JP2012157267A - Plate member having fine pattern - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a fine structure favorably dealing with a well passing phenomenon of beads while considering "rigidity" and "beads housing properties" or the like.SOLUTION: A plate member having a fine pattern constitute of a plurality of same shaped recessed parts includes at least a first side wall part wherein a side wall surface forming the recessed parts are located on opening surface sides of the recessed parts and a second side wall part located on a bottom surface side of the first side wall part. An inclination angle α1 (the inclination angle of solid part side of the plate member) of the first side wall part to the horizontal surface is smaller than an inclination angle α2 (the inclination angle of the solid part side of the plate member).

Description

  The present invention relates to a plate member having a fine pattern. More specifically, the present invention relates to a plate member having a fine pattern composed of a plurality of concave portions on the order of microns.

  Microstructures below the micron order are generally used in a wide range of fields such as MEMS (Micro Electro Mechanical System) and μ-TAS (Micro Total Analysis System), and technologies such as machining and photolithography are applied. ing.

  In recent years, such fine structure fabrication technology has also been applied in the fields of chemistry, medical science, and bioscience, and the fabricated microstructure is separated, immobilized, analyzed, extracted, and purified of substances or cells. Or it is used suitably for various processes, such as reaction. Such a fine structure generally has a flat plate (or chip) plate form or a laminated form formed by laminating these, and includes, for example, a well plate, a microtiter plate, and the like. They are called chemical chips, biochips, DNA (deoxyribonucleic acid) chips, microarray chips, and the like.

  For example, taking a microarray chip that can be used for DNA analysis as an example, in order to process and analyze a large amount of information at a time, many thousands of hundreds of thousands of “concave portions” or It is necessary to create a “dent”. Such “recesses” or “recesses” are generally called wells, and can contain microbeads, liquids (reaction liquids), etc. (for example, if the volume of each well is constant, a prescribed volume of reaction liquid is Can be stored in wells).

  Since it is necessary to provide a large number of wells in this way, it is more suitable to fabricate by batch exposure using photolithography rather than machining each fine structure one by one. In other words, a resist master is manufactured using photolithography, a stamper is manufactured from the resist master, and then a transfer structure such as injection molding or hot embossing using the stamper as a mold is applied to collect the microstructures at once. And have gained.

Product information of Sumitomo Bakelite Co., Ltd. (Product name: Multiplate for culture) [online], [Searched on December 17, 2010], Internet <URL: http://www.sumibe.co.jp/sumilon/plate .html>

  As a result of intensive studies on the structure such as the well plate described above, the inventors of the present application have found that there is a problem peculiar to the “fine structure” at the time of use. Specifically, when a mixture of microbeads and a liquid (reaction solution) was applied to a fine structure, a phenomenon was found in which beads that should have settled into the well pass by without being collected in the well. (Hereinafter also referred to as “well passage phenomenon”).

  The “well passage phenomenon” occurs when microbeads pass over the flat structure between the wells. The individual microbeads are suspended and dispersed in the liquid, and the movement of the beads becomes very complicated depending on the flow of the liquid. Therefore, it is difficult to control the path of the beads passing through the flat portion between the wells.

  A possible solution is to narrow the pitch of the well array and reduce the flat part, but the side wall part that forms each well becomes thin, so the rigidity of the microstructure is reduced, resulting in a structurally weak plate. End up. Since the well size is on the order of microns, the structure itself is not rigid in the first place, and therefore it is difficult to reduce the distance between the wells. That is, the dimension of the flat portion cannot be simply reduced.

  Further, in order to increase the bead accommodation rate in the well, a method of simply increasing the well diameter can be considered. However, increasing the well diameter also leads to thinning of the side wall portions forming each well, so that the plate structure becomes weak and the rigidity decreases. If the well diameter is increased without changing the side wall width, it is necessary to change the well pitch itself, and the design of the entire manufacturing apparatus needs to be changed.

  Furthermore, when the well diameter is increased, the volume of the well itself may increase more than necessary, and the number of microbeads accommodated in the well may exceed the expected number. As a result, it is necessary to change various specifications such as changing the relationship between the reaction solution and the bead surface area from the initial plan / plan. In particular, since the number of microbeads accommodated in the well is determined in advance and the volume is determined accordingly, in this case, the well diameter cannot be easily increased.

  The present invention has been made in view of the above circumstances. That is, an object of the present invention is to provide a microstructure that can suitably cope with the phenomenon of beads passing through wells while taking into account “rigidity” and “bead accommodation characteristics”.

In order to solve the above problem, the present invention is a plate member having a fine pattern composed of a plurality of concave portions having substantially the same shape,
The side wall surface forming the concave portion is composed of at least a “first side wall portion located on the opening surface side of the concave portion” and a “second side wall portion located on the bottom surface side of the concave portion”,
The inclination angle α1 of the first side wall with respect to the horizontal plane (inclination angle on the solid part side of the plate member) is smaller than the inclination angle α2 of the second side wall part (inclination angle on the solid part side of the plate member). A plate member is provided.

  One of the features of the present invention is that the side wall surface of the recess is composed of at least two portions having different inclinations, and the first side wall portion located on the upper side (opening surface side) is further on the lower side (bottom surface side). It is that the inclination is gentler than that of the second side wall located at (see FIG. 1). As a result, a microstructure capable of efficiently guiding the microbeads into the recesses (wells) while suppressing an increase in the recess volume / well volume and a decrease in the rigidity of the structure is realized.

  In this specification, the “fine pattern” is a pattern constituted by a plurality of continuous fine recesses. Since a plurality of continuous fine recesses result in a fine uneven shape, it can be said that the “fine pattern” is a pattern composed of fine uneven shapes. In the present invention, such a “fine pattern” is a dimension (width / height / width) of a “fine concave portion (or fine dent)” or “fine convex portion (or fine bulge)” constituting the fine concavo-convex shape. It means that the dimension (depth, etc.) is in the micron order (in the range of 0.1 μm to 1000 μm, for example, 1 μm to 600 μm). For example, when the plate member corresponds to a well plate or the like, the “fine recess” or “fine recess” often forms a well shape, whereas when the plate member corresponds to a microchemical chip or the like, the “fine recess” or “Fine depressions” often form channel shapes (groove shapes) or channel shapes.

  Further, the “plate member” in the present specification refers to a “plate-like” or “chip-like” member used in the fields of chemistry, medical science, bioscience, and the like. Such a plate member may generally have a thickness of, for example, about 100 μm to 50 mm.

  In a preferable aspect, the inclination angle α1 of the first side wall portion is smaller by 5 ° to 60 ° than the inclination angle α2 of the second side wall portion. The inclination angle itself (α1) of the first side wall portion is preferably in the range of 30 ° to 70 °, for example. On the other hand, the inclination angle itself (α2) of the second side wall portion is preferably in the range of 75 ° to 90 °, for example.

In another preferable aspect, among the width dimensions of the respective recesses, the width dimension L _{opening portion} located closest to the opening surface when viewed along the vertical direction (that is, the recess depth direction) is the largest. In particular, it is preferable that the “width dimension L _opening located closest to the opening surface” has a length equal to or shorter than the pitch of the plurality of recess arrays.

  As a form of the first side wall part and the second side wall part, for example, at least one of the first side wall part and the second side wall part may have a curved surface shape. Alternatively, at least one of the first side wall part and the second side wall part may have a planar shape. In other words, both the first sidewall portion and the second sidewall portion may have a curved surface shape or a planar shape, or the first sidewall portion has a curved surface shape and the second sidewall portion has a planar shape. The first side wall portion may have a planar shape and the second side wall portion may have a curved surface shape.

  In a preferred aspect, the boundary portion between the first side wall portion and the second side wall portion is angular. That is, a corner is formed at the boundary between the first side wall and the second side wall. In this case, for example, the first side wall portion may have a curved shape, while the second side wall portion may have a planar shape.

  In another preferable aspect, a boundary portion between the first side wall portion and the second side wall portion is smooth. That is, the boundary portion between the first side wall portion and the second side wall portion is continuously and smoothly formed without being angular.

  In still another preferred aspect, each recess has a third side wall portion located further on the bottom surface side than the second side wall portion. In this case, the inclination angle α3 of the third side wall portion with respect to the horizontal plane (inclination angle on the solid portion side of the plate member) is larger than the inclination angle α1 of the first side wall portion and the inclination angle α2 of the second side wall portion. It is preferable.

  The bottom surface of each recess may have any form. For example, the bottom surface of the concave portion may be not only a flat surface but also a conical surface or a hemispherical surface (or a bottom surface having an arc shape when viewed in a cross section of the concave portion). Moreover, in each recessed part, the side wall part different from said 1st-3rd side wall part etc. may be provided additionally. For example, the concave portion may further include a “vertical side wall portion positioned further on the opening surface side than the first side wall portion (the inclination angle of the vertical side wall portion with respect to the horizontal plane is 90 °)”.

  In this invention, the metal metal mold | die used in order to shape | mold the above-mentioned plate member is also provided. Such metal molds preferably have the form of rolls or flat plates. Such metal molds are Ni (nickel), Ag (silver), Au (gold), Bi (bismuth), Cd (cadmium), Co (cobalt), Cr (chromium), Cu (copper), Fe (Iron), In (indium), Pd (palladium), Pt (platinum), Ru (ruthenium), Sn (tin) and at least one metal selected from the group consisting of Zn (zinc) .

  In the plate member of the present invention, the microbead can be guided to the center of the well while maintaining the rigidity of the fine pattern, and the bead can be efficiently accommodated in the well. In other words, the plate member of the present invention suitably copes with the “well passage phenomenon” without substantially impairing its rigidity.

In particular, even if the fine concave portions are arranged at high density in the plate member (for example, even if the width dimension L _{opening portion} located closest to the opening surface in each concave _portion has a length equal to or less than the pitch of the concave portion arrangement) ) Since the solid portion of the plate between the adjacent concave portions is thick, the rigidity of the plate member is not substantially impaired.

  In this way, the plate member of the present invention has a high density of fine concave portions and a wide opening portion of the concave portion (well) due to the inclination angle of the first side wall portion while maintaining rigidity. Microbeads can be effectively guided into the recesses (see FIG. 2). In addition, even though the opening of the recess (well) is so wide, the recess width dimension becomes smaller toward the bottom of the recess (well), so that the beads once accommodated in the recess are pulled out. It is difficult.

  More specifically, it is also possible to design to accommodate only a predetermined number (for example, one) of beads in each recess by appropriately adjusting the width dimension of the bottom side portion of the recess and the inclination angle of the second side wall. . In consideration of this point and the inclination angle of the first side wall, a plate member capable of holding an appropriate amount of liquid in the recess while controlling a predetermined number of beads in the recess is realized. it can.

FIG. 1 is a perspective view and a cross-sectional view schematically showing a concave shape (well shape) according to the present invention. FIG. 2 is a view schematically showing an aspect in which microbeads are captured in the recesses (wells) according to the present invention. FIG. 3 is a perspective view schematically showing a plate member and a recess (well) according to the present invention. FIG. 4 is a perspective view, a top view, and a supplementary view schematically showing an aspect of a recess (well) in FIG. 3. FIG. 5 is a diagram for explaining the “tilt angle”. FIG. 6 is a diagram for explaining the “inclination angle” (particularly the inclination angle of the side wall portion forming the curved surface). FIG. 7 is a perspective view schematically showing an aspect of a plate member and a recess (well) according to the present invention. FIG. 8 is a perspective view, a top view, and a supplementary view schematically showing the aspect of the recess (well) in FIG. 7. FIG. 9 is a perspective view schematically showing a plate member and a recess (well) according to the present invention. FIG. 10 is a perspective view, a top view, and a supplementary view schematically showing the aspect of the recess (well) in FIG. 9. FIG. 11 is a perspective view schematically showing an aspect of a plate member and a recess (well) according to the present invention. FIG. 12 is a perspective view, a top view, and a supplementary view schematically showing the aspect of the recess (well) in FIG. 11. FIG. 13 is a perspective view schematically showing a plate member and a recess (well) according to the present invention. FIG. 14 is a perspective view, a top view, and a supplementary view schematically showing the aspect of the recess (well) in FIG. 13. FIG. 15 is a perspective view schematically showing a plate member and a recess (well) according to the present invention. FIG. 16 is a perspective view, a top view, and a supplementary view schematically showing an aspect of the recess (well) in FIG. 15. FIG. 17 is a perspective view, a top view, and a supplementary view schematically showing an embodiment of a recess (well) according to the present invention. FIG. 18 is a perspective view, a top view, and a supplementary view schematically showing an aspect of a recess (well) according to the present invention. FIG. 19 is a diagram schematically showing a recess (well) of Comparative Example 1 as a conventional technique. 20 is a plan view schematically showing the chromium mask used in Comparative Example 1. FIG. FIG. 21 is a plan view schematically showing the multi-tone mask used in the first embodiment. FIG. 22 is an electron micrograph of a recess (well) formed in Example 1. FIG. 23 is an electron micrograph of a recess (well) formed in Example 2. FIG. 24 is an electron micrograph of the recesses (wells) formed in Example 4. FIG. 25 is an electron micrograph of a recess (well) formed in Example 4. FIG. 26 is an electron micrograph of a recess (well) formed in Example 4. FIG. 27 is an electron micrograph of a recess (well) formed in Example 4. FIG. 28 is a diagram showing a well shape that is a target of the structural simulation in the “confirmation test of the rigidity characteristics of the plate member”. FIG. 29 is a graph showing a simulation result of the displacement amount of the well side wall when a load is applied.

  Hereinafter, the present invention will be described in more detail with reference to the drawings.

<General characteristics of plate members>
As shown in FIG. 3, the plate member 100 of the present invention has a plurality of recesses 50 formed in its body portion 60. In particular, in the present invention, the side wall surface of each recess is at least composed of “a first side wall portion 10 positioned on the opening surface side of the recess” and “a second side wall portion 20 positioned on the bottom surface side of the recess”. As illustrated, the first side wall 10 and the second side wall 20 are continuously provided along the depth direction of the recess. The inclination angle α1 of the first side wall with respect to the horizontal plane (inclination angle located on the solid part side of the plate member) is the inclination angle α2 of the second side wall with respect to the horizontal plane (inclination located on the solid part side of the plate member). Smaller than the angle). That is, the inclination of the first side wall portion positioned on the relatively upper side (opening surface side) is gentler than that of the second side wall portion positioned on the lower side (bottom surface side).

  The material of the plate member 100 of the present invention (that is, the material of the body portion 60) may be resin, glass, silicon, metal, or the like. For example, for resin, there is no particular limitation, but cycloolefin resin (norbornene resin), cycloolefin copolymer resin, polycarbonate (PC), amorphous polyolefin, polydimethylsiloxane, phenol resin, epoxy resin, melamine resin , Urea resin, unsaturated polyester resin, alkyd resin, polyurethane, polyimide, polyethylene, polypropylene (PP), polyvinyl chloride, polystyrene, polyvinyl acetate, polyvinyl ether, polytetrafluoroethylene, acrylonitrile resin, butadiene resin, styrene resin, Acrylic resin, poly (meth) acrylic acid, poly (meth) acrylic acid ester, polyamide, polyacetal, modified polyphenylene ether, polybutylene terephthalate, At least one polymer selected from the group consisting of reethylene terephthalate, polyphenylene sulfide, polysulfone, polyether sulfone, polyvinyl alcohol, amorphous polyarylate, liquid crystal polymer, polyetheretherketone, poamideimide, polyallylamine, and polyethyleneimine. It may be a resin.

  The dimensions of the plate member 100 of the present invention can vary depending on the application. For example, taking a well plate-shaped plate member 100 as shown in FIG. 3 as an example, the device dimensions (L, H, W) are L = 5-300 mm, H = 0.2-50 mm, W = 10-300 mm. Can be a degree.

In general, the width dimension and the depth dimension of the recess 50 are preferably larger than the diameter dimension D of the microbeads, so that an appropriate amount of liquid such as a reaction solution can be accommodated in each recess. Can do. Depending on the inclination angle of the first side wall and the second side wall, which will be described later, taking the well plate as shown in FIG. 3 as an example, for example, each recess has a range of the following width and height dimensions. have:

Width / W: 6 μm to 600 μm
· W1: 2 μm to 200 μm
・ W2: 3 μm to 300 μm

Height dimension : H: 5 μm to 400 μm
・ H1: 2 μm to 160 μm
・ H2: 3 μm to 240 μm

Although it can be said that the larger the width dimension W and the depth dimension H of the recess 50, the larger the amount of liquid necessary for the reaction and the like, which is preferable in many cases, the amount of the necessary component exceeds the amount necessary and sufficient for the reaction. If this happens, unnecessary liquid costs will be incurred. For this reason, although it depends on the form of the recess 50 (for example, the form of its side wall), the upper limit of the width dimension W and the depth dimension H of the recess 50 is preferably about 10D or less, and more preferably about 5D or less. (D: average size of microbeads). Incidentally, in the case where efficient stirring is possible by subjecting to vibration or the like, since necessary components can be moved in a wide range, larger width and depth dimensions may be used.

  The number of the recesses 50 provided in the plate member 100 of the present invention can be variously changed depending on the application. For example, in the case where many of the plurality of recesses 50 correspond to “wells”, such as well plates, microtiter plates, etc., the number of recesses (ie “wells”) is preferably 2500 to 100 million, more preferably It may be 10,000 to 50 million, more preferably 50,000 to 10 million.

  Preferably, the plate member 100 of the present invention is used with a dispersion containing “a plurality of microbeads” and “liquid” in use. Specifically, a microbead is provided in each of the recesses by providing a dispersion liquid composed of microbeads and liquid to the “recess mounting surface” of the plate member (that is, the plate surface on which the fine pattern is provided). And the liquid are accommodated together, or the dispersion is allowed to flow so that the microbeads and the liquid pass through the recess. For example, in such “accommodation”, although the sedimentation action resulting from the dead weight of the microbeads and the liquid may be used, the device may be shaken or subjected to a centrifugal treatment as necessary. . Furthermore, a liquid or gas flow may be provided, a magnetic field may be applied in the case of magnetic microbeads, or an electric field may be provided in the case of chargeable microbeads.

  The “microbead” and “liquid” will be described in detail. “Microbeads” substantially mean microparticles conventionally used in the fields of medical science and bioscience. The microbeads may be not only fine particles such as metal fine particles, polymer fine particles or glass fine particles, but also particulate matter such as cells, microorganisms, bacteria, pollen, spores and other biologically relevant particles. Such microbeads may have various shapes such as a spherical shape, an elliptical shape, a rice grain shape, a needle shape, or a plate shape. The term “spherical” as used herein refers to a shape having an aspect ratio (ratio between the maximum length and the minimum length when measured in various directions) in the range of 1.0 to 1.2. Indicates a shape having an aspect ratio in the range of 1.2 to 1.5 (excluding 1.2). In addition, “rice grain” means a shape like “rice grain” as the name suggests. From the viewpoint of promoting “accommodation in the recess”, the microbeads preferably have a spherical shape. The average size D of the microbeads is, for example, preferably about 0.01 μm to 10 mm, more preferably about 0.1 μm to 5 mm. Here, the “size” substantially means the maximum length among the lengths in all directions of the beads. The “average size” substantially means a value obtained by measuring the size of, for example, 300 beads based on a transmission electron micrograph or optical micrograph of beads and calculating the number average. On the other hand, a “liquid” is a liquid that is conventionally used with microbeads in the fields of medical science and bioscience, and is, for example, water, a solvent, a buffer, a reaction solution, or a sample solution. There is no restriction | limiting in particular in the viscosity of a liquid, The liquid should just have fluidity on the conditions when utilizing the component in a liquid. The microbead content of the dispersion liquid (that is, the dispersion liquid composed of microbeads and liquid) used for the microplate device is not particularly limited, but is about 0.01 to 50% by volume. The absolute amount or number of microbeads can vary depending on the application. For example, if the “concave portion” is 10,000 wells, the number of microbeads is preferably at least 10,000, and microbeads having an absolute amount corresponding to the number of microbeads may be included in the dispersion.

  This invention is made paying attention to the form of the recessed part (dent) in a plate member, especially the form of a recessed part side wall surface. In the present invention, as shown in FIG. 1, in each of the recesses, the side wall surface is at least composed of a first side wall portion 10 positioned on the opening surface side of the recess and a second side wall portion 20 positioned on the bottom surface side of the recess. The inclination angle α1 of the first side wall portion 10 is smaller than the inclination angle α2 of the second side wall portion 20 (in the case of looking along the circumferential direction of the concave portion, the first side wall portion or the second side wall portion). It is preferable that each inclination angle of the part is the same).

  Due to the relative inclination angle between the first side wall and the second side wall, the typical shape of the concave portion of the plate member of the present invention is more upward (as shown in FIGS. 3 and 4). The first side wall portion 10 positioned on the opening surface side is inclined more gently than the second side wall portion 20 positioned on the lower side (bottom surface side). In other words, the concave portion 50 is formed with a structure that gradually changes from the maximum length portion Lmax to the shortest length portion Lmin, whereby the opening of the maximum length portion Lmax has the widest shape (see FIG. 4). As a result, the collection rate of beads can be increased without changing the well pitch and without reducing the rigidity of the chip. In other words, the beads can be effectively guided to the center of the well when the plate member is used while maintaining the rigidity of the fine pattern of the plate member.

  Here, the “inclination angle” used in the present specification is an angle with respect to the horizontal plane Z (which may correspond to the opening surface A) as shown in FIG. It means the angle α (α1, α2,...) Located on the part side / outside (see FIG. 5A). In other words, when a plate member (particularly a device having a flat bottom surface) is placed on a horizontal surface, the outer angle formed by the “side wall surface” and the “horizontal plane Z” corresponds to the “inclination angle”. . Here, as shown in FIG. 5B, when the side wall surface is a curved surface, the “inclination angle” corresponds to the “average inclination angle”. For simplicity, the “average inclination angle” means that a “straight line connecting the upper edge and the lower edge of the side wall portion” forms the concave opening surface or the horizontal plane Z as shown in FIG. It means the angle substantially. Further, the “average inclination angle” is regarded as “an angle formed by the“ tangent line having the intermediate point S of the curved surface as a contact point ”with the opening surface of the depression or the horizontal plane Z, as shown in FIGS. 6 (a) and 6 (b). (Note that the “curved surface intermediate point S” is a curved surface point located at half the depth of the target side wall).

  The inclination angle α1 of the first side wall portion is not particularly limited, but is preferably in the range of 30 ° to 70 °, for example. In general, the smaller the inclination angle α1, the more efficiently microbeads can be guided into the recesses (wells), and it becomes easier to cope with the phenomenon of beads passing through the well, while the inclination angle α1 is more than necessary. When it becomes smaller, the number of recesses (recess density) per unit plate surface is reduced. In some cases, the inclination angle α1 of the first side wall portion may be in the range of 35 ° to 50 °.

  The inclination angle α2 of the second side wall portion is not particularly limited, but is preferably in the range of 75 ° to 90 °, for example. In general, when the inclination angle α2 is large, the bead stagnates at the side wall portion or rides on the side wall portion is reduced. Further, the larger the inclination angle α2, the more securely the beads once captured in the recess (well) can be held in the recess (well). However, when the inclination angle α2 becomes larger than necessary, the recess ( This can be difficult if the microbeads captured in the (well) are to be removed from within the recess (well). In some cases, the inclination angle α2 of the second side wall portion may be in the range of 85 ° to 90 ° (for example, about 90 °).

  In the plate member of the present invention, it is preferable that the inclination angle α1 of the first side wall portion is smaller by 5 ° to 60 ° than the inclination angle α2 of the second side wall portion. Such a difference in inclination angle may be selected as necessary. In general, the greater the difference in the angle of inclination, the greater the average thickness (solid part thickness) between adjacent recesses, which can contribute to rigidity retention, but more than necessary. If it is large, the number of recesses (concave density) provided per unit plate surface will be reduced. Therefore, the difference (α1 <α2) between “the inclination angle α1 of the first side wall portion” and “the inclination angle α2 of the second side wall portion” is more preferably about 10 ° to 55 °, and further preferably 25 ° to 50 °. It is about °.

As can be seen from the embodiment shown in FIG. 2, the “width dimension L _opening located closest to the opening surface when viewed along the vertical direction / depth direction” is the largest dimension among the width dimensions of each recess. Yes. It can be said that because of such a large size, the microbeads can be efficiently guided into the recesses (wells), and the phenomenon of beads passing through the wells can be suitably dealt with. In particular, it is preferable that the width dimension L _opening has a length equal to or less than the pitch P of the plurality of recesses. As a result, there is no overlap between the patterns, and even when they are closest, the outer edges are in contact with each other. Therefore, the collection rate can be increased efficiently without substantially changing the arrangement of the recesses (wells). Can do. In other words, the number of recesses (recess density) per plate unit area can be increased while maintaining the rigidity of the fine pattern. Further, as shown in FIG. 3 and FIG. 4 and the like, the well has a wide opening and is gradually narrowed toward the bottom, so that the beads are easily collected in the well and the beads once collected in the well Unnecessary escape by liquid flow is reduced.

<< Various aspects of recesses >>
Various other modes are conceivable as the “concave portion”. This will be described below.

(Mode in which the second side wall is a vertical surface)
The aspect in which the second side wall portion 20 is a vertical surface is shown in FIGS. Even in this aspect, the inclination angle α1 of the first side wall portion 10 is smaller than the inclination angle α2 of the second side wall portion 20. In the concave portion as shown in the drawing, the inclination angle α2 of the second side wall portion is about 90 °, whereas the inclination angle α1 of the first side wall portion is about 45 °.

  As shown in FIG. 8, in each recess 50, the first side wall 10 and the second side wall 20 both have a planar shape, and the boundary portion between the first side wall 10 and the second side wall 20. Is square, that is, the corner is out. When the boundary portion is square as described above, it is possible to further effectively reduce the phenomenon that “beads once collected in the well are unnecessarily escaped by the liquid flow”.

  Even in the plate member 100 having the concave portion shown in FIGS. 7 and 8, the microbead can be guided to the center of the well while maintaining the rigidity of the fine pattern, and the bead is efficiently accommodated in the well. Can do.

(Aspect in which first side wall portion is curved-case 1)
A mode (case 1) in which the first side wall portion 10 is curved is shown in FIGS. Also in this aspect, the inclination angle α1 of the first side wall portion 10 is smaller than the inclination angle α2 of the second side wall portion 20. In the concave portion as shown in the drawing, the inclination angle α2 of the second side wall portion is about 70 ° to about 85 °, whereas the inclination angle α1 of the first side wall portion is about 30 ° to about 50 °. It has become. In particular, the curved surface of the first side wall portion 10 is formed so as to draw an arc toward the bottom side.

  As shown in FIG. 10, in each recess 50, the first side wall portion 10 has a curved surface shape, while the second side wall portion 20 has a planar shape. Moreover, the boundary part of the 1st side wall part 10 and the 2nd side wall part 20 is angular, ie, the corner has come out. When the boundary portion is square as described above, it is possible to further effectively reduce the phenomenon that “beads once collected in the well are unnecessarily escaped by the liquid flow”.

  9 and FIG. 10, even if the plate member 100 has a concave shape, the microbead can be guided to the center of the well while maintaining the rigidity of the fine pattern, and the beads can be efficiently accommodated in the well. Can do.

(Aspect in which the first side wall is curved-case 2)
An aspect (case 2) in which the first side wall 10 is curved is shown in FIGS. Also in this aspect, the inclination angle α1 of the first side wall portion 10 is smaller than the inclination angle α2 of the second side wall portion 20. In the embodiment as illustrated, the inclination angle α2 of the second side wall portion is about 90 °, whereas the inclination angle α1 of the first side wall portion is about 30 ° to about 50 °. In particular, the curved surface of the first side wall 10 is formed to draw an arc toward the opening side.

  As shown in FIG. 12, in each recess 50, the first side wall portion 10 has a curved shape, while the second side wall portion 20 has a planar shape. Further, the boundary portion between the first sidewall portion 10 and the second sidewall portion 20 is smooth. That is, the boundary portion between the first side wall portion and the second side wall portion is continuously and smoothly formed without being angular. When the boundary portion is formed smoothly in this way, the beads can be more effectively guided to the well center.

  Even in the plate member 100 having the concave portion shown in FIGS. 11 and 12, the microbead can be guided to the center of the well while maintaining the rigidity of the fine pattern, and the bead is efficiently accommodated in the well. Can do.

(Aspect with third side wall-case 1)
The aspect (case 1) which has the 3rd side wall part 30 is shown in FIG. 13 and FIG. As can be seen from the illustrated embodiment, in this embodiment, the third sidewall 30 is provided further on the bottom side than the second sidewall 20. Even in this aspect, the inclination angle α1 of the first side wall portion 10 is smaller than the inclination angle α2 of the second side wall portion 20. The inclination angle α3 of the third side wall portion 30 is larger than the inclination angle α1 of the first side wall portion 10 and the inclination angle α2 of the second side wall portion 20 as shown in FIG. The part has a "multi-stage taper structure"). For example, a specific inclination angle of the first to third side walls is about 30 ° to about 50 °, and an inclination angle α2 of the second side wall is about 60. The inclination angle α3 of the third side wall portion is about 90 °.

  As shown in FIG. 14, in each recess 50, the first side wall part 10, the second side wall part 20, and the third side wall part 30 all have a planar shape. Further, the boundary portion between the first sidewall portion 10 and the second sidewall portion 20 is angular, and the boundary portion between the second sidewall portion 20 and the third sidewall portion 30 is also angular.

  As described above, even in the plate member 100 having the concave shape shown in FIGS. 13 and 14, the microbead can be guided to the center of the well while maintaining the rigidity of the fine pattern, and the bead is efficiently put into the well. Can be accommodated.

(Aspect with third side wall-case 2)
The aspect (case 2) which has the 3rd side wall part 30 is shown in FIG. 15 and FIG. As can be seen from the illustrated embodiment, also in this embodiment, the third sidewall 30 is provided further on the bottom surface side than the second sidewall 20. Also in this aspect, the inclination angle α1 of the first side wall portion 10 is smaller than the inclination angle α2 of the second side wall portion 20. The inclination angle α3 of the third side wall portion 30 is larger than the inclination angle α1 of the first side wall portion 10 and the inclination angle α2 of the second side wall portion 20 as shown in the drawing (that is, the side wall portion of the recess). Is a “multi-step taper structure”). If the specific inclination angle of the first to third side wall parts is exemplified, the inclination angle α1 of the first side wall part is about 50 ° to about 60 °, and the inclination angle α2 of the second side wall part is about 60. The inclination angle α3 of the third side wall portion is about 90 °.

  As shown in FIG. 16, in each recessed part 50, while the 2nd side wall part 20 and the 3rd side wall part 30 have comprised planar shape, the 1st side wall part 10 has comprised curved surface shape. Further, the boundary portion between the first sidewall portion 10 and the second sidewall portion 20 is angular, and the boundary portion between the second sidewall portion 20 and the third sidewall portion 30 is also angular.

  As described above, even in the plate member 100 having the concave portion shown in FIGS. 15 and 16, the microbead can be guided to the center of the well while maintaining the rigidity of the fine pattern, and the bead is efficiently put into the well. Can be accommodated.

(Aspect with a vertical side wall located on the opening surface)
A mode provided with the vertical side wall located on the opening surface is shown in FIG. As can be seen from the illustrated embodiment, in this embodiment, the vertical side wall portion 40 is provided on the opening surface side further than the first side wall portion 10. Even in this aspect, the inclination angle α1 of the first side wall portion 10 is smaller than the inclination angle α2 of the second side wall portion 20. In the illustrated recess, the inclination angle α2 of the second side wall portion 20 is about 90 °, whereas the inclination angle α1 of the first side wall portion 10 is about 30 ° to about 50 °. . And the inclination | tilt angle of the vertical side wall part 40 is about 90 degrees as the name suggests. Even in the embodiment shown in FIG. 17, the microbead can be guided to the center of the well while maintaining the rigidity of the fine pattern, and the bead can be efficiently accommodated in the well.

(Aspect in which the bottom surface of the concave portion forms a conical surface or a hemispherical surface)
FIG. 18 shows an embodiment in which the bottom surface of the concave portion forms a conical surface or a hemispherical surface. As can be seen from the illustrated embodiment, in this embodiment, the bottom surface of the concave portion forms a substantially conical surface (FIG. 18A) or a substantially hemispherical surface (FIG. 18B). Even in this aspect, the inclination angle α1 of the first side wall portion 10 is smaller than the inclination angle α2 of the second side wall portion 20. In the illustrated recess, the inclination angle α2 of the second side wall portion 20 is about 90 °, while the inclination angle α1 of the first side wall portion 10 is about 20 ° to about 50. Even in the embodiment as shown in FIG. 18, the microbead can be guided to the center of the well while maintaining the rigidity of the fine pattern, and the bead can be efficiently accommodated in the well.

<< Utilization Method of the Present Invention >>
Next, how to use the plate member of the present invention will be described. As a typical usage method of the plate member of the present invention, the “mixture of the microbead 70 and the liquid 80” is allowed to flow into the concave portion of the plate member, and the microbead and the liquid are accommodated in each concave portion 50. Thereby, desired processing such as separation, immobilization, analysis, extraction, reaction or mixing of the target substance is performed (see FIG. 2).

Since the specific gravity of the liquid is generally around 1, beads having a specific gravity greater than 1.02 are preferably used as the sedimenting beads (here, the “specific gravity” is 4 ° C. water [about 1.0 g / cm ³ ] based on this). In addition, as the beads that can be easily moved according to the inclination, beads having a specific gravity of 2 are preferably used. Most preferred are beads with a specific gravity greater than 3.5. As the specific gravity increases, the inertia increases and the overall weight of the device including the beads also increases. Therefore, the preferable upper limit of the specific gravity of the beads is 10 or less.

  If a substance to be examined, a substance related to a reaction, or the like is fixed to the microbeads as necessary, the movement of the substances in the plate member becomes possible as the beads move. In such an embodiment, it is preferable to fix a substance, a functional group, or the like capable of fixing or adsorbing a substance to be examined or a substance related to a reaction to the bead surface. Substances and functional groups to be immobilized include inorganic substances such as silica, gold and hydroxyapatite capable of binding nucleic acids and proteins, functional groups capable of chemically binding these, avidin or biotin capable of binding to each other, specifically Examples thereof include an antibody capable of binding an antigen, an antigen capable of specifically binding an antibody, and the like.

In the following, we will touch on various treatment methods with an eye on actual applications. When the plate member of the present invention is used, separation, immobilization, analysis, extraction or reaction of the target substance can be suitably performed, but the well plate was used for the purpose of exemplifying specific embodiments relating to these processes. The analysis / extraction and purification of the target substance and the separation / detection / screening will be explained.

Analysis / extraction / purification of target substance: In “analysis of target substance”, for example, a sample liquid containing or possibly containing the target substance or a mixture of the sample liquid and other liquids (hereinafter referred to as a sample-containing liquid), Contact a bead on which antigen, antibody, nucleic acid, etc. that specifically binds to the target substance are immobilized on the surface in contact with the well, and after giving conditions for sufficient binding of the target substance to the bead surface, remove or wash away the sample liquid. Repeat the washing as necessary. Beads filled with a solution containing "labeling substance" that binds another antigen, antibody, nucleic acid, etc. that specifically binds to the target substance, and a fluorescent substance, chromogenic substrate, luminescent substrate, magnetic substance, etc. that can be used for detection In addition to the well, the target substance bound to the bead surface is labeled. By detecting fluorescence, color development, luminescence, magnetism and the like by this “labeling substance”, the presence / absence, the amount, etc. of the target substance can be measured, that is, “analysis of the target substance” can be performed. Further, after the washing in the previous operation, a “target substance extraction” can be performed by adding a solution capable of eluting the bound target substance and eluting the target substance. Furthermore, “purification of the target substance” can be performed by performing the same operation with the liquid containing the target substance to be purified.

Separation / detection / screening of cells: In “separation of cells”, for example, cells can be separated by placing the cells in a well by applying a liquid containing the cells onto the well plate. In the “cell detection”, cells can be detected by, for example, observing what has entered the well with a microscope or labeling with a labeling substance that specifically binds to the cells to be detected. Furthermore, in the “cell screening”, for example, by finding a specific binding of a pre-labeled antigen from various cells in a well, for example, B lymphocytes, based on the label. Cells can be screened.

<< Method for Producing Plate Member of the Present Invention >>
Next, the manufacturing method of the plate member of this invention is explained in full detail. The plate member of the present invention is
First, a “stamper” is manufactured from a “resist master” whose shape is imitated, and then injection molding is performed using the stamper as a mold (mold). Such a series of manufacturing steps will be described over time.

(Preparation of resist master)
Manufactures “resist master” used for mold production. First, a substrate that is a prototype of a “resist master” is prepared. The substrate to be prepared may be of any kind as long as its surface is flat and smooth. For example, it is possible to use a silicon wafer having a smooth surface that is polished in a mirror shape and having a thermal oxide film formed on the surface. Further, a substrate made of quartz or metal may be used.

  Next, a resist is formed on the surface of the substrate. After the resist is formed, a mask is disposed on the resist and exposure is performed. Exposure is not limited to collective exposure with a mask, but may be made by depiction in which exposure power is changed directly by a laser or electron beam. Furthermore, synchrotron radiation may be used as in the LIGA (Lithographie Galvanoformung Abformung) process.

  In the exposure, a multi-tone photomask that can freely change the tone of the mask may be used. As such a multi-tone photomask, a gray-tone mask and / or a halftone mask can be used. The gray tone mask makes a slit below the resolution of the exposure machine, and the slit part blocks a part of the light to realize intermediate exposure. On the other hand, the halftone mask realizes intermediate exposure by using a “semi-transmissive” film. In any case, three exposure levels of “exposed portion”, “intermediate exposed portion”, and “unexposed portion” can be realized by one exposure, and fine structures having different thicknesses after development can be obtained. In connection with the present invention, the height can be adjusted by adjusting the density of the gray tone mask and controlling the depth after development, so that the inclination angle of the side wall can be freely adjusted.

  After the exposure, development processing is performed. For example, development may be performed by immersing the exposed substrate in a TMAH (tetramethylammonium hydroxide) solution. By this development, a “master disk” simulating the shape of the microplate device is obtained (the obtained “master disk” can be referred to as “resist master disk” because it is obtained by patterning with a resist). .

(Production of stamper)
Next, a “stamper” is produced from the “resist master”. Specifically, the resist master is sputtered or plated to produce a stamper having a reverse pattern of the master pattern. For example, a stamper can be manufactured by forming a Ni sputtered film as a conductive film on the master and performing nickel sulfamate plating. The formation of the conductive film is not limited to being performed by sputtering, and may be performed using electroless plating, CVD (Chemical Vapor Deposition), or the like.

  As a plating solution used for the plating treatment, a nickel sulfamate plating solution having a low stress may be used, and pH buffering boric acid and Ni chloride that promotes dissolution of the anode Ni may be added thereto. Further, sulfamic acid may be used for pH adjustment of the plating solution so that the pH is always in the range of 3.8 to 4.2. The temperature of the plating solution may be 50 ° C., for example. Furthermore, the plating solution may always be subjected to filtration.

  The stamper material is not limited to Ni, but a metal such as Ag, Au, Bi, Cd, Co, Cr, Cu, Fe, In, Ni, Pd, Pt, Ru, Sn and / or Zn is used. May be.

(Injection molding of plate members)
Next, the plate member is manufactured by performing injection molding using the stamper as a mold. Specifically, the outer shape of the metal stamper is processed and attached to a molding machine, and then injection molding is performed. Various resins can be used as the molding resin. For example, a cycloolefin resin can be used. By such injection molding, the plate member of the present invention is finally obtained.

  As mentioned above, although embodiment of this invention has been described, it has only illustrated the typical example of the application scope of this invention. Therefore, those skilled in the art will readily understand that the present invention is not limited thereto and various modifications can be made.

● The number of steps on the side wall surface in the recess (the number of side walls) is not limited to two or three, but may be more than that. For example, it may be a side wall surface composed of 4 steps, 5 steps and 6 steps.

In electroforming used in the stamper manufacturing method, at least one selected from the group consisting of Ni, Ag, Au, Bi, Cd, Co, Cr, Cu, Fe, In, Pd, Pt, Ru, Sn and Zn However, the present invention is not necessarily limited to such an embodiment. For example, alloy plating mainly composed of one of Ag, Au, Bi, Cd, Co, Cr, Cu, Fe, In, Ni, Pd, Pt, Ru, Sn, and Zn, PTFE (polytetrafluoroethylene) ) And the like can be dispersed and incorporated into the plating film to produce a roll or flat plate stamper. In addition, Mn, Gd, Sm, W, Sb, Mo, P, B, S, etc. are made of a roll or flat plate alloy with increased hardness, lubricity and tenacity by actively incorporating them into the plating film. You can also make stampers.

  ● In this specification, the aspect of obtaining a resist master by wet plating using a plating solution has been mentioned, but it is not necessarily limited to such aspect, and dry plating such as hot dipping and vacuum plating (PVD, CVD, etc.) Even so, the effect of the present invention is not substantially changed.

  ● In this specification, the aspect of obtaining a resist master mainly by photolithography was mentioned. However, the present invention is not necessarily limited to this aspect, and any production method can be adopted as long as the same fine pattern and concave pattern can be formed. May be.

  ● In this specification, the aspect of obtaining the plate member mainly by injection molding was mentioned, but it is not necessarily limited to this aspect, and any molding method can be adopted as long as the same fine pattern and concave pattern can be formed. May be. For example, molding may be performed using UV curable resin or PDMS resin (polydimethylsiloxane), or a technique such as nanoimprint may be applied, or pattern transfer may be performed by hot embossing. Furthermore, optical modeling by laser light is also possible.

It should be noted that the present invention has the following aspects.

[First embodiment]: a fine pattern group in which a plurality of convex or concave fine patterns A having the same shape are formed,
In the cross section of the fine pattern A, a structure in which the maximum length portion Lmax gradually changes toward the shortest length portion Lmin is formed on a part or all of the side wall, and the surface is perpendicular to the central axis of the fine pattern A. An aggregate structure of fine patterns, characterized in that, when the cross-sectional area is measured over the entire length in the height direction H of the fine pattern A, the maximum length portion Lmax has a widest shape.
[Second Aspect]: In the first aspect, in the shape of the fine pattern A, the maximum length portion Lmax in the width direction of the fine pattern A has a length equal to or less than the pitch between the fine patterns A. object.
[Third Aspect]: In the first or second aspect, at least one side wall portion that is the cross-sectional shape of the fine pattern A and has an inclination angle with respect to the horizontal plane of 0 ° (excluding 0 °) to 90 °. A collective structure characterized by having more than one location.
[Fourth aspect]: In any one of the first to third aspects, when viewed as a cross-sectional shape of the fine pattern A, one or more corner portions of the fine pattern A are chamfered. Aggregate structure.
[Fifth Aspect]: In any one of the first to fourth aspects, when the cross-sectional shape of the fine pattern A is viewed, the corner portion of the fine pattern A is formed in a curved (arc) shape. A collective structure.
[Sixth aspect]: The aggregate structure according to any one of the first to fifth aspects, wherein the side wall portion is constituted by a chamfer and a curved portion when viewed as a cross-sectional shape of the fine pattern A. .
[Seventh aspect]: The aggregate structure of fine patterns is a substrate, and the substrate is cyclic polyolefin, cycloolefin resin, polycarbonate, amorphous polyolefin, polydimethylsiloxane, phenol resin, epoxy resin, melamine resin, urea resin, Saturated polyester resin, alkyd resin, polyurethane, polyimide, polyethylene, polypropylene, polyvinyl chloride, polystyrene, polyvinyl acetate, polytetrafluoroethylene, acrylonitrile resin, butadiene resin, styrene resin, acrylic resin, polyamide, polyacetal, modified polyphenylene ether, Polybutylene terephthalate, polyethylene terephthalate, polyphenylene sulfide, polysulfone, polyethersulfone, amorphous polyarylate, liquid crystal Rimmer, characterized in that at least one or more polymers selected from the group consisting of polyetheretherketone and Poamidoimido, the one set structure of the first to sixth aspect.
[Eighth Aspect]: The aggregate structure according to any one of the first to sixth aspects, wherein the fine pattern aggregate structure is a substrate, and the substrate includes silicon or glass.
[Ninth aspect]: The aggregate structure of fine patterns is a metal mold, and the metal mold is Ag, Au, Bi, Cd, Co, Cr, Cu, Fe, In, Ni, Pd, Pt. Any one of the first to sixth aspects, characterized in that it is a roll or a flat plate comprising at least one component selected from the group consisting of Ru, Sn and Zn Aggregate structure.
[Tenth aspect]: The fine pattern aggregate structure according to any one of the first to sixth aspects is a metal mold, and the metal mold is Mn, Gd, Sm, W, Sb, Mo. A metal mold characterized in that it is a roll or a flat plate comprising at least one component selected from the group consisting of P, B and S.

  In order to confirm the effect of the present invention, the following comparative examples and Examples 1 to 7 were carried out.

《Plate member production and bead collection rate confirmation test》
(Comparative Example 1)
As a well plate of the prior art, a plate member having a fine pattern having a concave shape as shown in FIG. 19 was produced. First, “Registry Master” was created. Specifically, hexamethyldisilazane (HMDS) is applied to “a silicon wafer substrate having a smooth surface and polished to a mirror surface, and a thermal oxide film formed on the surface”. Baked for a minute. Next, a positive photoresist for thick film (PMER P-LA900PM manufactured by Tokyo Ohka Kogyo Co., Ltd.) was applied at a thickness of 30 μm by spin coating. Subsequently, a chrome mask in which a fine well pattern is formed (see FIG. 20: a chrome mask in which holes with a diameter of 30 μm are regularly formed every 60 μm / actually used, the hole arrangement is a so-called “staggered arrangement”. The resulting chrome mask was placed on the resist, and contact exposure was performed with ultra-high pressure UV light. After exposure, the substrate was immersed in a 3% solution of TMAH (tetramethylammonium hydroxide) and developed to obtain a resist master. In FIG. 20, the black portion completely blocks light, and the white portion transmits UV light. Therefore, the white portion is recessed after exposure and development.

  After forming a Ni sputtered film as a conductive film on the obtained resist master, electroforming was performed. Specifically, nickel sulfamate plating solution was used, and pH buffered boric acid and Ni chloride for promoting the dissolution of anode Ni were added thereto. For adjusting the pH, sulfamic acid was used, and the pH was always adjusted to the range of 3.8 to 4.2. The temperature of the plating solution was 50 ° C., and the plating solution was always filtered. Electroforming was carried out under such conditions to obtain a stamper having a thickness of 400 μm.

  The outer shape of the obtained metal stamper was processed and attached to a molding machine, and then injection molding was performed using a cycloolefin resin as a molding resin (resin temperature: 340 ° C., mold temperature: 125 ° C., injection speed: 300 mm / s, holding pressure: 70 MPa). By such injection molding, a fine pattern plate member having a concave shape as shown in FIG. 19 was obtained.

Example 1
As Example 1 of the present invention, a “plate member provided with a fine pattern including concave portions as shown in FIG. 8” was prepared. That is, as shown in the drawing, a plate member having a fine pattern of recesses having a widest cross section of the opening was prepared.

  Specifically, hexamethyldisilazane was applied to “a silicon wafer substrate having a smooth surface and polished to a mirror surface, and a thermal oxide film formed on the surface”, and baked at 140 ° C. for 10 minutes. did. Next, a positive photoresist for thick film (PMER P-LA900PM manufactured by Tokyo Ohka Kogyo Co., Ltd.) was applied at a thickness of 30 μm by spin coating. Subsequently, a mask on which a fine well pattern was formed was placed on the resist, and contact exposure was performed with ultrahigh pressure UV light. In this example, a glass multi-tone mask having a light and shade as shown in FIG. 21 was used (a mask having a so-called “staggered arrangement of holes”). In FIG. 21, the black portion completely blocks the light, the white circle portion transmits the UV light, and the inclined gray portion transmits the UV light corresponding to the density of the shade. By controlling the transmittance in this way, the inclination angle of the recess side wall was adjusted. The pitch and arrangement of the individual patterns were set to 60 μm as in Comparative Example 1, and the well density was also set as in Comparative Example 1. Specifically, the wells in this example had Lmax shown in FIG. 8 of 60 μm and Lmin of 30 μm.

  After exposure, the substrate was immersed in a 3% solution of TMAH (tetramethylammonium hydroxide) and developed to obtain a resist master.

  After forming a Ni sputtered film as a conductive film on the obtained resist master, electroforming was performed. Specifically, nickel sulfamate plating solution was used, and pH buffered boric acid and Ni chloride for promoting the dissolution of anode Ni were added thereto. For adjusting the pH, sulfamic acid was used, and the pH was always adjusted to the range of 3.8 to 4.2. The temperature of the plating solution was 50 ° C., and the plating solution was always filtered. Electroforming was carried out under such conditions to obtain a stamper having a thickness of 400 μm.

  The outer shape of the obtained metal stamper was processed and attached to a molding machine, and then injection molding was performed using a cycloolefin resin as a molding resin (resin temperature: 340 ° C., mold temperature: 125 ° C., injection speed: 300 mm / s, holding pressure: 70 MPa). By such injection molding, a plate member having a fine pattern having a concave shape as shown in FIG. 8 could be obtained (an electron micrograph of the well formed in FIG. 22).

(Example 2)
As Example 2, a “plate member provided with a fine pattern including concave portions as shown in FIG. 4” was prepared. That is, as shown in the drawing, a plate member having a fine pattern of recesses having an opening having the widest cross section was prepared.

  The wells in Example 2 had Lmax shown in FIG. 4 of 60 μm and Lmin of 30 μm. In the first embodiment, the width of the opening is widened by one chamfering. However, in this second embodiment, the wells in which the inclination angles of the first side wall and the second side wall are changed by two chamfers as shown in FIG. Was made. The manufacturing method of the plate member itself is the same as in Example 1 (an electron micrograph of the well formed in FIG. 23 is shown).

(Example 3)
As Example 3, a “plate member provided with a fine pattern including concave portions as shown in FIG. 14” was prepared. That is, as shown in the drawing, a plate member having a fine pattern of recesses having an opening having the widest cross section was prepared. In Example 3, Lmax in FIG. 14 was 60 μm, and Lmin was 30 μm. The manufacturing method itself of the plate member is the same as that of the first embodiment.

Example 4
As Example 4, “a plate member provided with a fine pattern including concave portions as shown in FIG. 10” was prepared. That is, as shown in the drawing, a plate member having a fine pattern of recesses having an opening having the widest cross section was prepared. In Example 4, Lmax shown in FIG. 10 was set to 45 μm and Lmin was set to 30 μm (in the diagram schematically shown in FIG. Therefore, it is necessary to make Lmax smaller than the pitch width of the well). The plate member manufacturing method itself is the same as that in Example 1 (an electron micrograph of the well formed in FIGS. 24 to 27 is shown).

(Example 5)
As Example 5, “a plate member provided with a fine pattern including concave portions as shown in FIG. 12” was prepared. That is, as shown in the drawing, a plate member having a fine pattern of recesses having an opening having the widest cross section was prepared. In Example 5, Lmax in FIG. 12 was set to 60 μm and Lmin was set to 30 μm (unlike FIG. 10 of Example 4), Lmax can be expanded to the same width as the well pitch. The manufacturing method itself of the plate member is the same as that of the first embodiment.

(Example 6)
As Example 6, a “plate member provided with a fine pattern including concave portions as shown in FIG. 16” was prepared. That is, as shown in the drawing, a plate member having a fine pattern of recesses having an opening having the widest cross section was prepared. In Example 6, Lmax in FIG. 16 was 60 μm, and Lmin was 30 μm. The manufacturing method itself of the plate member is the same as that of the first embodiment.

(Example 7)
As Example 7, “a plate member provided with a fine pattern including concave portions as shown in FIG. 17” was prepared. That is, as shown in the drawing, a plate member having a fine pattern of recesses having an opening having the widest cross section was prepared. In Example 7, Lmax in FIG. 17 was set to 45 μm and Lmin was set to 30 μm (If Lmax is set to the same size as the pitch, the wells are in contact with each other and the rigidity is weakened. Therefore, it is necessary to make Lmax smaller than the well pitch width). The manufacturing method itself of the plate member is the same as that of the first embodiment.

(Bead collection efficiency confirmation test)
Using the plate member of Comparative Example 1 and Examples 1 to 7 (total of 10400 wells in length and breadth 320 in the center of the plate), the bead collection rate was examined.

  In the confirmation test, a substrate serving as a lid on which a flow path was separately prepared was pressure-bonded to a plate member. Over 110,000 zirconia beads were dispersed in water and the aqueous solution was allowed to flow. As a method of flowing beads, there are a method of passing only once and a method of reciprocating many times to pass the well part several times. In this confirmation test, the beads were passed only once. The beads that settled in the well were counted and this was repeated 10 times. The results are shown in Table 1.

  As can be seen from Table 1, compared to Comparative Example 1, Examples 1-7 generally had higher bead collection rates. That is, it was confirmed that the plate members of Examples 1 to 7 can effectively reduce the “well passage phenomenon”.

<Confirmation test of plate member stiffness characteristics>
In order to investigate the rigidity of the plate member of the present invention, structural simulation (comparison of well deformation by shape) was performed using CAE (Computer Aided Engineering).

  The well shape to be subjected to the structure simulation was the well shape of Comparative Example 1 (FIG. 19), Example 1 (FIG. 8), and Example 7 (FIG. 17). In order to set the collection rate to substantially the same value, the opening in Comparative Example 1 and the openings in Example 1 and Example 7 were set to the same diameter of 60 μm.

  FIG. 28 shows the well shape used in the simulation. The numerical value in the parenthesis in the height column is the division point of the shape. The well of Example 1 is tapered at a point of 15 μm from the bottom surface, and the well sidewall of the well of Example 7 changes at intervals of 10 μm.

The physical properties of the structure (plate member) were subjected to linear static analysis as a cycloolefin resin with a density of 1.01 g / cm ³ , a Poisson's ratio of 0.3, and a Young's modulus of 3100 MPa.

  In the analysis model, three wells were arranged vertically and horizontally, and the load was uniformly distributed only to the center well from one direction (stress was set to 15 MPa). The boundary condition of the bottom of the well was completely fixed, and the deformation and maximum displacement point under load were investigated. As a result, in the comparison with the same collection rate, when the maximum displacement of 0.439 μm in Comparative Example 1 is 100%, 36.9% in Example 1 is 0.162 μm, and 49.7% in Example 7. It was found that the thickness could be reduced to 0.218 μm. This is because, even if the opening of the well has the same diameter, the rigidity of the well is improved by making the cross section tapered, and it is difficult to deform with respect to the load.

  FIG. 29 shows a simulation result of the displacement amount of the well side wall when a load is applied. The horizontal axis indicates the distance from the bottom surface in the well cross section. In Comparative Example 1, since the well cross section is vertical, the distance from the bottom surface to the top surface is 30 μm, which is the same as the height. In the well shape of Example 1, since the taper shape is formed from the middle of the side wall, the length of the side wall cross section from the bottom surface to the top surface is longer than the height of the well. Furthermore, in the well of Example 7, the length from the bottom surface to the top surface is longer than the well height because the shape is complicated and the surface area is increased.

  As shown in FIG. 29, compared to the conventional well shape (Comparative Example 1), the well shape (Example 1 and Example 7) according to the present invention has about half the well deformation for the same load. I found out that As a result, when wells having the same opening diameter are arranged at the same density, it can be confirmed that the well shape in the present invention has a smaller amount of deformation for the same load and a structure with higher rigidity. It was.

  The plate member of the present invention can be used in the fields of medical science and bioscience. For example, when the plate member of the present invention is used, analysis, extraction, purification, cell separation, detection or screening of a target substance can be performed (particularly, such treatment / operation can be carried out in large quantities at once). Therefore, the plate member of the present invention can be used for various applications such as gene analysis, expression analysis, protein analysis, antigen / antibody reaction analysis, or cell analysis.

DESCRIPTION OF SYMBOLS 10 1st side wall part 20 2nd side wall part 30 3rd side wall part 40 Vertical side wall part 50 Recessed part (dent)
60 Body of plate member 70 Microbead 80 Liquid 100 Plate member

Claims (14)

A plate member having a fine pattern composed of a plurality of recesses of the same shape,
The side wall surface forming the concave portion is at least composed of a first side wall portion located on the opening surface side of the concave portion and a second side wall portion located on the bottom surface side of the concave portion,
The inclination angle α1 of the first side wall portion with respect to the horizontal plane (inclination angle on the solid portion side of the plate member) is the inclination angle α2 of the second side wall portion with respect to the horizontal plane (inclination angle on the solid portion side of the plate member). A plate member that is smaller than the plate member. 2. The plate member according to claim 1, wherein an inclination angle α <b> 1 of the first side wall portion is smaller by 5 ° to 60 ° than an inclination angle α <b> 2 of the second side wall portion. 3. The plate member according to claim 1, wherein among the width dimensions of the concave portion, the width dimension L _opening located closest to the opening surface when viewed along the vertical direction is the largest. The plate member according to claim 3, wherein the width L _opening has a length equal to or less than a pitch of the plurality of recesses. The plate member according to any one of claims 1 to 4, wherein an inclination angle α1 of the first side wall portion is 30 ° to 70 °. The plate member according to claim 1, wherein an inclination angle α <b> 2 of the second side wall portion is 75 ° to 90 °. The plate member according to claim 1, wherein at least one of the first side wall and the second side wall has a curved shape. The plate member according to claim 1, wherein a boundary portion between the first side wall and the second side wall is angular. The plate member according to claim 8, wherein the first side wall portion has a curved surface shape, and the second side wall portion has a planar shape. The plate member according to claim 1, wherein a boundary portion between the first side wall portion and the second side wall portion is smooth. The concave portion has a third side wall portion located further on the bottom side than the second side wall portion,
The inclination angle α3 of the third side wall portion with respect to a horizontal plane (inclination angle on the solid portion side of the plate member) is larger than the inclination angle α1 and the inclination angle α2. The plate member in any one of -10. The plate member according to claim 1, wherein a bottom surface of the concave portion forms a conical surface or a hemispherical surface. The concave portion further comprises a vertical side wall portion located on the opening surface side further than the first side wall portion,
The plate member according to any one of claims 1 to 12, wherein an inclination angle of the vertical side wall portion with respect to a horizontal plane is 90 °.   The metal metal mold | die used in order to shape | mold the plate member in any one of Claims 1-13.