment instrument.

  A third aspect of the present invention is a pretreatment instrument for pretreating a sample, a base material having a groove opened on an upper surface, a pretreatment material disposed in the groove, and sealing the groove. A sealing material, the sealing material has a surface sheet, a film material on which one surface has a heat-fusible layer, and the heat-fusible layer is fused to the substrate; The surface sheet and a bonding layer that releasably bonds the other surface of the film material, and the film material has a depth that penetrates at least the film material in the thickness direction from the one surface side. A half-cut portion including two half-cuts that are formed at a distance narrower than the width of the groove, and the sealing material has the half-cut portion in a plan view of the pretreatment instrument. And being fused to the base material so as to be positioned within the width of the groove

The heat-fusible layer may be formed only on a portion to be fused to the base material on the one surface of the film material.
Moreover, the site | part in which the said heat-fusible layer is not formed in the said one surface of the said film material may be hydrophobic.

  According to the pretreatment instrument of the present invention, the inside of the groove is preferably sealed before use, and can be easily opened at the time of use, so that the pretreatment instrument is less prone to leakage and mixing of the pretreatment material.

It is a top view which shows the base material in the pretreatment instrument of 1st Embodiment of this invention. It is sectional drawing in the aa line of FIG. It is a figure which shows the layer structure of the sealing material in the pre-processing instrument. It is sectional drawing which shows the completed said pretreatment instrument.

Hereinafter, a pretreatment instrument according to a first embodiment of the present invention will be described with reference to FIGS. 1 to 4.
The pretreatment instrument of this embodiment is an instrument having a function of performing various pretreatment steps before and after sample introduction with respect to the first chip used for two-dimensional electrophoresis, and a base material on which a plurality of grooves are formed; And a sealing material for sealing the groove of the base material.

FIG. 1 is a plan view showing a base material 10 of the pretreatment instrument 1 of this embodiment, and FIG. 2 is a cross-sectional view taken along the line aa in FIG.
As shown in FIGS. 1 and 2, the base material 10 is formed with a certain thickness, and has a plurality of grooves 11 that open to the upper surface 10A and extend in the same direction. The material of the substrate 10 is not particularly limited as long as the sealing material described later can be heat-sealed, and various resins may be appropriately selected in consideration of chemical resistance and the like.

  There are no particular restrictions on the number, width, length, installation interval, etc. of the grooves 11, depending on the dimensions of the first chip to be inserted, the number and content of pretreatment to be performed, the required amount of pretreatment material, etc. It may be set appropriately. Therefore, for example, as in the groove 11A shown in FIG.

As shown in FIG. 2, a plurality of types of pretreatment materials such as chemicals, reagents, and solvents used for various pretreatments are arranged in the groove 11.
Examples of the pretreatment in this embodiment include introduction of a sample, swelling of the first gel, grounding of the first gel, washing, replacement with a buffer, intermediate staining, protein SDS treatment, washing of the electrodes, etc. Drugs necessary for the treatment are appropriately selected and arranged. There is no limitation on the form of the pretreatment material, and it may be arranged in various forms such as a solid such as a gel or a powder in addition to a liquid, and a plurality of pretreatment materials may be mixed as needed. It may be arranged in the groove 11.

FIG. 2 shows an example in which different pretreatment materials 12A to 12H are arranged in the eight grooves 11 including the grooves 11A as an example, but it is necessary to arrange the pretreatment materials in all the grooves. Alternatively, the position of the groove in which the pretreatment material is disposed may be appropriately set in consideration of the order of each treatment.
For example, when a process using another reagent or the like prepared at the time of use is performed between the process using the pretreatment material 12A and the process using the pretreatment material 12B, the pretreatment material 12A is disposed. An empty groove in which the pretreatment material is not disposed may be secured between the groove and the groove in which the pretreatment material 12B is disposed.

  FIG. 3 is a diagram showing a layer structure of the sealing material 20 that seals the groove 11. The sealing material 20 includes a surface sheet 21, a film material 22 bonded below the surface sheet 21, a bonding layer 23 that bonds the surface sheet 21 and the film material, and the film material 22. And a heat-fusible layer 24 formed on the lower surface (one surface).

  As the material of the topsheet 21, a polyolefin film, a PET film, paper, or the like can be used. In the surface sheet 21, the lower surface (first surface) to which the film material 22 is bonded is provided with a release agent 25 made of a mixture of a surfactant such as lecithin, a nitrified cotton-based adhesive, wax, silicon or the like. Is applied.

  As the film material 22, a material made of a material such as a heat-resistant resin that can be heat-sealed can be appropriately selected and used in consideration of gas barrier properties and the like.

  As the material of the bonding layer 23, a material having excellent thermal adhesiveness with the lower surface of the surface sheet 21 to which the release agent 25 is applied, specifically, polyethylene, EMA (ethylene-methacrylic acid copolymer), or the like is used. can do.

The material of the heat-fusible layer 24 does not exhibit adhesiveness at room temperature, and exhibits adhesiveness when heated to a predetermined temperature and is fused to the upper surface 10A of the substrate 10. After the fusion, when it is cooled to room temperature, the heat-fusible layer 24 loses its adhesiveness again and does not affect the pretreatment material disposed in the groove 11.
The heat-fusible layer 24 may be evenly laminated on the lower surface of the film material 22 by a technique such as a co-extrusion method, or may be printed only on a part of the lower surface, for example, a portion to be fused to the upper surface 10A of the base material 10. Or the like.
Examples of the heat-fusible material suitable for such a forming method include various resins such as polyolefin-based, polypropylene-based barrier coextrusion systems, and polyolefin-based polymer alloys modified with vinyl polymers.

The heat-fusible layer 24 may be provided by a coating method other than the above-described method. In this case, a heat seal lacquer composition can be suitably used. The material of the heat seal lacquer composition is not particularly limited, but ethylene-vinyl acetate copolymer (EVA), polyvinylidene chloride, chlorinated polyolefin, polyamide, acrylic resin, saturated polyester, vinyl chloride-vinyl acetate copolymer, polyolefin As the base resin, a thermoplastic resin such as a single resin or a mixture thereof can be used.
The heat seal lacquer composition may be constituted by appropriately blending these materials with additives such as wax, tackifier, plasticizer, antiblocking agent, slip agent, stabilizer, and curing agent.

  As shown in FIG. 4, the sealing material 20 is half-cut so that only a part of the plurality of grooves 11 can be opened. Two half cuts 26 formed from the heat-fusible layer 24 side to at least a depth penetrating the bonding layer 23 are formed so as to extend substantially parallel to each other in the longitudinal direction of the groove 11, and the interval is the width of the groove to be opened. Set smaller than the dimension. Thereby, the second half-cut portion 32 by the half-cut 26 is formed in the sealing material 20. The end portions of the two half cuts 26 may be connected, and the second half cut portion 32 may be formed in a rectangular shape in plan view.

  On the other hand, the pair of half cuts 27 formed from the surface sheet 21 side to at least a depth penetrating the surface sheet 21 extend substantially parallel to each other over the longitudinal direction of the groove 11 and are second in the plan view of the pretreatment instrument 1. The half-cut portion 32 is formed so as to be positioned therebetween. Thereby, the first half cut portion 31 by the half cut 27 is formed in the sealing material 20. The first half-cut portion 31 may also be formed in a rectangular shape in plan view. In this case, the second half-cut portion 32 is formed in the rectangle formed by the first half-cut portion 31 in the plan view of the pretreatment instrument. The rectangle to be formed should be positioned.

The sealing material 20 configured as described above was cut into an appropriate size in accordance with the base material 10 on which the pretreatment material was disposed, and the heat-fusible layer 24 was brought into contact with the upper surface 10A of the base material 10. When the heat-fusible layer 24 is heated by a known thermocompression bonding means in the state, the heat-fusible layer 24 and the upper surface 10A of the base material 10 are integrally fused, and the pretreatment instrument 1 shown in FIG. 4 is completed. To do.
In the present embodiment, the first half-cut portion 31 and the second half-cut portion 32 can peel only a groove 11 containing the pretreatment material 12 </ b> C by peeling a part of the sealing material 20. The pretreatment instrument 1 is configured. Furthermore, perforations 28 that penetrate the sealing material 20 in the thickness direction and intermittently extend in the longitudinal direction of the groove are formed in portions of the sealing material 20 located on both sides in the width direction of the groove. Has been. Thereby, a part of sealing material 20 can be removed and another groove | channel can be opened with the said groove | channel sealed.

The sealing material 20 heat-sealed to the pretreatment instrument 1 has an easy peel property that allows the user to easily peel it off by hand during use. Therefore, in the distribution stage before use, etc., the groove 11 in which the pretreatment material is disposed is sealed to suitably prevent the entry of foreign matter from the outside, evaporation of the pretreatment material, scattering, leakage, alteration, etc. At the time of use, it is easily peeled off and can be suitably used for pretreatment of the first chip.
Further, since the interval between the half cuts 26 in the second half cut portion 32 for opening the predetermined groove is set smaller than the width dimension of the target groove, the sealing material is also provided after the groove is opened. A part of 20 covers the upper part of the groove. As a result, the pretreatment material 12C in the groove can be pretreated efficiently by suitably suppressing such a situation that it is unintentionally scattered during the operation or volatilizes during the experiment.

  The peeling mode of the sealing material 20 is divided into a plurality of types such as cohesive peeling, delamination, and interface peeling depending on the material of the heat-fusible layer 24. It can be used. Among these, in the case of interfacial peeling, since the heat-fusible layer 24 does not remain on the substrate 10 side after the sealing material 20 is peeled off, fine fragments or the like of the heat-fusible layer 24 are mixed in the groove 11. The occurrence of such a situation can be further reduced, which is preferable.

The pretreatment instrument 1 of the present embodiment configured as described above will be described in more detail using examples. In addition, the following examples are for confirming the sealing performance and easy peel property of the sealing material, and no pretreatment material was disposed in the groove.
(Example)
The base material 10 was produced by injection molding using an acrylic resin. The substrate 10 had dimensions of 70 millimeters (mm) in width, 70 mm in length, and 10 mm in thickness. Six grooves 11 having a width of 5 mm, a length of 40 mm, and a depth of 5 mm were formed at intervals of 5 mm.

  The sealing material 20 was produced as follows. First, a film material (manufactured by J-Film Co., Ltd., product) formed by extrusion lamination of a polyethylene-based heat-fusible layer 24 on the PE-side surface of a film comprising two layers of polyethylene terephthalate (PET) and polyethylene (PE) The name SMXZH44) was used as the film material 22.

  Next, the surface sheet 21 is prepared by applying a release agent 25 formed by mixing silicon into a surfactant on the back side of the PET film, and polyethylene having excellent thermal adhesiveness with the application surface of the release agent 25 The sealing material 20 was produced by bonding the film material 22 and the surface sheet 21 by the extrusion laminating method using the bonding layer 23 made of the above.

  Further, two perforations 28 penetrating in the thickness direction at intervals of 7 mm are formed in the sealing material 20, and a pair of depths penetrating the top sheet 21 from the top sheet 21 side in a region between the perforations 28. The first half-cut portion 31 is formed, the first half-cut portion 31 is provided, and the two half-cuts 26 penetrating from the heat-fusible layer 24 side to the release agent 25 are formed to form the second half-cut portion. 32 was provided. The second half-cut part 32 is formed so as to form a rectangle with a width of 3.5 mm and a length of 35 mm, and the first half-cut part 31 forms a rectangle with a width of 5.5 mm and a length of 40 mm, And when the sealing material 20 was seen in the thickness direction, it formed so that the rectangle which the 2nd half cut part 32 forms might be surrounded.

  One of the grooves 11 formed in the base material 10 was filled with water, the heat-fusible layer 24 of the sealing material 20 was directed to the upper surface 10A of the base material 10, and the second half-cut portion 32 was formed. The region was positioned so as to cover the groove 11 filled with water, and pressure heat fusion was performed under the condition of 150 ° C. for 1 second. As a result, the groove 11 was suitably sealed by the sealing material 20, and the pretreatment instrument 1 was completed.

In the completed pretreatment instrument 1, the portion where the first half-cut portion 31 and the second half-cut portion 32 were formed was easily peeled off to open the groove 11 filled with water. When part of the sealing material 20 was peeled off, the water filled in the groove 11 was not scattered.
Moreover, the other groove | channel 11 was able to be opened without a problem by peeling a part of sealing material 20 divided by the perforation 28. The sealing material 20 had easy peel properties, and the user could easily peel it off by hand.

As described above, according to the pretreatment instrument 1 of the present embodiment, the groove 11 is sealed by the sealing material 20 in a state where the necessary pretreatment material is disposed in the groove 11 formed in the base material 10. It has stopped. Therefore, it can be stored without any problem until the time of use, and at the time of use, the sealing material 20 can be easily peeled off from the base material 10 and used for pretreatment, thereby greatly improving the convenience for the user. Can do.
Moreover, it formed in the area | region which covers at least 1 groove | channel among the sealing materials 20 from the 1st half cut part 31 which has the half cut 27 formed from the surface sheet 21 side, and the heat-fusible layer 24 side. Since the second half-cut portion 32 having the half-cut 26 is formed, only the groove covered with the region where the half-cut portion is formed can be opened to perform the pretreatment operation. Furthermore, as described above, even after the groove is opened, it is possible to suitably suppress the scattering, volatilization, leakage, etc. of the pretreatment material filled in the groove.

  Further, the sealing material 20 includes a heat-fusible layer 24 on one surface, and the sealing material 20 seals the groove 11 when the heat-fusible layer 24 is heated. The groove 11 can be sealed without using an adhesive or the like. As a result, it is possible to suitably prevent alteration or the like due to the pretreatment material previously enclosed in the groove 11 adhering to the adhesive or the like. Further, since the sealing material 20 has an easy peel property, and the adhesive material does not remain at the pasting site of the base material when it is peeled off, a pretreatment material such as a rare reagent during the pretreatment is used as the adhesive material. Risk of sticking to can be greatly reduced.

  Although one embodiment of the present invention has been described above, the technical scope of the present invention is not limited to the above-described embodiment, and various modifications are made to each component without departing from the spirit of the present invention. Or can be deleted.

First, the structure of the sealing material in the pretreatment instrument of the present invention can have various configurations other than the above.
For example, forming a through-hole narrower than the groove where the pretreatment material is arranged in the film material, and joining the surface sheet and the film material in a peelable manner via a peelable adhesive layer (joining layer) or the like A sealing material may be constituted. In this case, when the sealing material is fused to the substrate so that the through hole is positioned within the width of the groove in a plan view of the pretreatment device, the through sheet is exposed by peeling off the surface sheet, and the groove is opened. Therefore, the same effect as the pretreatment instrument of the above embodiment can be obtained.

Moreover, as another example, after joining a surface sheet and a film material so that peeling is possible through the peelable adhesion layer etc., the half cut part of the shape similar to the above-mentioned 2nd half cut part is at least a film. The sealing material may be configured by forming the material to a depth penetrating in the thickness direction. Also in this case, when the sealing material is fused to the base material so that the half-cut portion is located within the width of the groove in a plan view of the pretreatment instrument, the top sheet is peeled off, thereby removing the half of the sealing material. Only the groove covered with the region where the cut portion is formed can be opened, and the same effect as the pretreatment instrument of the above embodiment can be obtained.
Thus, in the sealing material in this invention, the surface sheet and the film material should just be joined so that peeling is possible, and the mold release agent does not need to be apply | coated.

Moreover, in the above-described embodiment, an example in which all the grooves are sealed using one sealing material having a size covering the entire upper surface of the base material has been described. A sealing material cut to a size covering at least one groove may be fused to the upper surface of the base material, and at least one predetermined groove among the sealed grooves may be configured to be independently openable. . Moreover, you may seal all the groove | channels of a base material using several such sealing materials.
Moreover, you may form the site | part used as a handle for making it easy to hold | grip in the 1st half cut part peeled when opening a predetermined groove | channel.
Furthermore, you may comprise the group of a 1st half cut part and a 2nd half cut part, and may comprise the pretreatment instrument of this invention so that a several groove | channel can be opened independently.

  Moreover, when forming a heat-fusible layer only in the site | part melt | fused to a base material, resin may be exposed about the remaining site | parts, or a further process may be given, and hydrophobicity may be improved. This is preferable because the liquid pretreatment material is less likely to adhere to the sealing material.

DESCRIPTION OF SYMBOLS 1 Pretreatment instrument 10 Base material 11, 11A, Grooves 12A, 12B, 12C, 12D, 12E, 12F, 12G, 12H, Pretreatment material 20 Sealing material 21 Surface sheet 22 Film material 23 Bonding layer 24 Heat-fusible layer 25 Release agent 26 Half cut 27 Half cut 31 First half cut part 32 Second half cut part

Claims (5)

A pretreatment instrument for pretreatment of a sample,
A base material having a plurality of grooves opened on the upper surface;
A pretreatment material disposed in at least one of the plurality of grooves;
A sealing material for sealing at least two of the plurality of grooves;
With
The sealing material is
A top sheet with a release agent applied to the first surface;
A film material having a heat-fusible layer on one side, and the heat-fusible layer being fused to the substrate;
A bonding layer for bonding the first surface of the top sheet and the other surface of the film material;
A first half-cut portion formed from the top sheet side to at least a depth penetrating the top sheet in the thickness direction;
A second half-cut portion formed from the one surface side of the film material to a depth penetrating at least the joining layer in the thickness direction;
The second half-cut portion has two half-cuts extending at a narrower interval than the width of a predetermined groove in which the pretreatment material is disposed among the plurality of grooves,
The first half-cut portion has a pair of half-cuts extending so as to sandwich the two half-cuts when the sealing material is viewed in the thickness direction,
The sealant is fused to the base material so that the two half cuts are positioned within the width of the predetermined groove in a plan view of the pretreatment instrument. Processing equipment. A pretreatment instrument for pretreatment of a sample,
A substrate having a groove opening on the upper surface;
A pretreatment material disposed in the groove;
A sealing material for sealing the groove;
With
The sealing material is
A surface sheet;
A film material having a heat-fusible layer on one side, and the heat-fusible layer being fused to the substrate;
A bonding layer for releasably bonding the top sheet and the other surface of the film material;
The film material has a through hole having a narrower width than the groove,
The pretreatment instrument, wherein the sealing material is fused to the base material so that the through hole is positioned within the width of the groove in a plan view of the pretreatment instrument. A pretreatment instrument for pretreatment of a sample,
A substrate having a groove opening on the upper surface;
A pretreatment material disposed in the groove;
A sealing material for sealing the groove;
With
The sealing material is
A surface sheet;
A film material having a heat-fusible layer on one side, and the heat-fusible layer being fused to the substrate;
A bonding layer for releasably bonding the top sheet and the other surface of the film material;
The film material is formed from the one surface side to at least a depth penetrating the film material in the thickness direction, and includes a half cut portion including two half cuts extending at a distance narrower than the width of the groove. Have
The pretreatment instrument, wherein the sealing material is fused to the base material so that the half-cut portion is positioned within the width of the groove in a plan view of the pretreatment instrument.   The said heat-fusible layer is formed only in the site | part fuse | fused with the said base material in the said one surface of the said film material, The any one of Claim 1 to 3 characterized by the above-mentioned. Pre-processing equipment.   The pretreatment instrument according to claim 4, wherein a portion where the heat-fusible layer is not formed is hydrophobic on the one surface of the film material.

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