JP5271672B2 - Plate-shaped container, mold used for forming the same, and processing method using the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a plate-like container housing particulate substances in respective wells one by one while securing a liquid amount necessary for the respective wells. <P>SOLUTION: The plate-like container is constituted by forming a large number of recessed parts to a plate-like member so as to house both of a solid substance A to be housed and a liquid substance B to be housed in the recessed parts and characterized in that housing restricting parts, which house the substances A to be housed in the respective recessed parts one by one while housing the substances B to be housed in the respective recessed parts, are formed in the respective recessed parts. <P>COPYRIGHT: (C)2010,JPO&amp;INPIT

Description

  The present invention relates to a plate-like container, and more particularly to a “container in which a plurality of recesses are formed in a plate-like member” such as a well plate or a microarray. The present invention also relates to a method for analyzing, extracting or purifying a target substance using such a plate-like container, and a method for separating, detecting or screening cells using a plate-like container. Furthermore, the present invention relates to a metal mold used for forming a plate-like container.

  In the field of medical science and bioscience, “plate-like containers with a large number of microwells” suitable for various treatments for separating, immobilizing, analyzing, extracting, purifying or reacting substances or cells are often used. (For example, refer nonpatent literature 1).

  As such a plate-like container, for example, a chip having a fine structure such as a microchemical chip, a biochip, or a DNA (deoxyribonucleic acid) chip is generally used in addition to a well plate, a microtiter plate, and the like. Unlike a microfluidic device that uses a liquid flow, such a plate-like container has micro concave portions that function as a container for containing a liquid arranged in an array, so that it is also a microarray, a microarray plate, a microarray chip, or the like. be called. In well plates and microarrays, many small wells are provided to process and analyze a large amount of information at one time, but the reaction is examined with a small amount of limited liquid proportional to the volume of such wells. . Recently, there are also protein array chips on which proteins such as antibodies are immobilized.

  When using the plate-like container, “solid matter such as particles” and “liquid used for reaction” are provided to each well. For example, each of the wells is provided with “a particle on which a substance such as nucleic acid, protein, sugar chain or cell is immobilized” and “a liquid containing a target substance such as a base, nucleic acid, protein, antigen or sugar chain”. Various analysis, extraction, purification, and the like can be performed by binding, adsorbing, or reacting the substance immobilized on the particle and the target substance. Recently, a technique has been proposed in which nucleic acid-immobilized microbeads are provided to each well for nucleic acid hybridization and sequencing.

Here, in consideration of the relative relationship between the microbead and the microreaction container part such as a well, the capacity of the reaction container part has a certain limitation. When it is desired to increase the capacity of one reaction container section (that is, the capacity of each well) in a conventional micro reaction container section represented by a well or the like, this is done by simply increasing the width dimension or depth dimension of the well. It is possible. However, when the width or depth of the well is increased, when microbeads are supplied to a well together with a liquid (≈reaction solution), there is a possibility that two or more microbeads are stored in one well. Come out. That is, in one plate-like container, there is a possibility that only one microbead can be stored in a certain well, whereas another well can store two or three microbeads. As a result, the ratio between the microbeads and the liquid amount (≈reaction liquid amount) in each well is different for each well. This means that the results of culture, inspection and analysis in the wells are different for each well. In addition, when analyzing components carried on individual microbeads for each micro reaction container part, if two or more microbeads are accommodated in one reaction container part (ie, well), the inspection method itself Will not hold true. On the other hand, if the well width is reduced so that only one microbead is provided for each well, the space inside the well for containing the reaction solution is inevitably reduced. Therefore, there is a concern that the necessary amount of the reaction solution cannot be secured, or that the microbead does not easily enter the well because the well opening is small.
Product information of Sumitomo Bakelite Co., Ltd. (Product name: Multiplate for culture) [online], [October 20, 2008 search], Internet <URL: http://www.sumibe.co.jp/sumilon/plate .html>

  The present invention has been made in view of the above circumstances. That is, the subject of this invention is providing the plate-shaped container which can accommodate one granular material in each well, ensuring the liquid amount required in each well.

In order to solve the above-described problems, the present invention is a plate-like container in which a plurality of concave portions are formed in a plate-like member so that both a solid object A and a liquid object B are accommodated. There,
The accommodation limiting part for accommodating one to-be-contained object A in each recessed part is formed in each inside (inside) of a recessed part, accommodating the to-be-contained object B in each recessed part. A plate-like container is provided.

  The present invention secures an appropriate amount of the object B (that is, a liquid such as a reaction liquid) in each of the recesses, while the “accommodation restricting portion” provided in each of the recesses allows the object A (that is, , A solid granular material) is accommodated one by one. In a preferred aspect, the storage restriction portion extends from the peripheral portion of the recess so as to have a form integrated with at least a part of the peripheral portion of each recess. That is, the accommodation limiting portion has a shape of a “peninsula” that is connected to the side wall (inner wall) of the recess. Moreover, in another certain suitable aspect, the accommodation limitation part has the form spaced apart with respect to the peripheral part of each recessed part. In this case, the accommodation limiting portion has an independent “island shape” or “pillar (pillar)” shape that is not connected to the side wall (inner wall) of the recess.

  In this specification, the “accommodation restricting portion” is a recess provided so that a desired number (ie, one) of objects to be accommodated such as granular materials can be accommodated while allowing the liquid to be accommodated / flowed. The internal shape / internal structure is substantially meant. That is, the “restriction” in the present specification refers to a mode in which a predetermined number of granular materials are regulated so as to be accommodated in the recesses on the assumption that fluid is secured in the recesses.

  In the present specification, the “plate-shaped container” generally has “a plurality of“ indentations ”that are used in the medical science field or the bioscience field for processing such as separation, immobilization, analysis, extraction, purification or reaction. Means "device or member". Preferably, the plate-like container has a thickness of about 100 μm to 50 mm because it is “plate-like”. Examples of such plate-plate containers include well plates, microtiter plates, microarrays such as microchemical chips, biochips, and DNA chips, microarray plates, or microarray chips. Therefore, when the plate-like container is a well plate or a microarray, the “recess” corresponds to what is called “well” (in the plate-like container, each well is spatially separated. ). Here, the term “micro” in this specification means that the size of a structure such as a recess is in the order of microns to millimeters (1 μm to several tens of mm) or “a plate-like container provided with a plurality of recesses”. Is substantially in the order of millimeters to centimeters (about 1 mm to several tens of centimeters).

  The solid inclusion A is not particularly limited, but may include particulates such as microbeads (beads), cells, microorganisms, bacteria, pollen, spores, and other biologically relevant particles. . Here, the “microbead” means a substantially spherical particle having a diameter of about 10 nm to several mm made of a material such as an inorganic substance such as resin, metal, or metal oxide (for example, the microbead is Particles having a diameter of about 1 μm to 5 mm made of sepharose, agarose, synthetic polymer, silica, alumina, zirconium oxide, yttrium-added zirconium oxide or iron oxide). In addition, the liquid object B is not particularly limited, and examples thereof include water, a solvent, a buffer, a reaction solution, and a fluid such as a sample liquid.

  In the plate-like container according to the present invention, each of the recesses has a width dimension and a depth dimension that are larger than the diameter dimension D of the object A. Therefore, even if the object A is stored in the recess, the liquid can be stored in the recess to some extent. The internal space of the recess excluding the storage restriction portion and the storage space for the object A becomes the storage space for the object B. The volume Vb of this storage space is increased to some extent, 9 to 30 times larger than the minimum volume assumed in the concave portion that can accommodate one object A). Specifically, for a cylindrical space having a bottom surface diameter and height having the same length as the diameter D of the spherical object A, the “object B” and the “spherical object having the diameter D” are used. Assuming that the volume of the object B assumed when the object A is accommodated is the minimum accommodation volume Vb ′, the “accommodation space volume Vb of the object B in the present invention” is 9 of the minimum accommodation volume Vb ′. Double to 30 times.

  When the accommodation restriction part has a form integrated with at least a part of the peripheral part of the concave part, the plate-like container of the present invention is regarded as “a device configured with a part of the side wall of the concave part deformed”. Can do. Thus, when the accommodation limiting portion has a form integrated with the peripheral portion of the recess, it is preferable that the horizontal cross section of the accommodation limiting portion is substantially rectangular, square, trapezoidal or kamaboko. That is, it is preferable that the accommodation limiting portion has a form protruding like a “bank” from the peripheral portion of the recess. In addition, if the shape of an accommodation restriction | limiting part restrict | limits an accommodation thing, there will be no restriction | limiting in particular. For example, the horizontal section of the storage restriction portion may be polygonal, round, or elliptical, or the corners (corners) of the above “rectangular horizontal section” may be rounded. Good.

  Moreover, when the accommodation restriction part has a form separated from the peripheral part of the recess, the accommodation restriction part is preferably provided so as to at least partially surround the accommodation space of the object A. That is, it is preferable that the accommodation restriction portions are aligned along the circumferential direction (circumferential direction when the center of the recess is the center of the circle). Further, when the accommodation limiting portion is separated from the peripheral portion of the concave portion, the concave portion is formed so that the dimension of the gap formed between the storage limiting portion and the peripheral portion of the concave portion is along the spiral direction. It is preferable that the width gradually increases from the peripheral portion toward the center. That is, it is preferable that the width of the space portion or the groove portion in the recess gradually increases as it approaches the center of the recess. Thereby, the to-be-contained object A will be easily guide | induced to the center part of a recessed part, and the reliable accommodation of the to-be-contained object A is accelerated | stimulated.

  Regardless of whether the accommodation restricting portion is integrated with the peripheral portion of the concave portion or separated from the peripheral portion, the accommodation of the object A in the concave portion is promoted. As such, the accommodation restriction may have various forms. For example, the “non-recessed surface that does not form a recess in the plate-like member” and the “upper surface of the accommodation limiting portion” may be non-planar. In particular, it is preferable that the height of the upper surface of the accommodation limiting portion is lower than the “non-recessed surface”.

  Further, the height of the accommodation limiting portion may be gradually lowered from the peripheral portion of the concave portion toward the center so that the object A is easily accommodated in the concave portion. Alternatively, the upper surface of the accommodation limiting portion may form an inclined surface (for example, an inclined surface inclined along a certain direction) inclined with respect to the horizontal plane. That is, the upper surface or the top surface of the accommodation limiting portion may be formed in a taper shape.

  In a preferred aspect, 2 to 6 accommodation limiting portions are formed for each of the recesses. When a plurality of storage restriction portions are provided in each recess, the height of the storage restriction portion sequentially changes along the circumferential direction in each recess so as to facilitate the storage of the object A in the recess. It may be. That is, the height of each of the accommodation restriction portions may be different in each recess.

  In addition, it is preferable that a recessed part and / or a non-recessed surface have hydrophilicity so that the liquid to-be-contained object B may flow into a recessed part easily. That is, it is preferable to apply “hydrophilicity” to the plate-like member. In order to impart hydrophilicity to the plate-like member, UV treatment, UV ozone treatment, plasma treatment, hydrophilic organic substance coating treatment, or metal thin film coating treatment may be performed. Such hydrophilization treatment is advantageous in that in the actual use of the plate-like container, bubbles are easily removed from the liquid B, or nucleic acids and proteins are difficult to adsorb to the plate-like member. It is.

  In a preferable aspect, in consideration of convenience in actual use of the plate-like container, a metal coating is provided only on the concave surface or on the concave surface and the surrounding non-concave surface. When the plate-like container is made of a transparent or semi-transparent material and a metal film is formed on the concave surface and / or non-recessed surface, the adjacent concave portions are exposed when light is emitted, fluorescent, or colored. The influence can be suppressed as much as possible. More specifically, for example, when a metal film is provided on the concave surface (that is, the well region), the transmittance of the measurement wavelength light in the wall between adjacent wells can be reduced. It is possible to prevent the measured values of the intensity of fluorescence and color from being influenced by the presence or absence of light emission or the intensity of adjacent wells. From the viewpoint of “reducing the transmittance of the measurement wavelength light between adjacent wells”, the present invention is not limited to an embodiment in which a metal coating is provided, and a light-absorbing dye, pigment, or the like is used for the material of the plate-like member (eg, resin ) May be mixed. However, even if the above-described metal coating is thin, the effect of reducing the light transmittance is particularly high. Therefore, it can be said that it is most effective to provide a metal coating in this respect. The metal used for the metal coating is, for example, selected from the group consisting of gold (Au), silver (Ag), copper (Cu), platinum (Pt), titanium (Ti), nickel (Ni) and alloys thereof. And at least one metal. Moreover, as a method using such a metal film, sputtering, vapor deposition, CVD, electroless plating, etc. can be mentioned. The thickness of the metal coating is preferably about 50 nm to 150 nm.

In the present invention, there are provided a metal mold used for forming the above plate-shaped container, a mold made of silicon or a compound thereof, or a resin mold. Such a mold is preferably in the form of a roll or a flat plate. Metal molds (or molds) are silver (Ag), gold (Au), bismuth (Bi), cadmium (Cd), cobalt (Co), chromium (Cr), copper (Cu), iron (Fe), It comprises at least one element selected from the group consisting of indium (In), nickel (Ni), palladium (Pd), platinum (Pt), ruthenium (Ru), tin (Sn) and zinc (Zn). It is preferable. In order to further improve the hardness, lubricity, tenacity, etc., the metal mold of the present invention includes manganese (Mn), gadolinium (Gd), samarium (Sm), tungsten (W), antimony (Sb). At least one element selected from the group consisting of molybdenum (Mo), phosphorus (P), boron (B) and sulfur (S) may be particularly positively included. Silicon or a compound template thereof is preferably silicon alone (Si), quartz (SiO ₂ ), silicon carbide (SiC), or the like. As the resin mold material, an acrylic resin, an epoxy resin, or the like can be used.

  Furthermore, in this invention, the various processing methods using the above-mentioned plate-shaped container are provided. Specifically, “beads capable of binding a target substance” to wells (ie, recesses) provided in a plate-like container (for example, “beads on which a substance / functional group capable of binding to a target substance is immobilized) )) One by one and a method for analyzing, extracting or purifying the target substance in each of the wells is provided. Also provided is a method of accommodating cells one by one in the wells provided in the plate-shaped container described above, and performing cell separation, detection or screening.

  In the plate-like container of the present invention, a predetermined number (that is, one) of granular materials can be accommodated in each recess while accommodating a necessary amount of liquid in each recess (≈well). Specifically, when the plate-shaped container of the present invention is used, a dispersion containing “a granular material composed of a plurality of objects to be stored A” and “a liquid of the object to be stored B” is removed from the plate-shaped container (in particular, a plurality of objects). In this case, one granular material can be accommodated in each well by the function of the “accommodation restricting portion”. In particular, the present invention is advantageous in that each well contains an appropriate amount of the liquid container B even though one granular material of the object A is stored in each well. In other words, in order to accommodate one granular material in the well of the prior art, the size of the well has to be reduced and the amount of liquid accommodated is inevitably reduced. It can be said that a particularly advantageous effect is achieved in that the amount of liquid accommodated is not reduced unnecessarily while the components are accommodated one by one. In a particularly preferred embodiment, the liquid contents B can freely travel through the entire inner region of each well.

  The plate-like container of the present invention restricts the granular material to be accommodated in the recess one by one by the action of the accommodation restricting portion. In order to prevent such an accommodation restriction part from acting excessively (that is, in order to prevent the inconvenience that the granular material does not enter the recess), the shape of the accommodation restriction part may lead the granular material to the center of the recess. It can be done.

  Further, in the present invention, by appropriately adopting a suitable form for each of the concave portion and the accommodation limiting portion, the liquid accommodation amount can be freely changed according to the application. In other words, in the present invention, the liquid volume of the well is used for the purpose of use while controlling so that only a desired number of particles (microbeads and cells) are contained in the well which is a fine reaction vessel. The desired capacity can be obtained accordingly.

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

<< General features of the present invention >>
As shown in FIG. 1, the plate-like container 100 of the present invention is formed by forming a plurality of recesses 30 in a plate-like member 50, and an accommodation restricting portion 25 is formed inside each recess. The accommodation restricting portion 25 functions to accommodate one object A in each recess 30 while accommodating the object B in each recess 30. The plate-like container 100 of the present invention can be designed freely according to the purpose of use with respect to the capacity of a fine well, and only the desired number of objects to be contained A (for example, “beads”) is provided in the well. It is controlled not to enter. Such a plate-like container 100 can be used for applications in the field of bioscience in which various reactions are performed in each recess, etc., so that “a reaction or the like in a small amount or a small amount of a solution formed on a plate” is performed. It can be regarded as a “reaction vessel assembly for performing in parallel”.

  The material of the plate-like container 100 of the present invention (that is, the material of the plate-like member 50) is not particularly limited, but cycloolefin resin (norbornene resin), cycloolefin copolymer resin, polycarbonate, amorphous polyolefin, Polydimethylsiloxane, phenol resin, epoxy resin, melamine resin, urea resin, unsaturated 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, polyphenylenesulfur De, polysulfone, polyether sulfone, amorphous polyarylate, liquid crystal polymer, may be at least one or more polymers selected from the group consisting of polyetheretherketone and Poamidoimido. The material of the plate-like container 100 of the present invention may be silicon or glass. Although the dimensions of the plate-like container of the present invention may vary depending on the application, for example, the lengths (L, H, W) as shown in FIG. 1 are L = 5 to 300 mm, H = 0. It may be about 2-50 mm and W = 10-300 mm. It is preferable that the width dimension and the depth dimension of the recess have dimensions larger than the diameter dimension D of the object A to be accommodated, so that an appropriate amount of the reaction liquid used for the reaction to be performed later can be accommodated in each well. It is like that. For example, the width dimension w of the recess may be about 0.05 μm to 20 mm, and the depth dimension h of the recess may be about 0.01 μm to 10 mm (see FIG. 1). In addition, although it can be said that it is preferable in many cases because the amount of liquid required for reaction increases if the width dimension and depth dimension of the recess are increased, a part in which a necessary component reacts due to an increase in moving distance. Therefore, unnecessary liquid cost is required due to the fact that the amount of necessary components exceeds the amount necessary and sufficient for the reaction or the like. For this reason, although it depends on the form of the recess and the accommodation restricting portion, the upper limit of the width dimension and the depth dimension of the recess is preferably about 10D or less, more preferably about 5D or less (D: of the object A to be stored) Average size). 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 recesses provided in the plate-like container 100 of the present invention varies depending on the application. For example, it is preferably 2500 to 100 million, more preferably 10,000 to 50 million, More preferably, it can be 50,000 to 10 million. In addition, the shape of the opening part of a recessed part does not have a restriction | limiting in particular, Circular, an ellipse, a polygon (for example, square or hexagon) etc. may be sufficient.

  In use, the plate-like container 100 of the present invention is used together with a dispersion containing “a granular material composed of a plurality of objects A” and “a liquid of the object B”. Specifically, the object A and the object to be accommodated in each of the recesses are provided by providing a dispersion liquid composed of the object A and the object B to the surface of the plate-like container where the recess is provided. The object B is accommodated together. At the time of such accommodation, although the sedimentation action resulting from the weights of the objects A and B may be used, they may be shaken or subjected to a centrifugal treatment as necessary. . Here, as described above, the object A may be a granular material such as a microbead (bead), a cell, a microorganism, a bacterium, pollen, a spore, and other biologically related particles (hereinafter, The object A is simply referred to as “particle” or “granular material”). Such a granular material may have various shapes such as a spherical shape, an elliptical shape, a rice granular 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”, as the name implies, means a shape like “rice grain” and generally refers to a shape in which the length of the particles is uniform in all directions, such as a spherical shape. In particular, it refers to a shape having no size anisotropy as a whole. From the viewpoint of facilitating accommodation in the recess, the object A is preferably spherical. The average size D of the objects A having a particle shape is, for example, preferably about 0.01 μm to 10 mm, and more preferably about 0.1 μm to 5 mm. Here, the “particle size” substantially means the maximum length among the lengths of the particles in all directions. The “average particle size” as used in the present specification refers to, for example, measuring the size of 300 particles based on a transmission electron micrograph or an optical micrograph of particles, and calculating the particle size calculated as the number average. Meaning. On the other hand, the object B is a fluid such as water, a solvent, a buffer, a reaction solution, and a sample liquid as described above (hereinafter, the object B is simply referred to as “liquid” or “reaction liquid”. Also called). There is no restriction | limiting in particular in the viscosity of the to-be-contained object B, and the to-be-contained object B should just have fluidity on the conditions when utilizing the component in the to-be-contained object B. FIG. The content of the object to be contained A of the dispersion liquid (that is, the dispersion liquid composed of the object to be contained A and the object to be contained B) supplied to the plate-shaped container is not particularly limited. About 50% by volume. The absolute amount or the number of the objects to be accommodated A is preferably such that all the objects to be accommodated A are accommodated one by one in the recesses, for example, the number of the recesses is 10,000. For example, the number of the objects to be stored A is preferably at least 10,000, and the object to be stored A having an absolute amount corresponding to the number is only required to be included in the dispersion.

  The accommodation restriction part 25 is provided inside each recess. In particular, the accommodation limiting portion 25 is provided so as to occupy a part of the internal space of the recess. Due to the accommodation limiting portion 25, “the accommodation space for the solid object A (= the particle accommodation space)” and “the accommodation space for the liquid object B (= accommodation of the liquid / reaction liquid) in each recess Space) ”is formed. Although the “accommodating space for the solid object A” has a size or shape that can accommodate one object A (particle), it can accommodate two or more objects A (particles). Has an unacceptable size or shape. In other words, it can be said that the accommodation restricting portion 25 partially restricts the internal space of the recess so that one granular material is accommodated. Thus, since it can be considered that the number of granular materials accommodated in the accommodation limiting portion 25 is limited, the “accommodation limiting portion” in the present invention can also be referred to as a “number limiting member”.

  For example, as shown in FIGS. 2A and 2B, the accommodation limiting portion 25 has a form integrated with at least a part of the peripheral portion of the recess, and extends so as to protrude from the peripheral portion of the recess. Exist. Further, for example, as shown in FIGS. 3 (a) and 3 (b) or FIGS. 5 (a) and 5 (b), the accommodation limiting portion 25 may have a form separated from the peripheral portion of the recess. Good.

When the accommodation restricting portion 25 extends from the peripheral portion of the recess, the accommodation restricting portion 25 is formed in the form of a “peninsula” connected to the side wall (inner wall) of the recess and the land as shown in FIG. have. In other words, it can be said that the concave portion has a wall-like structure in which an integral peninsula protrudes from the peripheral portion while avoiding the central portion.

  As shown in FIGS. 2A and 2B, an accommodation space 30 b for an object B (liquid) is formed around an accommodation space 30 a for an object A (particle). When a dispersion liquid composed of particles and liquid is supplied to the plate-like container, the particles are stored inside the recess due to gravity settling or the like. However, the storage limiting portion 25 exists inside the recess. Therefore, the particles are guided into the recesses so as to avoid it. As the particles enter the recesses, the liquid in which the particles are dispersed also enters the recesses. As a result, the interior of the recesses other than the particle storage space is filled with the liquid. As can be seen from the embodiment shown in FIG. 2B, a relatively large amount of liquid is accommodated in each recess, and preferably the liquid can flow relatively freely in each recess. (From this point of view, the liquid storage space 30b can also be referred to as a “flow path”). As described above, in the plate-like container of the present invention, an appropriate amount of liquid used for a reaction to be performed later can be stored in each recess. In addition, from the viewpoint of securing a certain amount of liquid storage space 30b while limiting the number of particles to be stored, the storage limiting portion 25 has a rectangular parallelepiped shape as shown in FIG. Is preferred. That is, it is preferable that the cross section of the accommodation restricting portion 25 has a rectangular shape as shown in FIG.

  In the case where the accommodation restricting portion 25 has a form separated from the peripheral portion of the recess, the accommodation restricting portion 25 is independent of the side wall (inner wall) of the recess, for example, as shown in FIGS. May have the form of a “pillar”. Preferably, as shown in FIG. 3 (b), the accommodation limiting portion 25 is provided so as to at least partially surround the accommodation space 30a of the article A to be accommodated. In this case, the accommodation space 30b for the object B (liquid) is formed around the accommodation space 30a for the object A (particle). When a dispersion liquid composed of particles and liquid is supplied to the plate-like container, the particles are stored inside the recess due to gravity settling or the like. However, the storage limiting portion 25 exists inside the recess. Therefore, the particles are accommodated inside (see FIG. 4). As the particles enter the recess, the liquid in which the particles are dispersed also enters the recess. As a result, the interior of the recess other than the particle storage space is filled with the liquid. As can be understood from the embodiment shown in FIG. 3 (b), an appropriate amount of liquid is secured inside each recess, and preferably the liquid can flow relatively freely inside the recess. In other words, as shown in the drawing, the gap formed between the “pillar-shaped accommodation limiting portion” and the “periphery portion of the recess” is connected to the outer periphery, so that it is accommodated in the recess. The liquid can freely travel through the entire space in the recess. Compared with the “peninsula” -shaped accommodation limiting portion 25 shown in FIG. 2, the “pillar (pillar)”-shaped accommodation limiting portion 25 as shown in FIG. You can do more.

  The number of the accommodation limiting portions 25 in each concave portion is not particularly limited to “one”, and may be about 2 to 10, preferably about 2 to 8, more preferably 2 to 6. It may be a degree. When the number of the accommodating restriction portions 25 in each concave portion increases, a preferred form for accommodating particles can be adopted as appropriate, so that the concave portions can be freely designed according to the application (for example, the particles once accommodated jump out of the concave portions). Design that does not do is possible). In general, when the number of the storage restriction portions 25 increases, the liquid storage amount is reduced due to the volume occupied by the storage restriction portion 25, but by providing a “liquid storage space” separately ( For example, see the embodiment of FIG. 11 to be described later), and a decrease in the liquid storage amount can be avoided.

  When the accommodation limiting portion 25 has a form separated from the peripheral portion of the concave portion, the accommodation limiting portion 25 is independent from the side wall (inner wall) of the concave portion, for example, as shown in FIGS. It may have a form of “island” or “island”. In particular, it is preferable that the dimension of the gap C formed between the accommodation restriction portion and the peripheral portion of the recess gradually increases as it goes from the peripheral portion of the recess to the center along the spiral direction. As a result, the “groove” formed by the gap C has a seamless structure that is continuous from the peripheral portion of the recess toward the center, so that the object A (particles) is more efficiently guided to the center of the recess. it can. Even in this embodiment, in each recess, the accommodation space 30b for the object B (liquid) is formed around the accommodation space 30a for the object A (particle). Therefore, when the dispersion liquid composed of particles and liquid is supplied to the plate-like container, the particles can be accommodated from the outside to the center of the recess along the gap C that gradually increases toward the center. As the particles enter the recess, the liquid in which the particles are dispersed also enters the recess. As a result, the interior of the recess other than the particle storage space is filled with the liquid. As can be understood from the embodiment shown in FIG. 5 (b), an appropriate amount of liquid is secured inside the recess, and preferably the liquid can flow relatively freely in the recess. In other words, as shown in the drawing, the recess formed due to the structure in which the gap formed between the “island-shaped accommodation limiting portion” and the “periphery of the recess” is connected at the outer periphery. The liquid accommodated in the interior can freely travel through the entire space in the recess. In particular, the island-shaped accommodation limiting portion preferably has a tapered shape in which the width dimension is reduced upward in the vertical direction, so that the liquid in the recess is particularly easy to flow. Compared with the “peninsula-like” accommodation limiting portion 25 shown in FIG. 2, the “island-like” or “isolated island-like” accommodation limiting portion 25 as shown in FIG. Can be more.

  In addition, when the accommodation restricting portion 25 has a form separated from the peripheral portion of the recess, the “non-recessed surface that does not form the recess in the plate-like member” and the “upper surface of the accommodation restricting portion” are not flush with each other. It may be. In particular, it is preferable that the height of the upper surface of the accommodation limiting portion is lower than the “non-recessed surface”. Thereby, the effect that accommodation of particle | grains is accelerated | stimulated may be show | played. For example, it is preferable that the upper surface of the accommodation limiting portion is lower than the “non-recessed surface” by about 0 (not including 0) to about 0.4 D μm, more preferably about 0.05 D to about 0.3 D (however, D is the diameter of the object A).

<< Various aspects of the accommodation restriction part >>
Various other forms of the “accommodation restricting portion” are conceivable. This will be described in detail below.

(Form with two peninsulas)
A mode in which two accommodation limiting portions are formed in each recess is shown in FIGS. 6 (a) and 6 (b) and FIGS. 7 (a) and 7 (b). Such an embodiment is an embodiment in which two so-called “peninsulas” are formed. That is, the two accommodation limiting portions extend from the peripheral portion of the recess toward the center. In particular, as shown in the drawing, two accommodation limiting portions are provided so as to have a symmetrical positional relationship with each other. In the embodiment shown in FIGS. 6A and 6B, the extending direction of the accommodation limiting portion 25 is shifted from the radial direction, and the particles are captured by the two accommodation limiting portions and the peripheral edge of the recess. Become. In the embodiment shown in FIGS. 7A and 7B, the two accommodation restricting portions 25 extend along the radial direction of the concave portion, and particles are captured from both sides by the two accommodation restricting portions.

  In the embodiment shown in FIGS. 6 (a) and 6 (b), the contained particles are captured at three points (e point, f point, and g point), so that the movement of the particles inside the recess is effective. Therefore, an advantageous effect can be obtained in that the contained particles are not easily ejected from the recess. Further, in the embodiment shown in FIGS. 7A and 7B, the particles are evenly captured from both sides by the accommodation restricting portion 25, so that the particles can be accommodated at the center of the recess, and as a result, the particle surface. An advantageous effect can be obtained in that the reaction between the particles and the liquid can be performed equally at any of the above points.

(Form with 5 peninsulas)
A mode in which five accommodation limiting portions are formed in each recess is shown in FIGS. Such an embodiment is an embodiment in which five so-called “peninsulas” are formed. That is, each of the five accommodation restriction portions extends from the peripheral portion of the recess toward the center. In particular, as shown in the drawing, five accommodation limiting portions 25 are provided along the radial direction. As can be seen from the illustrated embodiment, particles are captured at five points with five accommodation restriction portions. Since it is captured at five points, it is possible to effectively prevent the movement of the particles inside the recess, and as a result, an advantageous effect can be obtained in that the contained particles are not easily ejected from the recess.

(Form with 6 peninsulas)
9A and 9B show an embodiment in which six accommodation restriction portions are formed in each recess. In this embodiment, six so-called “peninsulas” are formed. That is, the six accommodation limiting portions extend from the peripheral portion of the recess toward the center. In particular, as shown in the drawing, six accommodation limiting portions are provided along the radial direction. As can be seen from the illustrated embodiment, particles are captured at six points with six accommodation restriction portions. Since it is captured at 6 points, it is possible to effectively prevent the movement of the particles inside the recess, and as a result, an advantageous effect can be obtained in that the contained particles are not easily ejected from the recess. In addition, as an aspect in which six accommodation restricting portions are formed in each recess, an aspect as shown in FIGS. 10A and 10B is also conceivable. This mode corresponds to the “accommodation restricting portion” and the “liquid accommodating portion” in FIG. 9 that are interchanged with each other, and the functions and effects brought about are the same as those in FIG. Further, as will be described later, by providing the upper surfaces of the six accommodation restriction portions so as to be inclined along the circumferential direction and / or the central direction, the particles can be easily guided into the recesses during the accommodation. Can do.

(Equipped with a liquid reservoir)
A mode in which a “liquid reservoir” is provided adjacent to the accommodation restriction portion is shown in FIGS. In such an embodiment, the portion corresponding to the “peninsula” is an area indicated by reference numeral 25a in the drawing, and the “liquid reservoir” is an area indicated by reference numeral 30b ′ in the drawing. In such an embodiment, a relatively large amount of liquid can be stored around the stored particles. Therefore, it can be said that the aspect in which the “liquid reservoir” is provided adjacent to the accommodation restriction part is an aspect particularly suitable for an application that requires a large amount of reaction liquid per particle. In this embodiment, in addition to the main reaction vessel portion (30a portion) in which particles are accommodated, a secondary reaction vessel portion (30b 'portion) formed by a "peninsula" extending from the side wall of the recess is provided. It can also be considered as an aspect.

(Form with peninsula inclined in the central direction)
FIG. 12 shows an aspect in which the “accommodation limiting portion inclined toward the center of the recess” is provided. The illustrated embodiment is an embodiment in which six “peninsulas” are formed. In particular, each accommodation limiting portion 25 has a tapered surface that decreases toward the center of the recess. That is, the height of the accommodation restriction portion is gradually lowered from the peripheral portion of the recess toward the center. In such an embodiment, when the particles are provided in the peripheral region of the recess, the particles can move so as to slide down the upper surface of the storage restriction portion due to gravity sedimentation or the like, so that the particles are easily stored inside the recess. That is, in the embodiment having the accommodation restriction portion as shown in FIG. 12, it is expected that the particle accommodation rate is improved. The inclination angle of the accommodation limiting portion (upper surface) with respect to the horizontal plane is preferably about 10 ° or more, more preferably about 20 ° or more, and further preferably about 30 ° or more (in addition, at an inclination angle of 0 °, The storage restriction part is not inclined, and the upper surface of the storage restriction part is horizontal). The upper limit of such an inclination angle is not particularly limited, but is appropriately selected from the relationship of the peripheral shape of the recess, the depth of the recess, and the like as long as the accommodation limiting portion at the center of the recess can function (the upper limit is It can be a value not exceeding 90 °). For example, the upper limit value of the tilt angle is preferably about 45 ° from the viewpoint of holding the particles once accommodated so as not to jump out of the recess.

(Form with a peninsula inclined in the circumferential direction)
FIG. 13 shows an aspect in which an accommodation limiting portion that is inclined along the circumferential direction of the recess is provided. The illustrated embodiment is an embodiment in which six “peninsulas” are formed, and in particular, each accommodation limiting portion 25 is along the circumferential direction of the recess (circumferential direction when the center of the recess is the center). It has a taper surface that is lowered. In other words, the upper surface 25A of the accommodation restricting portion is characterized in that it forms an inclined surface inclined in a certain direction with respect to the horizontal plane. In such an embodiment, when the particles are provided in the peripheral region of the recess, the particles can move so as to slide down the upper surface of the storage restriction portion due to gravity sedimentation or the like, so that the particles are easily stored inside the recess. That is, in the embodiment having the accommodation restriction portion as shown in FIG. 13, it is expected that the particle accommodation rate is improved. The angle of the inclined surface with respect to the horizontal plane is preferably about 10 ° or more, more preferably about 20 ° or more, and further preferably about 30 ° or more (note that the accommodation restriction portion is inclined at an inclination angle of 0 °). It indicates a state in which the upper surface of the accommodation restriction portion is horizontal). The upper limit value of the angle of the inclined surface is not particularly limited as in the above embodiment, but is appropriately selected from the relationship between the peripheral shape of the recess, the depth of the recess, and the like as long as the accommodation limiting portion at the center of the recess can function. (The upper limit value can be a numerical value not exceeding 90 °). For example, the upper limit value of the angle of the inclined surface is preferably about 45 ° from the viewpoint of holding the particles once accommodated so as not to jump out of the recess.

(Form with stepped peninsula)
FIG. 14 shows an aspect in which the heights of the plurality of storage restriction portions are stepwise different in the recess. The illustrated embodiment is an embodiment in which six “peninsulas” are formed, and in particular, the heights of the plurality of accommodation limiting portions 25 provided along the circumferential direction are sequentially changed stepwise. ing. That is, as shown in the drawing, the height of each of the accommodation limiting portions is different in each recess so as to sequentially change along the circumferential direction. In such an embodiment, when the particles are provided to the peripheral region of the recess, the particles can move so as to slide down the “upper surface of the stepped accommodation restriction portion” due to gravity sedimentation or the like. Easy to be accommodated. That is, in the embodiment having the accommodation limiting portion as shown in FIG. 14, it is expected that the particle accommodation rate is improved. The lower limit of the height that changes sequentially in stages (that is, the difference between “the height of a certain storage restriction portion” and “the height of an adjacent storage restriction portion”) is the diameter dimension D of the object A (particles). In general, 0.05D or more is preferable. When the difference in height is 0.05 D or less, it becomes difficult for the particles to move sequentially along the step. On the other hand, the upper limit of the height that changes in a stepwise manner (that is, the difference between “the height of a certain storage restriction part” and “the height of an adjacent storage restriction part”) is limited by the number n of the storage restriction parts. The value is D / n. When the height that changes stepwise sequentially becomes D / n or more, the height of the lowest part of the accommodation restriction portion that changes stepwise becomes zero, and the function of the accommodation restriction portion is reduced. It wo n’t be easy.

(Form with spiral peninsula)
A mode in which the spiral-shaped accommodation limiting portion is provided is shown in FIGS. 15 (a) and 15 (b). In the illustrated embodiment, the accommodation restriction portion 25 is provided in a spiral shape, and the height of the accommodation restriction portion is gradually lowered from the peripheral portion toward the center. Thus, the form of the accommodation limiting portion is not necessarily limited to an aspect extending linearly, and may be an aspect extending curvedly. In the embodiment shown in FIGS. 15A and 15B, when the particles are provided to the peripheral region of the recess, the particles move so as to slide down the “upper surface of the spiral-shaped accommodation restriction portion” due to gravity settling. Therefore, the particles are easily accommodated in the recesses. That is, in the embodiment of the concave portion having the accommodation limiting portion as shown in FIGS. 15A and 15B, it is expected that the particle accommodation rate is improved.

(Form with two pillars)
FIGS. 16A and 16B show an embodiment in which two “pillar-like (columnar)” accommodation restriction portions are formed in each recess. Such an aspect corresponds to a configuration in which one more notch portion P is provided with respect to the ring-shaped accommodation limiting portion as shown in FIG. As shown in the drawing, two accommodation restricting portions 25a and 25b are provided so as to have a symmetrical positional relationship with each other, and the two accommodation restricting portions surround "accommodated particles". In this mode, compared with the mode shown in FIGS. 3 (a) and 3 (b), the liquid stored around the particles can come and go more freely.

(Form with three pillars)
FIGS. 17A and 17B show an embodiment in which three “pillar-like (columnar)” accommodation restriction portions are formed in each recess. Such an embodiment corresponds to a configuration in which two more notch portions P are provided in the ring-shaped accommodation limiting portion as shown in FIG. As shown in the drawing, three accommodation restriction portions 25a, 25b, and 25c are provided so as to have a point-symmetrical positional relationship with each other, and the three accommodation restriction portions surround "accommodated particles". Yes. In this aspect, it can be said that the liquid accommodated around the particles can come and go more freely than in the aspect shown in FIG.

(Form with four pillars)
FIGS. 18A and 18B show an embodiment in which four “pillar-like (columnar)” accommodation restriction portions are formed in each recess. Such an embodiment corresponds to a configuration in which three more notch portions P are provided with respect to the ring-shaped accommodation limiting portion as shown in FIG. As shown in the figure, four accommodation restricting portions 25a, 25b, 25c, and 25d are provided so as to have a point-symmetrical positional relationship with each other, and the four accommodation restricting portions surround “accommodated particles”. It has become. In this mode, compared with the mode shown in FIG. 3, the liquid accommodated around the particles can come and go more freely.

(Equipped with pillars inclined toward the center)
FIG. 19 shows an aspect in which a “pillar-shaped accommodation limiting portion that is inclined toward the center of the concave portion” is provided. The illustrated embodiment is an embodiment in which four “pillars” are formed. In particular, each “pillar” has a tapered surface that decreases toward the center of the recess. That is, the height of the accommodation limiting portion 25 gradually decreases from the peripheral portion of the recess toward the center. In such an embodiment, when the particles are provided in the peripheral region of the recess, the particles can move so as to slide down the upper surface of the storage restriction portion due to gravity sedimentation or the like, and thus the particles are easily stored in the recess. That is, in the embodiment having the accommodation limiting portion as shown in FIG. 19, it is expected that the particle accommodation rate is improved. The inclination angle of the accommodation limiting portion (upper surface) with respect to the horizontal plane is preferably about 10 ° or more, more preferably about 20 ° or more, and further preferably about 30 ° or more (in addition, at an inclination angle of 0 °, The storage restriction part is not inclined, and the upper surface of the storage restriction part is horizontal). The upper limit of the inclination angle is not particularly limited as in the above embodiment, but is appropriately selected from the relationship between the peripheral shape of the recess, the depth of the recess, and the like as long as the accommodation limiting portion at the center of the recess can function. (Note that the upper limit value can be a numerical value not exceeding 90 °). For example, the upper limit value of the tilt angle is preferably about 45 ° from the viewpoint of holding the particles once accommodated so as not to jump out of the recess.

(Form with pillars inclined in the circumferential direction)
FIG. 20 shows an aspect in which a pillar-shaped accommodation limiting portion that is inclined along the circumferential direction of the recess is provided. The illustrated embodiment is an embodiment in which four “pillars” are formed. In particular, each “pillar” has a tapered surface that decreases along the circumferential direction of the recess. Yes. That is, the upper surface 25 </ b> A of the accommodation limiting portion forms an inclined surface that is inclined in a certain direction with respect to the horizontal plane. In such an aspect, when the particles are provided in the peripheral area of the recess, the particles can move so as to slide down the upper surface of the storage restriction portion due to gravity settling or the like, so that the particles are easily stored in the recess. ing. That is, in the embodiment having the accommodation restriction portion as shown in FIG. 20, it is expected that the particle accommodation rate is improved. The angle of the inclined surface with respect to the horizontal plane is preferably about 10 ° or more, more preferably about 20 ° or more, and further preferably about 30 ° or more (note that the accommodation restriction portion is inclined at an inclination angle of 0 °). It indicates a state in which the upper surface of the accommodation restriction portion is horizontal). The upper limit value of the angle of the inclined surface is not particularly limited as in the above embodiment, but is appropriately selected from the relationship between the peripheral shape of the recess, the depth of the recess, and the like as long as the accommodation limiting portion at the center of the recess can function. (The upper limit value can be a numerical value not exceeding 90 °). For example, the upper limit value of the angle of the inclined surface is preferably about 45 ° from the viewpoint of holding the particles once accommodated so as not to jump out of the recess.

(Form with stepped pillars)
FIG. 21 shows an aspect in which the heights of the plurality of pillar-shaped accommodation limiting portions differ in a stepped manner in the recess. The illustrated embodiment is an embodiment in which four “pillars” are formed. In particular, the heights of a plurality of “pillars” provided along the circumferential direction are sequentially changed stepwise. Yes. That is, as shown in the drawing, the height of each of the accommodation limiting portions is different in each recess so as to sequentially change along the circumferential direction. In such an embodiment, when the particles are provided to the peripheral region of the recess, the particles can move so as to slide down the “upper surface of the stepped accommodation restriction portion” due to gravity sedimentation or the like. Easy to be accommodated. That is, in the aspect having the accommodation restriction portion as shown in FIG. 21, it is expected that the particle accommodation rate is improved. The lower limit of the height that changes sequentially in stages (that is, the difference between “the height of a certain storage restriction portion” and “the height of an adjacent storage restriction portion”) is the diameter dimension D of the object A (particles). In general, 0.05D or more is preferable. When it becomes 0.05 D or less, it becomes difficult for the particles to move sequentially along the step. On the other hand, the upper limit of the height that changes in a stepwise manner (that is, the difference between “the height of a certain storage restriction part” and “the height of an adjacent storage restriction part”) is limited by the number n of the storage restriction parts. The value is D / n. When the height that changes stepwise sequentially becomes D / n or more, the height of the lowest part of the accommodation restriction portion that changes stepwise becomes zero, and the function of the accommodation restriction portion is reduced. It wo n’t be easy.

(Form with spiral pillars)
A mode in which a spiral “pillar” is provided is shown in FIGS. In the illustrated embodiment, the “pillar” is provided in a spiral shape, and the height of the accommodation limiting portion is gradually lowered from the peripheral portion toward the center. In such an embodiment, when the particles are provided to the peripheral region of the recess, the particles can move so as to slide down the “upper surface of the spiral-shaped accommodation restricting portion 25” due to gravity sedimentation or the like. Easy to be accommodated. That is, in the embodiment of the concave portion having the accommodation limiting portion as shown in FIG. 22, it is expected that the particle accommodation rate is improved.

(Form with cylindrical pillars)
An embodiment in which four cylindrical “pillars” are formed in each recess is shown in FIGS. Such an aspect corresponds to a configuration in which each of the four accommodation restricting portions illustrated in FIG. 18 has a cylindrical shape. As shown in the figure, four accommodation restriction portions 25a to 25d are provided so as to have a point-symmetrical positional relationship with each other, and the four accommodation restriction portions surround "accommodated particles". In this mode, it can be said that the liquid contained around the particles can come and go more freely than the mode shown in FIG.

(Configuration with “island-like” accommodation restriction section—modified example 1)
An embodiment in which three “island-like” accommodation restriction portions are formed in each recess is shown in FIGS. The accommodation limiting portion of this aspect has a shape separated from the peripheral portion of the recess as in the case of “pillar”, but does not have a geometrical shape and an angular shape as much as “pillar”. The whole surface is smooth. In such an aspect, the dimension of the gap C formed between the “accommodation restricting portion” and the “peripheral portion of the recess” gradually increases as it goes from the peripheral portion of the recess toward the center along the spiral direction. ing. For example, as illustrated, the dimension _G a ~G _f gap C, larger in the _{G b} from _{G a,} also towards the _{G c} is greater than _{G b} (hereinafter, the same). In such an embodiment, the provided particles can be guided from the outside to the center of the recess along the gap C that gradually increases toward the center. That is, since the particles are easily guided to the central portion of the concave portion, an advantageous effect can be obtained in which the accommodation of the particles is promoted.

(Configuration with “island-like” accommodation restriction section—Modification 2)
An embodiment in which four “island-like” accommodation restriction portions are formed in each recess is shown in FIGS. Also in this embodiment, as in the embodiment shown in FIG. 24, the dimension of the gap C formed between the “accommodation restricting portion” and the “peripheral portion of the recess” extends from the peripheral portion of the recess to the center so as to follow the spiral direction. And gradually getting wider. For example, as illustrated, the dimension _G a ~G _f gap C, larger in the _{G b} from _{G a,} also towards the _{G c} is greater than _{G b} (hereinafter, the same). Even in such an embodiment, the particles can be guided from the outside to the inside of the recess along the gap C that gradually increases toward the center. That is, since the particles are easily guided to the central portion of the concave portion, an advantageous effect can be obtained in which the accommodation of the particles is promoted.

(Configuration with “island-like” accommodation restriction section—Modification 3)
Other modified examples of the “island” accommodation restriction portion are shown in FIGS. The “island-shaped” accommodation limiting portion shown in the figure is the same as that shown in FIGS. 24 and 25 in the point of effect, but is different in the shape. Although the accommodation restriction part 25 shown in FIGS. 24 and 25 has a shape separated from the peripheral part of the recess to the last, the accommodation restriction part 25 shown in FIGS. It is integrated. Although the shape is different in this respect, even in the accommodation restricting portion shown in FIGS. 26 to 28, the concave portion is formed so that the dimension of the gap formed between the accommodation restricting portion and the peripheral portion of the concave portion is along the spiral direction. It should be noted that it gradually becomes wider as it goes from the peripheral part to the center. That is, the particles can be guided from the outside to the inside of the recess along the gap that gradually widens toward the center.

  In the case of the island-shaped accommodation limiting portion as shown in FIGS. 26 to 28, although the gap (that is, “groove”) formed in the recess is not connected at the outer peripheral portion, the gap widens upward. Due to the tapered shape, the movement of the solution in the well is relatively easy.

<Capacity of liquid container>
The plate-like container of the present invention is characterized in that “the capacity of the liquid container” is large as described above. This will be described in detail.

(Capacity of liquid container in the prior art) -Cylindrical well A cylindrical well 30 'as seen in the prior art is shown in FIGS. 29 (a) and 29 (b). When the diameter of the spherical particle 60 provided to the well is D, when the illustrated container diameter is 2D, two spherical particles are contained in the well. Therefore, if the diameter of the well is D or more and 2D or less, two spherical particles are not accommodated in the well. However, it must be taken into account here that the well is actually a three-dimensional shape. In a state where the volume of the spherical particles is accommodated in the well more than half, it is difficult for the spherical particles to come out of the container again (see FIG. 30). For this reason, if the state in which more than half of the second spherical particles are accommodated in the well is regarded as “impossible”, the diameter of the well must be 1.866 D or less. The basis for such calculation is shown below.

About the calculation basis of "1.866D or less"

Here, the smallest possible cylindrical well is a cylindrical well having a bottom diameter and a height having the same length as the diameter D of the spherical particle (that is, shown in FIGS. 29A and 29B). Well). The volume of such a cylindrical well is D ³ π / 4. In this case, if the volume D ³ π / 6 of the spherical particles (diameter D) to be stored is taken into consideration, the capacity of the liquid storage portion is the difference between the well volume and the spherical particle volume (D ³ π / 4). −D ³ π / 6). That is, it can be said that the minimum liquid storage capacity assumed when storing one spherical particle is (D ³ π / 4-D ³ π / 6) (hereinafter also referred to as “minimum liquid capacity”).

When the diameter of the cylindrical well as shown in FIGS. 29 (a) and 29 (b) is 1.866D calculated above (that is, assuming the maximum diameter to ensure that two spherical particles are not accommodated) In this case, the capacity of the liquid storage portion is “((1.866D) ² · D · π−D ³ π / 6)” (here, the depth of the recess is assumed to be D as usual). Assuming that the minimum volume (D ³ π / 4-D ³ π / 6) of the same depth is used, “((1.866D) ² · D · π−D ³ π / 6) / (D ³ π / 4-D ³ π / 6) ”, which is only about 8.4 times. This is because, in the conventional cylindrical well, in which only one spherical particle is contained while ensuring the liquid capacity, the liquid capacity has only a capacity of about 8.4 times the above minimum capacity. It means not to.

(Capacity of liquid container in the prior art) -square column shaped well A square column shaped well 30 '' as seen in the prior art is shown in FIGS. 31 (a) and 31 (b). When the diameter of the spherical particle 60 provided to the well is D, when the illustrated container diameter is 2D, two spherical particles are contained in the well. Therefore, if the length of one side of the well is not less than D and not more than 2D, two spherical particles are not accommodated in the well. However, as in the case of the “cylindrical well” described above, it must be considered that the well is actually a three-dimensional shape. In a state where the volume of the spherical particles is stored in the well more than half, it is difficult for the spherical particles to come out of the container again (see FIG. 32). For this reason, if the state in which more than half of the second spherical particles are accommodated in the well is regarded as “impossible”, the length of one side of the well must be 1.61 D or less with respect to the diameter D of the spherical particles. (The calculation process can be the same as the case of the above-mentioned “cylindrical well”, and is omitted to avoid duplication).

Here, the smallest possible quadrangular prism-shaped well is a quadrangular prism-shaped well having the same width and height as the diameter D of the spherical particle (that is, in FIGS. 31A and 31B). Well as shown). The volume of such a square pole shape well is D ^3. In this case, when the volume D ³ π / 6 of the spherical particles (diameter D) to be accommodated is taken into consideration, the capacity of the liquid storage portion is (D ³ -D ³ π / 6). That is, it can be said that the minimum capacity of the liquid storage portion assumed in the case of storing one spherical particle is (D ³ -D ³ π / 6).

When the length of one side of a quadrangular prism-shaped well as shown in FIGS. 31 (a) and 31 (b) is 1.61D calculated above (that is, it is guaranteed that two spherical particles are not accommodated). The volume of the liquid container is a cube of height D with a square having a side of (1.61D) as the bottom, that is, from (1.61D) ² · D to the amount of spherical particles The value is obtained by subtracting the volume “D ³ π / 6”. This is (2.5921D ³ -D ³ π / 6), and when divided by the minimum volume (D ³ -D ³ π / 6), the capacity is only about 4.3 times.
This is because, in the prior art square columnar well, in which only one spherical particle is contained while ensuring the liquid capacity, the liquid capacity is only about 4.3 times the above-mentioned minimum capacity. It means not having.

(Capacity of the liquid storage portion in the present invention)-well in the mode shown in Fig. 2 With respect to the size of the "storage limiter" in the mode shown in Figs. 2 (a) and 2 (b), the length dimension m is D and the width dimension When n is 0.2D (well diameter dimension: 2D), considering the volume D ³ π / 6 of the spherical particles (diameter D) to be accommodated, the capacity of the liquid container is equal to the volume of the well (D ³ The value obtained by subtracting the volume of the spherical particles (D ³ π / 6) and the volume of the accommodation restriction portion of about 0.2D ³ from π) (the volume of the arc portion of the accommodation restriction portion is not considered because it is minute). In other words, the capacity of the liquid container is divided by the above-mentioned minimum volume equation to obtain “((D ³ π) − (D ³ π / 6) − (0.2D ³ )) / ((D ³ π / 4− D ³ π / 6)) ”, and therefore, the capacity of the liquid container is 9.2 times the minimum volume (D ³ π / 4−D ³ π / 6).
Here, if the shape shown in FIG. 2 is D = 25 μm, and the depth of the recess is 25 μm,
Minimum volume ^{(D 3 π / 4-D} 3 π / 6) , the 12271.8-8181.2 = 4090.6
Well volume (D ³ π) = 49087.4
Volume of spherical particles (D ³ π / 6) = 8181.2
The volume of the accommodation restriction part is about (0.2D ³ ) = 3125
49087.4- (8181.2 + 3125) = 37781.2
Thus, 37781.2 ÷ 4090.6 = 9.2 times.

(Capacity of the liquid storage portion in the present invention)-well of the embodiment shown in Fig. 3 The dimensions of the "storage restriction portion" of the embodiment shown in Figs. 3 (a) and (b), the outer diameter 2D and the inner diameter 1.2D, When the width dimension k of the notch P is 0.4D (well diameter dimension: 3.6D), the volume D ³ π / 6 of spherical particles (diameter D) to be accommodated is calculated.
From ^{(3.6D) 2/4 · π} · D,: well volume
Accommodate limited outsider径体product: ^(2D) subtract ^2/4 · ^π · D,
Accommodating restricted inner diameter volume: ^(1.2D) plus ^2/4 · ^π · D,
The volume of the notch P≈0.4D · (2D−1.2D) / 2 · D is added,
The volume of the spherical particles (diameter D) to be accommodated is a value obtained by subtracting D ³ π / 6.
The capacity of the liquid storage portion is about 7.81D ³ , which is about 30 times the minimum volume (D ³ π / 4−D ³ π / 6) = 0.26D ³ .

(Capacity of the liquid storage portion in the present invention)-well of the embodiment shown in FIG. 5 With respect to the dimensions of the “storage restriction portion” of the embodiment shown in FIGS. When the dimension r = 0.5D and the height dimension s = D (well width dimension: 2.7D), considering the volume D ³ π / 6 of the spherical particles (diameter D) to be accommodated, the liquid container The capacity is about 16 times the minimum volume.
For example, when D is 25 μm and the depth of the recess is 25 μm,
Total volume of well ≒ 99500μm ³
Containment restriction part≈8200 μm ³ × 3 = 24600 μm ³
Volume D ³ π / 6≈8181.2 μm ^{3 of} spherical particles (diameter D) accommodated
Minimum volume (D ³ π / 4-D ³ π / 6) ≈4090.6 μm ³
So that
{(99500)-(24600)-(8181.2)} / 4090.6 = 16.3
It becomes about 16 times the minimum volume.

Hereinafter, similarly, for the modes shown in FIGS. 6, 8, 9, 16, 17, 18, 18, 24, 25, and 26, the minimum volume (D ³ π / 4-D ³ π / 6) is used. The capacity of the liquid container with respect to (= multiple of the minimum volume) is shown in the following table.
[Table 1]

Table 2 summarizes the “capacity (multiple) of the liquid storage portion” in the various storage restriction portions described above.
[Table 2]

<< Method for Manufacturing Plate-shaped Container of the Present Invention >>
Next, the manufacturing method of the plate-shaped container of this invention is explained in full detail. The plate-like container of the present invention is manufactured by first producing a “stamper” from what is called a “resist master” whose shape is imitated, and then performing injection molding using the stamper as a mold (mold). it can. 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. A substrate made of quartz 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. For example, a chrome mask in which holes having a pattern as shown in FIG. 33A are regularly provided (the mask shown in FIG. 33A is a mask used when forming “three island-like accommodation restriction portions”) is arranged. Then, contact exposure is performed. Exposure is not limited to batch exposure using a mask, and exposure may be performed directly using a laser or an electron beam. Furthermore, synchrotron radiation may be used as in the LIGA (Lithographie Galvanoformung Abformung) process.

  Note that in the case of forming an accommodation restriction portion having a height dimension change or an accommodation restriction portion having an inclined surface, that is, in the case of forming an accommodation restriction portion having a change in the depth direction in the well, the gradation of the mask. It is possible to use a multi-tone photomask that can freely change. 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.

  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” simulating the shape of the plate-like container is obtained (the obtained “master” is obtained by patterning with a resist, and can be referred to as “resist master”). .

(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. That is, the plating process may be performed using such a metal. Also, alloy plating or PTFE (polytetrafluoroethylene) mainly composed of any metal of Ag, Au, Bi, Cd, Co, Cr, Cu, Fe, In, Ni, Pd, Pt, Ru, Sn or Zn. Etc. can be dispersed and taken into the stamper material to produce a roll-shaped or flat plate metal mold. Furthermore, Mn, Gd, Sm, W, Sb, Mo, P, B, and / or S may be positively included as a stamper material in order to improve hardness, lubricity, and tenacity.

(Plate container injection molding)
Next, a plate-like container 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-like container of the present invention is finally obtained (FIG. 33 (b) shows the overall shape of the plate-like container. In particular, FIG. 33 (b) shows hatching. "Well formation region" can be understood). The molding resin is not limited to cycloolefin resin, but PC (polycarbonate resin), PP (polypropylene resin), polystyrene, poly (meth) acrylic acid, poly (meth) acrylic acid ester, polyvinyl ether, polyurethane, polyamide, Polyvinyl acetate, polyvinyl alcohol, polyallylamine, polyethyleneimine, and the like may be used. Furthermore, UV curable resin or PDMS resin (polydimethylsiloxane) may be used.

  The stamper pattern is not necessarily limited to injection molding, and the stamper pattern may be transferred by hot embossing. Further, the substrate may be made of glass such as Si or quartz, and the well may be directly produced by performing wet etching or dry etching after pattern exposure. Further, the well can be produced by performing sandblasting or fine machining instead of wet etching or dry etching. In injection molding, it is necessary to provide a so-called “draft angle” in the mold in consideration of the resistance at the time of mold release. However, in the case of directly producing a well, such “draft angle” is not considered. This is advantageous in that the accommodation limiting portion can be formed.

  Thus, the plate-like container of the present invention is preferably made of a resin layer made of resin or formed on a support, and as a result, mass production means such as injection molding and imprint can be utilized. Low-cost mass production is possible.

<< Various treatment methods using the plate-like container of the present invention >>
Next, various treatment methods using the plate-like container of the present invention will be described. When the plate-like container of the present invention is used, the target substance can be analyzed, extracted or purified, and various cells can be separated, detected or screened. These techniques are the same as those performed using conventional well plates in fields such as medical science and bioscience.

(Analysis, extraction and purification of target substances)
In “analysis of target substance”, for example, a specimen liquid containing or possibly containing a target substance or a mixture of a specimen liquid and another liquid (hereinafter referred to as a specimen-containing liquid) and an antigen that specifically binds to the target substance Then, a bead having an antibody, a nucleic acid, or the like immobilized on the surface is brought into contact in the well and conditions for sufficiently binding the target substance to the bead surface are given, and then the sample liquid is removed or washed away, and washing is repeated 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, the 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.

  As mentioned above, although embodiment of this invention has been described, this invention is not limited to this, Various modifications can be made.

  For example, the overall form of the plate-like container is not limited to the form shown in FIG. 1, and a form suitable for the application may be appropriately adopted. In addition to the well structure, the plate-like member in the plate-like container of the present invention may be provided with a structure such as a groove, a protrusion, and / or a flow path. If a specific example is given, the form of the plate-shaped container of this invention may have a form as shown to FIGS. 34 and 35, a dispersion liquid (for example, a dispersion liquid in which beads are dispersed in a reaction liquid) can be supplied from the outside of the well region. Further, in the form shown in FIG. 36, columnar protrusions are formed on the plate surface adjacent to each well, and when the dispersion liquid is supplied, the beads and the columnar protrusions may collide, so that the beads enter the wells. Easy to use.

Further, when a plurality of storage restriction portions are provided in each well, the storage restriction portions do not have to be the same size, and may be different from each other. For example, in the case of a pillar-shaped accommodation restriction portion, a large, medium, and small columnar accommodation restriction portion may be formed to adjust the liquid capacity in the well. Further, the form of the accommodation restriction portion is not limited to the illustrated embodiment, and may be freely changed as necessary (that is, the width, height, height, etc. of the accommodation restriction portion depend on the application. Freely selectable). For example, in the case of the pillar-shaped accommodation limiting portion, the shape can be freely selected, and the shape may be a rectangle, a triangle, or a complicated shape. That is, in addition to a conical storage restriction portion, a quadrangular pyramid shape, a triangular pyramid shape, a quadrangular prism shape, and a triangular prism shape accommodation restriction portion are also conceivable. In addition, the thickness of one pillar-shaped accommodation limiting portion may be changed gradually, or the thickness of one pillar-shaped storage restriction portion is changed stepwise in the middle. It may be. Furthermore, each surface of the accommodation limiting portion is not limited to a “planar shape”, and may be formed in a sharp or curved shape. For example, the upper surface of the peninsula-shaped or pillar-shaped accommodation limiting portion may be formed to be curved in a curved shape.

  Furthermore, in consideration of convenience in actual applications, other separate plates, holders, and / or “parts that perform heating, stirring, pumping, detection, etc.” for the plate-like member in the plate-like container of the present invention ” Etc. may be combined. In an actual application, the dispersion liquid may be used only for accommodating the particles (contained object A) in each recess, and then the reaction liquid or the like (contained object B) may be used separately. Specifically, after the particles are accommodated using a dispersion liquid in which particles are dispersed in an appropriate medium a, the medium a is removed or reduced from the recesses, and then the desired reaction liquid (contained material B). Etc. may be accommodated in the recess.

  While actually producing the plate-shaped container of the present invention, a test for confirming the effect of the plate-shaped container of the present invention was conducted.

<< Production of plate-like container >>
First, a “resist master” was created. 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 the surface was 10 ° C. at 140 ° C. 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 chromium mask on which a fine well pattern was formed was placed on the resist, and contact exposure was performed with ultrahigh pressure UV light. The UV light source used for this exposure included g-line (436 nm), h-line (405 nm) and I-line (365 nm). After the exposure, the substrate was immersed in a 3% solution of TMAH (tetramethylammonium hydroxide) and developed to obtain a “resist master” in which the shape of the plate-like container was imitated.

  After forming a Ni sputtered film as a conductive film on the resist master, a reversal pattern stamper (for example, having a thickness of 400 μm) was prepared by performing nickel sulfamate plating. Injection molding was performed by using the obtained stamper as a mold. Specifically, the outer shape of the 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 this injection molding, the plate-shaped container of the present invention was obtained (see FIG. 37).

  In the case of using the mask shown in FIG. 33A, a well plate having “three island-like accommodation restriction portions” as shown in FIG. 5 formed in each well could be manufactured. Such a well plate has a plate form (horizontal dimension: 75 mm, vertical dimension: 25 mm, thickness: 1 mm) as shown in FIG. 33 (b), and a well formation region (lateral dimension) at substantially the center of the plate surface. : 40 mm, vertical dimension: 16 mm). The number of wells formed was 100,000 (width dimension of each well: about 68 μm, well depth: about 26 μm).

<Effect confirmation test of hydrophilic treatment>
A test was conducted to confirm the effect of “hydrophilicity” that can be applied to the concave and / or non-concave surfaces. Specifically, UV ozone treatment was performed on the well plate as shown in FIG. 10 using a UV ozone treatment device (Iwasaki Electric Eye UV-ozone cleaning device OC-250616-DA). As a result, the contact angle with respect to water of the non-recessed surface where no well was formed decreased from 82 degrees before the treatment to 20 degrees. When an aqueous dispersion containing beads was placed on the well formation region, it was difficult to uniformly place the beads in the well because the water became droplets and the beads gathered around the well before treatment. However, one bead could be placed in each well substantially uniformly after the treatment. In addition, the removal of bubbles was improved after the treatment.

  In order to apply “hydrophilicity”, a metal thin film coating treatment may be performed in addition to the UV ozone treatment. In order to confirm whether or not such a coating treatment is possible, a metal made of Ti is formed on the surface of the well plate. It is also noted that the thin film (50 nm) was actually created by sputtering.

<Confirmation test of the storage effect of the plate-like container>
In order to confirm the effect of the accommodation restriction portion in the recess, it was examined how the particle accommodation rate changes due to the difference in the shape of the accommodation restriction portion (In Examples 1 to 6 below, the well plate The above-mentioned hydrophilic treatment was used).

Example 1
A test was conducted to confirm the effect of “a peninsula-shaped accommodation restriction portion whose height gradually decreases toward the center”. First, a “well plate having an accommodation restriction portion inclined in the central direction in each well” prepared by the above-described manufacturing method was prepared (see FIG. 12). Such a housing restriction portion whose height changes can be realized by using a halftone mask for exposure. In addition, 8 types of well plates were prepared so that the inclination angle of the storage restriction portion (upper surface) with respect to the horizontal plane was 0 °, 5 °, 10 °, 15 °, 20 °, 30 °, 35 °, and 45 °, respectively. . Other well plate specifications were as follows:
-Well width: 52.5 μm
-Well depth: 25 μm
-Number of wells: vertical 100 x horizontal 100 = 10,000-Containment restriction configuration: peninsula type-Number of containment restriction in each well: 6-Containment restriction width: 5 μm
-Length dimension of accommodation restriction part: 13.75 mm

  An aqueous dispersion containing beads was supplied to the manufactured well plate, and an attempt was made to accommodate the beads one by one in the well. Specifically, a dispersion obtained by dispersing zirconia beads having a diameter of 23 μm in water (the content of zirconia beads: 0.4 mg / 200 μl) was dropped with a dropper. When dropping, the dispersion was dropped so as to spread evenly in the 100 × 100 well formation region (no ultrasonic waves or long-term vibration was given). Through such an operation, the value of the accommodation rate at which the beads are accommodated in the wells was examined. In addition, operation was repeated 10 times and the average value was calculated | required from each accommodation rate.

The results are shown in Table 3 below, and a graph of the results is shown in FIG.
[Table 3]

  In Table 3, 0 ° is a state in which the accommodation restriction portion is not inclined and the upper surface of the accommodation restriction portion is horizontal. From the results of Table 3, it was found that the bead accommodation rate is clearly improved when the upper surface of the accommodation restriction portion is inclined so as to become lower toward the center. That is, it can be considered that the beads can be effectively guided to the center of the well when the beads settle, and the bead accommodation rate is improved. In addition, although the improvement effect of a housing rate appears notably in the range whose inclination angle with respect to a horizontal surface is 0 degree-30 degrees (especially 0 degree-20 degrees), the further improvement of a housing rate cannot be expected in the inclination angle beyond it. I understand.

(Example 2)
A test was conducted to confirm the effect of “a peninsula-shaped accommodation restriction portion having an inclined surface whose upper surface is inclined with respect to a horizontal plane”. First, a “well plate having an accommodation restriction portion inclined in the circumferential direction in each well” prepared by the above-described manufacturing method was prepared (see FIG. 13). Such a housing restriction portion whose height changes can be realized by using a halftone mask for exposure. Eight types of well plates were prepared so that the inclination angles of the inclined surface with respect to the horizontal plane were 0 °, 5 °, 10 °, 15 °, 20 °, 30 °, 35 °, and 45 °, respectively. Other well plate specifications were as follows:
-Well width: 52.5 μm
-Well depth: 25 μm
-Number of wells: vertical 100 x horizontal 100 = 10,000-Containment restriction configuration: peninsula type-Number of containment restriction in each well: 6-Containment restriction width: 5 μm
-Length dimension of accommodation restriction part: 13.75 mm
・ Inclination angle * of the restricted access part *: 20 °
*: The height of the accommodation restriction portion gradually decreases toward the center.

  An aqueous dispersion containing beads was supplied to the manufactured well plate, and an attempt was made to accommodate the beads one by one in the well. Specifically, a dispersion obtained by dispersing zirconia beads having a diameter of 23 μm in water (the content of zirconia beads: 0.4 mg / 200 μl) was dropped with a dropper. When dropping, the dispersion was dropped so as to spread evenly in the 100 × 100 well formation region (no ultrasonic waves or long-term vibration was given). Through such an operation, the value of the accommodation rate at which the beads are accommodated in the wells was examined. In addition, operation was repeated 10 times and the average value was calculated | required from each accommodation rate.

The results are shown in Table 4 below, and a graph of the results is shown in FIG.
[Table 4]

  In Table 4, 0 ° is a state in which the accommodation restriction portion is not inclined and the upper surface of the accommodation restriction portion is horizontal. From the results of Table 4, it was found that the bead accommodation rate was clearly improved when the upper surface of the accommodation limiting portion was inclined so as to be lowered along the circumferential direction. That is, it is considered that the beads can be effectively guided to the center of the well when the beads are settled, and the bead accommodation rate is improved. In particular, in consideration of the fact that the height of the accommodation restriction portion used in the present embodiment gradually decreases toward the center, the upper surface of the accommodation restriction portion is in the “radial direction” and “circumferential direction”. It was found that tilting from the horizontal plane in both points is desirable for improving the capacity.

(Example 3)
A test was conducted to confirm the effect of “a plurality of peninsula-shaped accommodation restriction portions whose height changes sequentially along the circumferential direction”. First, a “well plate having six accommodation restricting portions having different heights” prepared by the above-described manufacturing method was prepared (see FIG. 14). Such a housing restriction portion whose height changes can be realized by using a halftone mask for exposure. In addition, the height of the accommodation restricting portion in each well was 5 μm, 9 μm, 13 μm, 17 μm, 21 μm, and 25 μm on average based on the bottom surface inside the well. Other well plate specifications were as follows:
-Well width: 52.5 μm
-Well depth: 25 μm
-Number of wells: vertical 100 x horizontal 100 = 10,000-Containment restriction configuration: peninsula type-Number of containment restriction in each well: 6-Containment restriction width: 5 μm
-Length dimension of accommodation restriction part: 13.75 mm

  An aqueous dispersion containing beads was supplied to the manufactured well plate, and an attempt was made to accommodate the beads one by one in the well. Specifically, a dispersion obtained by dispersing zirconia beads having a diameter of 23 μm in water (the content of zirconia beads: 0.4 mg / 200 μl) was dropped with a dropper. When dropping, the dispersion was dropped so as to spread evenly in the 100 × 100 well formation region (no ultrasonic waves or long-term vibration was given). Through such an operation, the value of the accommodation rate at which the beads are accommodated in the wells was examined. In addition, operation was repeated 10 times and the average value was calculated | required from each accommodation rate. The same operation and the derivation of the accommodation rate were performed on the well plate in which the heights of the six accommodation restriction portions were all aligned.

The results are shown in Table 5 below.
[Table 5]

  From the results of Table 5, if the height of the storage restriction part is changed in each well (when “with a step”), the bead storage rate is higher than when all the heights are the same (when “without a step”). It turns out that it improves. This is presumably because the beads were able to move so as to slide down the “upper surface of the stepped accommodation restriction portion” when the beads settled by their own weight. That is, when the accommodation restriction part is formed in a stepped shape, the beads that settle can be effectively guided to the center part of the well, and the bead accommodation rate can be improved.

  The confirmation test of the accommodation effect was carried out not on the peninsula-like accommodation restriction part but on the pillar-like accommodation restriction part (Examples 4 to 6).

Example 4
A test was conducted to confirm the effect of the “pillar-shaped accommodation restriction portion whose height gradually decreases toward the center”. First, a “well plate having an accommodation restriction portion inclined in the central direction in each well” prepared by the above-described manufacturing method was prepared (see FIG. 19). Such a housing restriction portion whose height changes can be realized by using a halftone mask for exposure. In addition, eight types of well plates were prepared so that the inclination angles of the housing restriction portions with respect to the horizontal plane were 0 °, 5 °, 10 °, 15 °, 20 °, 30 °, 35 °, and 45 °, respectively. Other well plate specifications were as follows:
・ Width of well: 90μm
-Well depth: 25 μm
-Number of wells: 100 vertical x 100 horizontal = 10000-Form of storage restriction: pillar type-Number of storage restriction in each well: 4-Outer diameter of storage restriction: 50 μm
-Inner diameter dimension of the accommodation restriction part: 30 μm
・ Width of notch: 10μm

  For the manufactured well plate, a dispersion liquid in which beads having a diameter of 25 μm were dispersed in water was provided, and an attempt was made to accommodate the beads one by one in the well. Specifically, a dispersion obtained by dispersing zirconia beads having a diameter of 25 μm in water (the content of zirconia beads: 0.4 mg / 200 μl) was dropped with a dropper. When dropping, the dispersion was dropped so as to spread evenly in the 100 × 100 well formation region (no ultrasonic waves or vibration for a long time). As a result, the value of the accommodation rate at which the beads were accommodated in the wells was examined. The operation was repeated 10 times, and the average value was obtained from each accommodation rate.

The results are shown in Table 6 below, and a graph of the results is shown in FIG.
[Table 6]

  In Table 6, 0 ° is a state in which the accommodation restriction portion is not inclined and the upper surface of the accommodation restriction portion is horizontal. From the results of Table 6, it was found that the bead accommodation rate was clearly improved when the upper surface of the accommodation restriction portion was inclined so as to become lower toward the center. That is, it can be considered that the beads can be effectively guided to the central portion of the well when the beads are settled, and the bead accommodation rate is improved. In addition, it was also found that the improvement effect of the accommodation rate appears remarkably when the inclination angle with respect to the horizontal plane is in the range of 0 ° to 30 ° (that is, further improvement of the accommodation rate cannot be expected at an inclination angle larger than 30 °. I understood).

(Example 5)
A test was conducted to confirm the effect of the “pillar-shaped accommodation limiting portion having an inclined surface whose upper surface is inclined with respect to a horizontal plane”. Therefore, a “well plate provided with each circumferentially inclined accommodation restriction portion” prepared by the above-described manufacturing method was prepared (see FIG. 20). Such a housing restriction portion whose height changes can be realized by using a halftone mask for exposure. In addition, eight types of well plates were prepared so that the inclination angles of the housing restriction portions with respect to the horizontal plane were 0 °, 5 °, 10 °, 15 °, 20 °, 30 °, 35 °, and 45 °, respectively. Other well plate specifications were as follows:
・ Width of well: 90μm
-Well depth: 25 μm
-Number of wells: 100 vertical x 100 horizontal = 10000-Form of storage restriction: pillar type-Number of storage restriction in each well: 4-Outer diameter of storage restriction: 50 μm
-Inner diameter dimension of the accommodation restriction part: 30 μm
・ Width of notch: 10μm
・ Inclination angle ^{* of the} restricted access part: 20 °
*: The height of the accommodation restriction portion gradually decreases toward the center.

  An aqueous dispersion containing beads was supplied to the manufactured well plate, and an attempt was made to accommodate the beads one by one in the well. Specifically, a dispersion obtained by dispersing zirconia beads having a diameter of 25 μm in water (the content of zirconia beads: 0.4 mg / 200 μl) was dropped with a dropper. When dropping, the dispersion was dropped so as to spread evenly in the 100 × 100 well formation region (no ultrasonic waves or long-term vibration was given). Through such an operation, the value of the accommodation rate at which the beads are accommodated in the wells was examined. In addition, operation was repeated 10 times and the average value was calculated | required from each accommodation rate.

The results are shown in Table 7 below, and a graph of the results is shown in FIG.
[Table 7]

  In Table 7, 0 ° is a state in which the accommodation restriction portion is not inclined and the upper surface of the accommodation restriction portion is horizontal. From the results in Table 7, it was found that the bead accommodation rate was clearly improved when the upper surface of the accommodation restriction portion was inclined so as to be lowered along the circumferential direction. That is, it is considered that the beads can be effectively guided to the center of the well when the beads are settled, and the bead accommodation rate is improved. In particular, in consideration of the fact that the housing restriction portion used in the present embodiment gradually decreases in height toward the central direction, the upper surface of the pillar has both “radial direction” and “circumferential direction”. It was found that tilting from the horizontal plane at this point is desirable for improving the accommodation rate.

(Example 6)
A test was conducted to confirm the effect of “a plurality of pillar-shaped accommodation restriction portions whose height changes sequentially along the circumferential direction”. First, a “well plate having four accommodation limiting portions having different heights” prepared by the above-described manufacturing method was prepared (see FIG. 21). Such a housing restriction portion whose height changes can be realized by using a halftone mask for exposure. In addition, the height of the accommodation restriction portion in each well was 7 μm, 13 μm, 19 μm, and 25 μm on average, based on the bottom surface inside the well. Other well plate specifications were as follows:
・ Width of well: 90μm
-Well depth: 25 μm
-Number of wells: 100 vertical x 100 horizontal = 10000-Form of storage restriction: pillar type-Number of storage restriction in each well: 4-Outer diameter of storage restriction: 50 μm
-Inner diameter dimension of the accommodation restriction part: 30 μm
・ Width of notch: 10μm

  An aqueous dispersion containing beads was supplied to the manufactured well plate, and an attempt was made to accommodate the beads one by one in the well. Specifically, a dispersion obtained by dispersing zirconia beads having a diameter of 25 μm in water (the content of zirconia beads: 0.4 mg / 100 μl) was dropped with a dropper. When dropping, the dispersion was dropped so as to spread evenly in the 100 × 100 well formation region (no ultrasonic waves or long-term vibration was given). Through such an operation, the value of the accommodation rate at which the beads are accommodated in the wells was examined. In addition, operation was repeated 10 times and the average value was calculated | required from each accommodation rate. The same operation and the derivation of the accommodation rate were performed on well plates in which the heights of the four accommodation restriction portions were all aligned.

The results are shown in Table 8 below.
[Table 8]

  From the results in Table 8, if the height of the storage restriction portion is changed in each well (in the case of “step difference”), the bead retention rate is higher than that in the case of the same height (in the case of “no step difference”). It turns out that it improves. This is presumably because the beads were able to move so as to slide down the “upper surface of the stepped accommodation restriction portion” when the beads settled by their own weight. That is, when the accommodation restriction part is formed in a stepped shape, the beads that settle can be effectively guided to the center part of the well, and the bead accommodation rate can be improved.

<< Various treatments using plate-like containers >>
The treatment shown below was performed using the plate-shaped container of the present invention.

(Analysis, extraction and purification of target substance)-peninsula-like accommodation restriction portion While using well plate P1 (well center space diameter: about 26 μm, well depth: about 25 μm) as shown in FIG. Using the fixed zirconia particles B1 (D = 23 μm), the capture characteristics and detection characteristics of the target substance were confirmed. Biotinylated HRP was used as the target substance. Avidin immobilized on the particles can specifically bind to biotinylated HRP.

The capture characteristics and detection characteristics of the target substance were confirmed by the following operation. First, about 4 mg (= about 1.1 × 10 ⁵ particles) of particle B1 was placed in a 1.5 ml tube, and 200 μl of 10 mM PBS buffer (pH 7.2) was added. An appropriate amount of the dispersion was taken with a pipette so as to suck in all the particles settling on the bottom, and dropped onto the well plate P1. By pipetting or tilting the plate, it was possible to put one particle B1 into almost all wells (see FIG. 43).

  After removing the buffer using a pipette, 100 μl of biotinylated HRP with a concentration of 20 ng / ml was added and subjected to vibration. Thereafter, the liquid that did not fully enter the well was removed with a pipette. Subsequently, the mixture was allowed to stand for 30 minutes with occasional vibrations so that a binding reaction between “avidin immobilized on particles” and “biotinylated HRP” was caused. Thereafter, the well and the particles contained therein were washed 4 times with 100 μl of 10 mM PBS buffer (pH 7.2). After removing the PBS buffer (pH 7.2) from the well, 100 μl of TMB (tetramethylbenzidine) was added as a color former and subjected to vibration. Next, the liquid that did not fully enter the well was quickly removed with a pipette. About 30 minutes later, the well region was enlarged and photographed with an Olympus microscope BX-50. Thereby, the color development of the particles contained in each well was observed. As a result, it was confirmed that each well was colored substantially uniformly.

  From the above, it was found that one particle that specifically binds to the target substance can be put in each well, and the target substance can be captured independently for each particle. As a result, it was found that when the well plate of the present invention is used, the target substance can be extracted and purified, and the target substance can be detected and analyzed using a coloring substance or the like.

(Confirmation of capture characteristics of 1 cell / 1 well)-peninsula-shaped accommodation restriction portion As a cell model, resin particles B2 (Spherotech Inc. SPHERO Biotin-Polystyrene Particles) having a diameter of 6.7 μm was used and a well plate was used in FIG. Using a well plate P2 (well center space diameter: about 10 μm, well depth: about 12 μm) as shown, a test to confirm “capture characteristics of 1 cell / 1 well” was performed. If one cell can be accommodated in each well, it becomes possible to separate the cells, and detection and screening can be performed by performing the same processing as in the above-mentioned “1 particle / 1 well” experiment. .

The capture characteristics of 1 cell / 1 well were confirmed by the following operation. First, about 2 μl (= about 1.1 × 10 ⁵ ) of a dispersion of 1 w / v% cell model particle B2 was charged in a 1.5 ml tube, and 100 μl of 10 mM PBS buffer (pH 7.2) was added. The whole amount of the dispersion was taken with a pipette while stirring the particles by pipetting and dropped onto the plate P2. From time to time, the plate was vibrated, and particles B2 could be put into almost all wells one by one.

  From the above, it has been found that one cell can be accommodated in each well (particularly, a recess provided with a peninsular accommodation restriction portion). That is, it was shown that cell separation, detection, screening and the like are possible using the well plate of the present invention.

(Analysis, extraction and purification of target substance)-island-like accommodation restriction portion Well plate P3 having an island-like accommodation restriction portion as shown in Fig. 5 (well center space diameter: about 26 µm, well depth: about 25 µm) ) And zirconia particles B3 having avidin immobilized on the surface thereof were used to perform a test to confirm the capture characteristics and detection characteristics of the target substance. Biotinylated HRP was used as the target substance. Avidin immobilized on the particles can specifically bind to biotinylated HRP.

The capture characteristics and detection characteristics of the target substance were confirmed by the following operation. First, about 4 mg (= about 1.1 × 10 ⁵ particles) of particle B3 was placed in a 1.5 ml tube, and 200 μl of 10 mM PBS buffer (pH 7.2) was added. An appropriate amount of the dispersion was taken with a pipette so as to suck in all the particles settling on the bottom, and dropped onto the plate P3. By pipetting or tilting the plate, one particle B3 could be put in almost all wells.

  After removing the buffer using a pipette, 100 μl of biotinylated HRP with a concentration of 20 ng / ml was added and subjected to vibration. Thereafter, the liquid that did not fully enter the well was removed with a pipette. Subsequently, the mixture was allowed to stand for 30 minutes so as to cause a binding reaction between “avidin immobilized on the particles” and “biotinylated HRP” with occasional vibration. Thereafter, the well and the particles contained therein were washed 4 times with 100 μl of 10 mM PBS buffer (pH 7.2). After removing the PBS buffer (pH 7.2), 100 μl of TMB (tetramethylbenzidine) was added as a color former, and after shaking, the liquid that could not fit into the well was quickly removed with a pipette. About 30 minutes later, the well region was enlarged and photographed with an Olympus microscope BX-50. Thereby, the color development of the particles contained in each well was observed. As a result, it was confirmed that each well was colored substantially uniformly.

  From the above, it is possible to put one particle that specifically binds to the target substance into each well (especially, a well having an island-shaped accommodation restriction part), and each target substance is independent for each particle. It was found that can be captured. As a result, it was found that when the well plate of the present invention is used, the target substance can be extracted and purified, and the target substance can be detected and analyzed using a coloring substance or the like.

(Confirmation of capture characteristics of 1 cell / 1 well)-Island-like accommodation restriction portion As a cell model, resin particles B4 (Spherotech Inc. SPHERO Biotin-Polystyrene Particles) having a diameter of 6.7 μm were used and a well plate in FIG. Using a well plate P4 (well center space diameter: about 26 μm, well depth: about 25 μm) having island-shaped accommodation restriction portions as shown, the capture characteristics of 1 cell / 1 well were confirmed. First, about 2 μl (= about 1.1 × 10 ⁵ ) of a dispersion of 1 w / v% cell model particle B4 was charged in a 1.5 ml tube, and 100 μl of 10 mM PBS buffer (pH 7.2) was added. The whole amount of the dispersion was taken with a pipette while stirring the particles by pipetting and dropped onto the plate P4. From time to time, the plate was allowed to vibrate, so that one particle B4 could be put in almost every well.

  From the above, it has been found that one cell can be accommodated in each well (particularly, a recess having an island-shaped accommodation restriction portion). That is, it was shown that cell separation, detection, screening and the like are possible using the well plate of the present invention.

《Metallic coating effect confirmation test》
A test was conducted to confirm the effect of the metal coating that can be applied to the plate-like member. Specifically, an Au thin film was formed by sputtering, and a test for confirming the effect was performed.

First, an Au thin film was formed on the surface of the resin substrate by using a MAC240 sputtering apparatus manufactured by BAL-TEC. Specifically, three Au targets having a diameter of 54 mm and a thickness of 0.3 mm were used as targets. During film formation, the pressure in the chamber was reduced to 7 × 10 ⁻⁵ mbar, and then argon was introduced for 10 seconds. Sputtering was performed for 90 seconds at a sputtering current of 100 mA with an argon flow valve opening rate of 50%, and a 100 nm thin film was produced. For the measurement of the film thickness, P-12 Disk PRFILER manufactured by KLA Tencor was used.

By using “zirconia particles B5 having avidin immobilized on the surface” for the “plate P5 having an Au coating” thus obtained, the capture characteristics and detection characteristics of the target substance were confirmed. Biotinylated HRP was used as the target substance. First, about 4 mg (= about 1.1 × 10 ⁵ particles) of particle B5 was placed in a 1.5 ml tube, and 200 μl of 10 mM PBS buffer (pH 7.2) was added. An appropriate amount of the dispersion was taken with a pipette so as to suck in all the particles settling on the bottom, and dropped onto the plate P5. By pipetting or tilting the plate, one particle B5 could be put in almost all wells. After removing the buffer solution using a pipette, 100 μl of biotinylated HRP at a concentration of 20 ng / ml was added, and vibration was applied. Thereafter, the liquid that did not fully enter the well was removed with a pipette. Subsequently, the mixture was allowed to stand for 30 minutes so as to cause a binding reaction between “avidin immobilized on the particles” and “biotinylated HRP” with occasional vibration. The wells and the contained particles were then washed 4 times with 100 μl of 10 mM PBS buffer (pH 7.2). After removing the PBS buffer (pH 7.2), 100 μl of TMB (tetramethylbenzidine) was added as a color former, and after shaking, the liquid that could not fit into the well was quickly removed with a pipette. About 30 minutes later, the well region was enlarged and photographed using an Olympus microscope BX-50. Thereby, the color development of the particles contained in each well was observed. As a result, it was confirmed that each well was colored substantially uniformly. In particular, since the Au thin film was produced on the plate P5, there was no color transfer from the adjacent well, and the color development in each well could be clearly observed. That is, the color development in each well could be observed separately.

  When the plate-like container of the present invention is used, target substance can be analyzed, extracted or purified, and cells can be separated, detected or screened (particularly, such treatment / operation can be carried out in large quantities at once). Therefore, the plate-like container of the present invention can be used for gene analysis, expression analysis, protein analysis, antigen / antibody reaction analysis, cell analysis and the like.

  More specifically, taking into account the effects of the present invention, the plurality of micro reaction vessel portions that are the plate-like containers of the present invention are capable of freely designing the volume of the solution used for (i) reaction or culture. And (ii) a structure in which only one granular material is accommodated, and (iii) the granular material jumps out to the outside once the granular material enters the inside of the minute reaction vessel part by natural sedimentation or the like. It has the effect of “hard”. Therefore, the particulate matter is sufficiently immersed in the solution charged in the minute reaction container, which is suitable for separation, purification, extraction, analysis, inspection and the like performed in the fields of medical science and bioscience.

FIG. 1 is a perspective view schematically showing an embodiment of a plate-like container and a recess according to the present invention. FIG. 2 is a top view (FIG. 2A) and a perspective view (FIG. 2B) schematically showing an aspect of a peninsula-like accommodation limiting portion. FIGS. 3A and 3B are a top view (FIG. 3A) and a perspective view (FIG. 3B) schematically showing an aspect of the pillar-shaped accommodation limiting portion. FIG. 4 schematically shows a cross-sectional view of the pillar-shaped accommodation limiting portion shown in FIG. FIG. 5 is a top view (FIG. 5 (a)) and a perspective view (FIG. 5 (b)) schematically showing an aspect of the island-shaped accommodation restriction portion. FIG. 6 is a top view (FIG. 6 (a)) and a perspective view (FIG. 6 (b)) schematically showing an embodiment provided with two peninsula-shaped accommodation limiting portions extending out of the radial direction. is there. FIGS. 7A and 7B are a top view (FIG. 7A) and a perspective view (FIG. 7B) schematically showing a mode in which two peninsula-like accommodation limiting portions extending along the radial direction are provided. is there. FIG. 8 is a top view (FIG. 8 (a)) and a perspective view (FIG. 8 (b)) schematically showing an embodiment provided with five peninsula-shaped accommodation restriction portions. FIG. 9 is a top view (FIG. 9 (a)) and a perspective view (FIG. 9 (b)) schematically showing an embodiment provided with six peninsula-shaped accommodation restriction portions. FIG. 10 is a top view (FIG. 10 (a)) and a perspective view (FIG. 10) schematically showing an aspect formed by replacing the “accommodation restricting portion” and the “liquid containing portion” in FIG. (B)). FIG. 11 is a top view (FIG. 11 (a)) and a perspective view (FIG. 11 (b)) schematically showing an aspect in which a “liquid reservoir” is provided adjacent to the accommodation restricting portion. FIG. 12 is a perspective view schematically showing an embodiment of a peninsula-shaped accommodation limiting portion that inclines toward the center of the recess. FIG. 13 is a perspective view schematically showing an aspect of a peninsula-shaped accommodation restricting portion inclined along the circumferential direction. FIG. 14 is a perspective view schematically showing an aspect of a plurality of peninsula-shaped accommodation restricting portions whose heights are different in stages. FIG. 15 is a top view (FIG. 15 (a)) and a perspective view (FIG. 15 (b)) schematically showing an aspect in which the accommodation limiting portion is provided so as to extend spirally from the peripheral portion of the recess. It is. FIGS. 16A and 16B are a top view (FIG. 16A) and a perspective view (FIG. 16B) schematically showing an aspect including two pillar-shaped accommodation limiting portions. FIGS. 17A and 17B are a top view (FIG. 17A) and a perspective view (FIG. 17B) schematically illustrating an aspect including three pillar-shaped accommodation limiting portions. FIG. 18 is a top view (FIG. 18 (a)) and a perspective view (FIG. 18 (b)) schematically showing an aspect provided with four pillar-shaped accommodation limiting portions. FIG. 19 is a perspective view schematically showing an aspect of a pillar-shaped accommodation limiting portion that inclines toward the center of the recess. FIG. 20 is a perspective view schematically illustrating an aspect of a pillar-shaped accommodation limiting portion that is inclined along the circumferential direction. FIG. 21 is a perspective view schematically showing an aspect of a plurality of pillar-shaped accommodation restricting portions whose heights are different in stages. FIG. 22 is a top view (FIG. 22 (a)) and a perspective view (FIG. 22 (b)) schematically showing an aspect in which the pillar-shaped accommodation limiting portion is provided so as to extend spirally. . FIG. 23 is a top view (FIG. 23 (a)) and a perspective view (FIG. 23 (b)) schematically showing an embodiment in which four cylindrical “pillars” are provided in each recess. FIG. 24 is a top view (FIG. 24 (a)) and a perspective view (FIG. 24 (b)) schematically showing an aspect provided with three “island-like” accommodation restriction portions. FIG. 25 is a top view (FIG. 25 (a)) and a perspective view (FIG. 25 (b)) schematically showing an aspect provided with four “island-like” accommodation restriction portions. FIG. 26 is a perspective view schematically showing a modified example of the “island-like” accommodation restriction portion. FIG. 27 is a perspective view schematically showing a modified example of the “island-like” accommodation restriction portion. FIG. 28 is a perspective view schematically showing a modified example of the “island-like” accommodation restriction portion. FIG. 29 is a top view (FIG. 29 (a)) and a perspective view (FIG. 29 (b)) schematically showing a cylindrical well as found in the prior art. FIG. 30 shows a state in which two spherical particles are housed in a cylindrical well, and is a schematic diagram showing a state in which half of one particle is housed in the well. FIG. 31 is a top view (FIG. 31 (a)) and a perspective view (FIG. 31 (b)) schematically showing a quadrangular prism-shaped well as found in the prior art. FIG. 32 shows a state where two spherical particles are accommodated in a quadrangular prism-shaped well, and is a schematic diagram showing a state where half of one particle is accommodated in the well. FIG. 33A is a diagram schematically showing an example of a mask pattern arranged on a resist, and FIG. 33B is an overall shape of a plate-like container in which a “well formation region” is represented. FIG. FIG. 34 is a diagram schematically showing a modified embodiment of the plate-shaped container of the present invention. FIG. 35 is a diagram schematically showing a modified embodiment of the plate-shaped container of the present invention. FIG. 36 is a diagram schematically showing a modified embodiment of the plate-shaped container of the present invention. FIG. 37 is an electron micrograph of the well formation region of the plate-like container prepared in the example of the present invention. FIG. 38 is a graph showing the results of Example 1. FIG. 39 is a graph showing the results of Example 2. FIG. 40 is a graph showing the results of Example 4. FIG. 41 is a graph showing the results of Example 5. FIG. 42 is a perspective view schematically showing an aspect of the plate-like container used in the examples. FIG. 43 is an optical micrograph showing the result of storing one particle in each well.

Explanation of symbols

25 (25a to 25d) accommodation limiting portion 30 recess (or well)
30 'cylindrical well 30''square column shaped well 30a containing object A containing area 30b containing object B containing area 30b' liquid reservoir 50 plate-like member 60 spherical particle 70 mask 100 plate-like container D bead or Diameter of spherical particles such as cells P Notch in the pillar-shaped accommodation restriction

Claims (16)

A plate-like container in which a plurality of wells are formed in a plate-like member so that both a solid-like object A and a liquid-like object B are accommodated,
While accommodating the contained object B to each of the wells, accommodating limiting portion for accommodating the contained object A, one for each of said well are formed in the interior of the well,
Each of the wells has a width dimension and a depth dimension that are larger than the diameter dimension D of the object A, and the accommodation limiting portion is provided so as to occupy a part of the internal space of each well. ,Also
The internal space of the well excluding the accommodation restricting portion and the accommodation space for the object to be accommodated A constitutes an accommodation space (volume Vb) for the object to be accommodated B. To be accommodated when the object to be accommodated B and the “spherical object to be accommodated A having the diameter D” are accommodated in the cylindrical space having the same bottom diameter and height as those in FIG. When the volume of B is the minimum accommodation volume Vb ′, the volume Vb is 9 to 30 times the minimum accommodation volume Vb ′ . 2. The plate-like container according to claim 1, wherein the accommodation limiting portion is integrated with at least a part of a peripheral portion of the well and extends from the peripheral portion.   The plate-shaped container according to claim 2, wherein a horizontal cross section of the accommodation limiting portion is rectangular or trapezoidal. The plate-like container according to claim 1, wherein the accommodation restricting portion has a form separated from a peripheral portion of the well .   The plate-shaped container according to claim 4, wherein the accommodation limiting portion is provided so as to at least partially surround the accommodation space of the object to be accommodated A. Gap dimension is formed between the peripheral portion of the said housing limiting unit well, as the peripheral portion of the well along the helical direction towards the center, characterized in that gradually widened, The plate-shaped container according to claim 4. The plate-like container according to any one of claims 1 to 6, wherein 2 to 6 of the accommodation restriction portions are provided for each of the wells . The plate-shaped container according to any one of claims 1 to 7, wherein a height of the accommodation restriction portion is gradually lowered from a peripheral portion of the well toward the center.   The plate-shaped container according to any one of claims 1 to 8, wherein an upper surface of the accommodation restricting portion forms an inclined surface inclined with respect to a horizontal plane. Each of the wells is provided with a plurality of storage restriction portions,
10. Each of the wells has a different height from each other so that the height of the accommodation restriction portion sequentially changes along a circumferential direction. The plate-shaped container in any one of. Wherein the pre-Symbol plate-shaped container is well plate or microarray, the plate-shaped container according to any one of claims 1 to 10.   The plate-like container according to claim 11, wherein the plate-like member is provided with a metal coating.   The plate-shaped container according to claim 11, wherein the plate-shaped member has hydrophilicity.   A metal mold used for forming the plate-like container according to claim 1.   Each of the wells provided in the plate-like container according to any one of claims 11 to 13 contains "beads to which a target substance can bind", and the target substance is analyzed in each of the wells. A method of extraction or purification.   A method for separating, detecting or screening the cells by storing cells one by one in the well provided in the plate-like container according to claim 11.