JP5767839B2 - 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.

For this possibility, the sealing material is configured to include a surface sheet coated with a release agent, a film material fused to the base material, and a bonding layer that bonds the surface sheet and the film material, It has a first half-cut portion formed from the top sheet side and a second half-cut portion formed from the film material side, and the second half-cut portion is a groove in which the pretreatment material is disposed. By having a structure having two half cuts extending at an interval narrower than the width, a part of the groove part is removed while opening the groove filled with the pretreatment material by peeling off the release agent part from the surface sheet side. There is an attempt to prevent leakage of the pretreatment material by leaving a part of the sealing material in the covering state.
However, this sealing material has a complicated structure and takes a little time to manufacture.

  The present invention has been made in view of the above circumstances, and provides a pretreatment instrument that is preferably sealed in the groove before use and can be easily opened during use and is unlikely to leak or mix pretreatment materials. The purpose is to do.

The pretreatment instrument of the present invention is a pretreatment instrument for pretreating a sample, and is an instrument having a function of performing various pretreatment steps before and after sample introduction,
A base material having a plurality of grooves formed in a certain thickness and opening in the upper surface and extending in the same direction;
A pretreatment material disposed in at least one of the grooves and used for the pretreatment;
A sealing material which is formed including a film material having a heat-fusible layer made of a heat-fusible material on one surface, and seals the groove in which the pretreatment material is disposed;
With
At least one of the grooves in which the pretreatment material is disposed has a wall portion protruding inward in the width direction from both sides in the width direction of the groove, and the width of the opening is formed narrower than the groove width,
The sealing material seals the groove in which the pretreatment material is disposed by heat-sealing the heat-fusible layer to the upper surface of the base material.
The sealing material has two cut portions extending in parallel with an interval larger than the width of the opening, and the cut portion is parallel to the groove in which the pretreatment material is disposed and has an opening in a plan view. It is heat-sealed to the base material so as to sandwich it,
Only the area | region between the said cut parts among the said sealing materials can be peeled from the said base material, and the said subject was solved by the peeling aspect of the said sealing material being the interface peeling.
The heat-fusible layer may be formed only at a site that contacts the upper surface of the base material.
Furthermore, the pretreatment instrument of the present invention may have a function of performing various pretreatment steps before and after the sample introduction with respect to the first chip used for two-dimensional electrophoresis.
The present invention is a pretreatment instrument for pretreatment of a sample, a base material having a plurality of grooves opened on an upper surface, a pretreatment material disposed in at least one of the grooves and used for the pretreatment, A sealing material for sealing the groove in which the pretreatment material is disposed, the film including a film material having a heat-fusible layer made of a heat-fusible material on one surface, and the pretreatment At least one of the grooves in which the material is disposed is formed such that the width of the opening is narrower than the width of the groove, and the sealing material has the heat-fusible layer thermally bonded to the upper surface of the substrate. the pretreatment material by being is Ru can seals the arranged the groove.

The sealing material has two cut portions extending in parallel with an interval larger than the width of the opening, and the cut portion is parallel to the groove in which the pretreatment material is disposed and has an opening in a plan view. It may be heat-sealed to the base material so as to be sandwiched, and only the region between the pair of cut portions of the sealing material may be peelable from the base material.
The sealing material WHEREIN: The area | region between the said cut parts may have a holding part for a user to hold | grip at the edge part of a longitudinal direction.

  According to the pretreatment instrument of the present invention, it is possible to realize a configuration in which the inside of the groove is suitably sealed before use and can be easily opened during use, and the pretreatment material is unlikely to leak or mix.

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 an example of the manufacture procedure of the base material. It is a figure which shows an example of the manufacture procedure of the base material. 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. It is sectional drawing which shows the modification of the same pretreatment instrument. It is a top view of the modification of the pretreatment instrument which provided the grasping part. (A) is a top view of the modification of the pre-processing instrument which provided the holding part, (b) is a side view of the pre-processing instrument. It is a top view of the modification of the pretreatment instrument which provided the grasping part. It is a top view of the modification of the pretreatment instrument which provided the grasping part.

Hereinafter, a pretreatment instrument according to an embodiment of the present invention will be described with reference to FIGS. 1 to 7.
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, like the groove 11A shown in FIG. 1, a part of the grooves may be formed with a different width from the other grooves.

In each groove 11, the opening 11 a has wall portions 13 that protrude inward in the width direction from both sides in the width direction of the groove. For this reason, in each groove 11, the width dimension of the opening formed in the opening 11 a is set to be smaller than the width dimension of other portions of the groove 11.
The dimension of the wall 13 that defines the width of the opening may be appropriately set according to the dimension of the first chip to be inserted. Moreover, you may differ for every groove | channel.

  For example, as shown in FIG. 3, the base material 10 on which the groove 11 having the wall portion 13 is formed has a plurality of slots 15, the first member 14 that forms the upper surface 10 </ b> A, and the groove 11. It can be manufactured by molding the second member 16 and joining them together. As shown in FIG. 4, the first member 17 in which the side wall 18 and the wall portion 13 of each groove are formed and the plate-like or sheet-like second member 19 that forms the bottom surface of each groove are integrated. You may join. Thus, there is no restriction | limiting in particular in the formation method of 10 of a base material.

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 12G are arranged in the seven grooves 11 including the grooves 11A as an example. However, 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. 5 is a cross-sectional view showing the sealing material 20 that seals the groove 11. The sealing material 20 includes a film material 21 and a heat-fusible layer 22 formed on the lower surface (one surface) of the film material 21.

  As the film material 21, 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.

The material of the heat-fusible layer 22 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 22 loses its adhesiveness again and does not affect the pretreatment material disposed in the groove 11.
The heat-fusible layer 22 may be evenly laminated on the lower surface of the film material 21 by a technique such as a co-extrusion method, or 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. It may be provided by a technique such as printing.
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 22 may be provided by a coating method in addition to the method described above. 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.

  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 22 was brought into contact with the upper surface 10A of the base material 10. When the heat-fusible layer 22 is heated by a known thermocompression bonding means in the state, the heat-fusible layer 22 and the upper surface 10A of the base material 10 are integrally fused, and the pretreatment instrument 1 is completed.

  In the pretreatment instrument 1 of the present embodiment, the sealing material 20 heat-sealed to the base material 10 has an easy peel property that can be easily peeled by a user 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.

  In each groove 11 in which the pretreatment material is disposed, the opening is formed smaller than the groove width, so that the wall portion 13 covers the upper portion of the groove even after the sealing material 20 is removed. As a result, the pretreatment material in the groove can be efficiently pretreated by suitably suppressing such a situation that the pretreatment material 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 22. It can be used. Of these, in the case of interfacial peeling, since the heat-fusible layer 22 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 22 are mixed in the groove 11. The occurrence of such a situation can be further reduced, which is preferable.

  In the pretreatment instrument of the present invention, as shown in FIG. 7, the sealing material 20 is penetrated in the thickness direction at a distance larger than the width of the opening of the groove to be sealed. Two cut portions 28 extending intermittently in the direction are formed, sandwiching the opening of the groove sealed by the cut portion 28 in plan view of the pretreatment instrument, and extending parallel to (including substantially parallel to) the groove. Thus, the sealing material 20 may be attached to the base material 10. In this way, by peeling only the area between the cut portions 28 from the substrate 10, only the desired groove (the groove 11A in the case of FIG. 7) of the plurality of grooves 11 is opened, and the other grooves are sealed. It can be kept stationary. Three or more of such cut portions may be provided at intervals larger than the width of the opening of the groove to be sealed, and the plurality of grooves may be opened independently.

  The pretreatment instrument 1 of the present embodiment configured as described above will be described in more detail using examples. In the following examples, no pretreatment material was disposed in the groove, and water was used as an alternative substance.

Example 1
The first member 14 and the second member 16 constituting the substrate 10 were formed by injection molding using an acrylic resin. The dimensions of the second member 16 were 70 millimeters (mm) in width, 70 mm in length, 10 mm in thickness, and 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. Next, the first member 14 is formed with six openings having a width of 70 mm, a length of 70 mm, a thickness of 2 mm, and a width of 3.5 mm and a length of 38 mm at positions that fit on the grooves of the second member 16. Was made. The first member 14 and the second member 16 were integrally joined with a UV curable resin to obtain the substrate 10.

  The sealing material 20 was produced as follows. A film material 21 made of two layers of polyethylene terephthalate (PET) and polyethylene (PE), a film material (manufactured by Jay Film Co., Ltd.) formed by extrusion lamination of a polyethylene-based heat-fusible layer 22 on the PE side surface. The trade name SMXZH44) was used.

  One of the grooves 11 formed in the base material 10 is filled with water, and the heat-fusible layer 22 of the sealing material 20 is directed toward the upper surface 10A of the base material 10 at 150 ° C. for 1 second under pressure. Fusion was performed. As a result, the groove 11 was suitably sealed by the sealing material 20, and the pretreatment instrument 1 was completed.

(Example 2)
Example 2 is an example having the above-described cut portion.
The sealing material 20 prepared in Example 1 was provided with a pair of perforated cut portions 28 that penetrated the sealing material and extended intermittently in parallel with an interval of 10 mm. One of the grooves 11 formed in the base material 10 is filled with water, and the sealing material 20 is positioned on the base material 10 so that the pair of cut portions 28 sandwich the opening of the groove. And pressure fused. 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, the fused sealing material 20 has an easy peel property, and the user easily peels the region between the cut portions 28 along the cut portions 28 from the base material 10 by hand. I was able to. At the time of peeling, the water filled in the groove was not scattered.

  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.

In addition, since the opening is formed narrower than the groove width in at least one of the grooves in which the pretreatment material is disposed, the pretreatment material filled in the groove is scattered during or after opening the groove. Volatilization, leakage, etc. can be suitably suppressed.
Furthermore, by forming the cut portion in the sealing material, only the groove covered with the region between the cut portions 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 22 on one surface, and the sealing material 20 seals the groove 11 when the heat-fusible layer 22 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. Furthermore, since the sealing material 20 has an easy peel property and the adhesive substance does not remain on the upper surface of the base material when it is peeled off, a pretreatment material such as a rare reagent at the time of the pretreatment becomes the adhesive substance. The risk of adhesion can be greatly reduced.

  The embodiments of the present invention have been described above. However, the technical scope of the present invention is not limited to the above-described embodiments, and various modifications are made to each component without departing from the spirit of the present invention. It is possible to delete, delete, and combine each component.

  For example, in the above-described embodiment, an example in which the openings are narrowly formed in all the grooves has been described. However, for example, a wall is provided in the groove in which the pretreatment material that is low in the risk of scattering and volatilization is disposed because it is solid. A configuration may be adopted in which the opening is narrowed only in some of the grooves, such as not providing a portion.

  Further, in the sealing material, a peeling operation may be facilitated by providing a grip portion as a handle when the user peels at the longitudinal end of the peeled region. Hereinafter, some modified examples of the pretreatment instrument of the present invention including a sealing material provided with a gripping portion will be described.

The sealing material 20A in the modification shown in FIG. 8 is formed so that the area is larger than the upper surface of the base material 10, and the cut portion 28 extends to the outer edge. One end in the longitudinal direction of a region between the cut portions 28 that is peeled when the groove 11A or the groove 11 is opened (hereinafter sometimes referred to as a “peeling region”) is released from the other regions by the cut portion 28 and is gripped. It becomes the grip part 23 which is easy to do.
In the sealing material 20B in the modification shown in FIGS. 9A and 9B, a gripping member 24A is attached to one end in the longitudinal direction of the peeling region, and a gripping portion 24 is provided. There is no restriction | limiting in particular in the material of the holding member 24, a shape, etc., You may determine suitably.

In the sealing material 20 </ b> C in the modification shown in FIG. 10, by changing the aspect of the cut portion 28, the gripping portion 25 is formed by integrating one end in the longitudinal direction of each separation region. The user can peel all the peeling regions at once and open the groove 11A and the groove 11 by gripping the grip portion 25 and performing a peeling operation.
As described above, the long gripping member 26A is fixed so that the gripping portion capable of peeling a plurality of peeling regions at a time is connected to one end in the longitudinal direction of each peeling region, like the sealing material 20D shown in FIG. It is also possible to form the gripping portion 26.
Note that it is not always necessary to unify all the peeling regions, and the number of peeling regions that can be peeled at a time may be set as appropriate.

  Furthermore, the heat-fusible layer is formed not on the entire surface of one side of the sealing material, but only on the part to be fused in contact with the base material, and the resin of the film material is exposed for the remaining part, Further treatment may be applied to increase hydrophobicity. This is preferable because the liquid pretreatment material is less likely to adhere to the sealing material. Such a heat-fusible layer can be easily formed by printing or the like.

DESCRIPTION OF SYMBOLS 1 Pretreatment instrument 10 Base material 10A Upper surface 11 Grooves 12A, 12B, 12C, 12D, 12E, 12F, 12G Pretreatment material 20, 20A, 20B, 20C, 20D Sealing material 21 Film material 22 Heat-fusible layer 23, 24, 25, 26 Grip part 28 Cut part

Claims (4)

It is a pretreatment instrument that performs pretreatment of a sample, and is an instrument having a function of performing various pretreatment steps before and after sample introduction,
A substrate having a plurality of grooves Ru extending in the same direction and open to the upper surface is formed with a constant thickness,
A pretreatment material disposed in at least one of the grooves and used for the pretreatment;
A sealing material which is formed including a film material having a heat-fusible layer made of a heat-fusible material on one surface, and seals the groove in which the pretreatment material is disposed;
With
At least one of the grooves the pretreatment material disposed is smaller than the width direction from both sides with a wall portion projecting inward in the width direction width the groove width of the opening of the groove,
The sealing material seals the groove in which the pretreatment material is disposed by heat-sealing the heat-fusible layer to the upper surface of the base material.
The sealing material has two cut portions extending in parallel with an interval larger than the width of the opening, and the cut portion is parallel to the groove in which the pretreatment material is disposed and has an opening in a plan view. It is heat-sealed to the base material so as to sandwich it,
Only the area | region between the said cut parts among the said sealing materials can be peeled from the said base material, and the peeling aspect of the said sealing material is made into interface peeling, The pretreatment instrument characterized by the above-mentioned . 2. The pretreatment instrument according to claim 1 , wherein in the sealing material, a region between the cut portions includes a grip portion for a user to grip at an end portion in a longitudinal direction.   The pretreatment device according to claim 1, wherein the heat-fusible layer is formed only at a portion that contacts the upper surface of the base material. The pretreatment instrument according to any one of claims 1 to 3, which has a function of performing various pretreatment steps before and after the sample introduction with respect to the first chip used for two-dimensional electrophoresis .

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