JP5315278B2 - 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 includes 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, each groove is sealed with a sealing material until use. This sealing material is mainly intended to prevent contamination of the groove or the pretreatment material sealed in the groove, or prevent moisture ingress when assuming integral packaging with the second-dimensional gel.
Usually, a film or the like coated with a sticky adhesive on one side is often used as the above-mentioned sealing material, but the adhesion of the reagent to the adhesive or the exudation of the adhesive component to the reagent solution, etc. Therefore, there is a concern that the result of electrophoresis is affected.

  The present invention has been made in view of the above circumstances, and an object of the present invention is to provide a pretreatment instrument that can be easily opened and used during use by reducing the influence on the pretreatment material and sealing the inside of the groove. And

The present invention relates to 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, and one surface. A film material having a heat-fusible layer made of a heat-fusible material, a recess provided in at least one of the plurality of grooves, and a composition different from that of the pretreatment material A second pretreatment material disposed in the recess, and a second film material that is partially fixed to the film material and seals the recess in the groove. The fusible layer is thermally fused to the upper surface of the base material to seal the groove in which the pretreatment material is disposed , and the second film material includes the film material for sealing the groove. By peeling off, the second film material seals the groove. When the film material is peeled off, the sealing of the concave portion is released in the same step as the peeling of the film material, and the concave portion and the groove are mixed so that the pretreatment material and the second pretreatment material are mixed. It is characterized by making it communicate.
In addition, the second film material may have a display unit indicating at least one of temperature and humidity in the groove.

  The heat-fusible layer may be selected from polyolefin-based, polypropylene-based barrier coextrusion systems, and polyolefin-based polymer alloys modified with vinyl polymers.

  The heat-fusible layer may include a heat seal lacquer composition and may be formed on one surface of the film material by coating.

  When the film material is peeled from the base material, the pretreatment device of the present invention may be peeled off by interfacial peeling.

  According to the pretreatment instrument of the present invention, the effect on the pretreatment material can be reduced to seal the inside of the groove, and the pretreatment instrument can be easily opened and used during use.

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 film material in the pretreatment instrument. It is a figure which shows the completed said pre-processing instrument. It is the elements on larger scale which show the modification of the same pretreatment instrument. It is the elements on larger scale which show the modification of the same pretreatment instrument. It is sectional drawing which shows the groove | channel in the pretreatment instrument of 2nd Embodiment of this invention. It is sectional drawing which shows the groove | channel in the modification of the embodiment. It is sectional drawing which shows the groove | channel in the modification of the embodiment.

Hereinafter, a pretreatment instrument according to a first embodiment of the present invention will be described with reference to FIGS. 1 to 6.
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 film material for sealing the groove of the substrate.

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 film 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.

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 12I are arranged in the nine 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. 3 is a diagram showing the film material 20 that seals the groove 11. A heat-fusible layer 21 containing a heat-fusible material is formed on one surface of the film material 20.
As the film material 20, 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 21 does not exhibit adhesiveness at room temperature, 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 21 loses its adhesiveness again and does not affect the pretreatment material disposed in the groove 11.

The heat-fusible layer 21 may be evenly laminated on the film material 20 by a method such as a co-extrusion method, or only a part fused on one surface, for example, the upper surface 10A of the substrate 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 21 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.

  The film material 20 configured as described above is cut into an appropriate size according to the base material 10 on which the pretreatment material is disposed, and the heat-fusible layer 21 is in contact with the upper surface 10A of the base material 10 When the heat-fusible layer 21 is heated by a known thermocompression bonding means, the heat-fusible layer 21 and the upper surface 10A of the base material 10 are fused together to complete the pretreatment instrument 1 shown in FIG. .

  The film 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 arranged is sealed to suitably prevent the entry of foreign matters from the outside and evaporation, scattering, alteration, etc. of the pretreatment material. Is easily peeled off and can be preferably used for pretreatment of the first chip.

  The peeling mode of the film material 20 is divided into a plurality of types such as cohesive peeling, delamination, and interfacial peeling depending on the material of the heat-fusible layer 21. Is possible. Among these, in the case of interfacial peeling, since the heat-fusible layer 21 does not remain on the substrate 10 side after the film material 20 is peeled, fine fragments or the like of the heat-fusible layer 21 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 film material, and no pretreatment material was disposed in the groove.
Example 1
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. As the film material 20, a film material (J-Film Co., Ltd.) formed by extrusion lamination of a polyethylene-based heat-fusible layer 21 on the PE-side surface of a film composed of two layers of polyethylene terephthalate (PET) and polyethylene (PE). ), Trade name SMXZH44).

  The top surface 10A of the base material 10 is covered so that the groove 11 is sealed with the film material 20 with the heat-fusible layer 21 of the film material 20 facing the top surface 10A of the base material 10, and is applied under conditions of 150 ° C. for 1 second. As a result of the pressure heat fusion, the groove 11 was suitably sealed by the film material 20. The fused film material 20 had easy peel properties and could be easily peeled off by the user.

(Example 2)
The same substrate 10 as in Example 1 was used. As a film material 20, a heat seal lacquer composition (made by Toyo Ink Mfg. Co., Ltd., trade name: PP fine seal) using EVA as a base resin is coated on one side of a polypropylene (PP) film material, and then heat-melted. What formed the adhering layer 21 was used.

  As a result of pressurizing and fusing the film material 20 to the base material 10 under the same conditions as in Example 1, the grooves 11 were suitably sealed by the film material 20. The film 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 film material 20 in a state where the necessary pretreatment material is disposed in the groove 11 formed in the base material 10. doing. Accordingly, the film material 20 can be stored without any problem until the time of use, and at the time of use, the film material 20 can be easily peeled off from the base material 10 and used for pretreatment, which greatly improves the convenience for the user. it can.

  Moreover, since the film material 20 is equipped with the heat-fusible layer 21 on one surface and the film material 20 seals the groove | channel 11 when the heat-fusible layer 21 is heated, the adhesion which always exhibits adhesiveness The groove 11 can be sealed without using an agent 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 film material 20 has an easy peel property and the adhesive substance does not remain in the pasting part of the base material when it is peeled off, a pretreatment material such as a rare reagent during the pretreatment becomes the adhesive substance. The risk of adhesion can be greatly reduced.

  In the present embodiment, an example in which all the grooves are sealed using one film material 20 having a size covering the entire upper surface of the substrate 10 has been described, but instead of this, any one or more By using one or a plurality of film materials cut to a size that covers the grooves, only the grooves in which the pretreatment material is disposed may be sealed with the film material.

  Further, in the film material 20, the heat-fusible layer 21 may be formed only at a position facing the upper surface 10 </ b> A of the base material 10 as in the modification shown in FIG. 5. In this case, the contact between the pretreatment material in the groove and the material of the heat-fusible layer 21 is reduced, and the pretreatment material can be prevented from being altered more suitably. When the heat-fusible layer 21 is formed in such a shape, the above-described coating method is suitable.

  Furthermore, as in the modification shown in FIG. 6, an easily breakable portion 13 such as a perforation or a slit extending in parallel with the groove 11 may be formed in the film material 20. In this case, since only specific grooves can be opened, the film material that seals each groove can be removed at an optimal timing, and a more suitable pretreatment can be performed.

  Next, a second embodiment of the present invention will be described with reference to FIGS. The difference between the pretreatment instrument 31 of the present embodiment and the pretreatment instrument 1 of the first embodiment is that a second member is provided. In the following description, components that are the same as those already described are assigned the same reference numerals and redundant description is omitted.

  FIG. 7 is a cross-sectional view showing the groove 32 in the base material 10 of the pretreatment instrument 31. The bottom surface 32 </ b> A of the groove 32 is formed as a concave portion 33 having a part deeper. In the recess 33, for example, a reagent 41 as a pretreatment material is arranged and sealed with a second film material (second member) 35.

  Similar to the film material 10, the second film material 35 has a heat-fusible layer 36 on one surface, and the heat-fusible layer 36 is heat-sealed to the bottom surface 32 </ b> A around the recess 33 to form the concave portion 33. The reagent 41 is sealed.

  A reagent 42 which is a pretreatment material used by being mixed with the reagent 41 is arranged in a region between the second film material 35 and the film material 20 in the space in the groove 32. One end portion 35A of the second film material 35 is firmly connected to the film material 20 by adhesion, welding, or the like.

  In the pretreatment instrument 31 configured as described above, when the user peels off the film material 20 from the base material 10, the sealing of the groove 32 is first released. When the user further peels off the film material 20, the end portion 35 </ b> A of the second film material 35 is pulled away from the concave portion 33, and the second film material 35 is peeled off from the bottom surface 32 </ b> A of the groove 32 to release the sealing of the concave portion 33. The

  When the sealing of the concave portion 33 is released, the reagent 41 and the reagent 42 are mixed and ready for pretreatment. The user appropriately agitates the reagents 41 and 42 as needed, and so on, soaks the first chip in the groove 32 and performs necessary pretreatment and the like.

  In the pretreatment instrument 31 of the present embodiment, since the groove 32 has the concave portion 33 and the concave portion 33 is sealed by the second film material 35, the same groove without mixing two kinds of pretreatment materials. It can seal in the state arrange | positioned to.

  Further, since the end portion 35 </ b> A of the second film material 35 is firmly connected to the film material 20, the second film material 35 can be peeled from the substrate 10 at the same time only by the operation of peeling the film material 20. . Therefore, it is possible to easily and simultaneously perform the groove sealing release operation and the pretreatment material preparation operation.

  In the present embodiment, an example in which one concave portion 33 is formed in the groove 32 has been described. However, a plurality of concave portions are formed in the groove 32, and one groove is prepared in a state where three or more kinds of pretreatment materials can be prepared at the time of use. It may be placed and sealed inside.

Moreover, although this embodiment demonstrated the example which seals the recessed part 33 provided in the groove | channel 32 with the 2nd film material 35 as a 2nd member, the aspect of a 2nd member is not limited to this but various It can be changed.
For example, as in the modification shown in FIG. 8, the second member 36 may include a display unit 36 </ b> A that displays temperature, humidity, and the like with colors, segments, characters, and the like. As a result, the user can easily know the temperature and humidity in the groove 11, so that after the pretreatment device is taken out from the cold storage location, condensation in the groove 11 is avoided or the pretreatment material is prepared at the time of use. The film material 20 can be peeled at a more suitable timing by aiming at an optimal temperature or the like. In addition, although it is possible to provide such a display part 36A in the film material 20, the direction provided in the second film material arranged in the groove 32 more accurately reflects the temperature in the groove 11 and the like. This is preferable.

  Moreover, like the modification shown in FIG. In the case where a pretreatment material (for example, solid reagent 43) that is weak against moisture or the like is disposed in the groove 11, a desiccant 37 or the like may be disposed in the groove 11 as the second member. Also in this case, since both the second members can be removed by peeling the film material 20, the quality of the reagent 43 and the like can be suitably maintained, and the preparation work can be performed more easily.

  As mentioned above, although one embodiment of the present invention has been described, the technical scope of the present invention is not limited to the above-described embodiment, and combinations of the constituent elements of each embodiment are possible without departing from the spirit of the present invention. It is possible to change, to make various changes to each component, or to delete.

DESCRIPTION OF SYMBOLS 1, 31 Pretreatment instrument 10 Base material 11, 11A, 32 Grooves 12A, 12B, 12C, 12D, 12E, 12F, 12G, 12H, 12I Pretreatment material 20 Film material 21 Heat-fusible layer 35 Second film material ( Second member)
36 Second member 37 Desiccant (second member)
41, 42, 43 Reagent (Pretreatment material)

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 film material having a heat-fusible layer made of a heat-fusible material on one surface;
A recess provided in at least one of the plurality of grooves;
A second pretreatment material having a composition different from that of the pretreatment material and disposed in the recess;
A second film material that is partly fixed to the film material and seals the recess in the groove;
With
The film material seals the groove in which the pretreatment material is disposed by thermally fusing the heat-fusible layer to the upper surface of the base material .
Said 2nd film material cancels | releases sealing of the said recessed part in the same process as peeling of the said film material, when the said film material which seals the said groove | channel is peeled, The said pre-processing material and said 2nd pre-processing The recess and the groove communicate with each other so that the material is mixed.
A pretreatment instrument characterized by that.   The pretreatment instrument according to claim 1, wherein the second film material has a display unit indicating at least one of temperature and humidity in the groove. The pretreatment device according to claim 1 or 2 , wherein the heat-fusible layer is selected from a polyolefin-based, a polypropylene-based barrier coextrusion system, and a polyolefin-based polymer alloy modified with a vinyl polymer. The pretreatment instrument according to claim 1 or 2 , wherein the heat-fusible layer contains a heat seal lacquer composition and is formed on one surface of the film material by coating. The pretreatment instrument according to any one of claims 1 to 4 , wherein when the film material is peeled from the base material, the film material is peeled off by interfacial peeling.

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