Microplates

Abstract

To provide a microplate suitable for an automatic culture apparatus, which makes it easy to focus on cultured cells in each well in automatic observation and has excellent observation efficiency.SOLUTION: A microplate 1 comprises: an upper surface portion 10 having a rectangular plan view shape; a peripheral wall portion 12 vertically hung from the outer edge portion of the upper surface portion 10; and a plurality of bottomed cylindrical wells 14 in which a circular opening is formed in an upper surface 10a of the upper surface portion 10. Regarding the deepest parts of two wells 14, which are farthest from each other, the distance d1 between the deepest parts of two wells 14 is 400 μm or less in the direction perpendicular to a reference plane calculated by a reference plane calculation method.SELECTED DRAWING: Figure 2

Description

[Technical field]

[0001] The present invention relates to a microplate. [Background technology]

[0002] Cultured cells are often used as specimens in basic research to elucidate life phenomena and drug discovery research, and culture vessels are widely used to obtain large amounts of specimens. Conventional evaluation of cultured cells has mainly been based on characteristics of cell metabolism or metabolites, such as MTT assay and ELISA (Enzyme-Linked Immuno Sorbent Assay). On the other hand, three-dimensional cell masses (spheroids) obtained by 3D culture, in which cells are aggregated, have a three-dimensional structure similar to that in vivo. For this reason, the importance of morphological observation using cell masses has increased in recent years.

[0003] As a culture vessel used for three-dimensional culture, a microplate is known that has multiple wells, each of which has a bottom surface (culture surface) on which many fine wells (microwells) with a pore size of 100 to 1,000 μm are formed (Patent Document 1). When cells are seeded on the culture surface, the cells meet in each of the fine wells to form cell clusters. The ANSI (American National Standards Institute) / SBS standard is known as a standard for microplates. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Patent No. 6400575 Summary of the Invention [Problem to be solved by the invention]

[0005] As a cell culture method, a method for culturing a large amount of cells at high throughput using an automated culture device has been developed. However, with conventional microplates, even if they meet the ANSI / SBS standard, if the height of the container bottom varies, it is difficult to accurately adjust the amount of cells and culture solution collected from each well, which can reduce the uniformity of the evaluation system within the same plate.

[0006] In addition, for cell observation, a method has been developed to observe a large number of cells at high throughput using an automated observation device. However, with conventional microplates, even those that meet the ANSI / SBS standard, it is difficult to automatically focus on the cultured cells in each well, which can reduce the efficiency of automated observation.

[0007] An object of the present invention is to provide a microplate which is suitable for an automatic culture device and which allows easy focusing on cultured cells in each well during automatic observation, thereby achieving excellent observation efficiency. [Means for solving the problem]

[0008] The present invention has the following aspects. [1] A microplate having a plurality of wells, wherein the distance between the deepest points of the two wells that are furthest apart in a direction perpendicular to a reference plane calculated by the reference plane calculation method described below is 400 μm or less. (Reference surface calculation method) Of all the wells, four wells are selected that are located at the vertices of a square or rectangle in plan view and have the largest area of ​​the square or rectangle. Of the four wells, an xy plane is set that passes through the deepest parts of two wells that are diagonally located in plan view (however, if the bottom of the well is flat, the center of the bottom of the well in plan view), the midpoint of the deepest parts of the two wells is the origin, and the xy coordinates of the deepest parts of the four wells are (-X, Y), (X, Y), (-X, -Y), and (X, -Y), respectively. In the three-dimensional orthogonal coordinate system (x, y, z), when the coordinates of the deepest parts of the four wells are (-X, Y, Z1), (X, Y, Z2), (-X, -Y, Z3), and (X, -Y, Z4), the planes represented by the following formulas 1 to 4 are used as the reference plane.

[0009]

number

[0010] [2] A microplate having a plurality of wells, wherein the distance between the reference plane calculated by the reference plane calculation method and the bottom of each well is 200 μm or less. [3] A microplate according to [1] or [2], wherein the plurality of wells are arranged in a rectangular matrix of n x m pieces in a planar view, and the reference plane is calculated by the reference plane calculation method using four wells located at the four corners of the plurality of wells. [4] The microplate described in any of [1] to [3], wherein the cross-sectional shape of the bottom surface of the well when cut along a plane parallel to the depth direction passing through the center of the bottom surface in a planar view is a downwardly convex arc, a downwardly convex broken line, or a straight line. [5] A microplate according to any of [1] to [4], comprising a frame and a flat base material, the frame having a plurality of tubular portions forming peripheral walls of each of the wells, and the flat base material being attached to the lower ends of the plurality of tubular portions to form the plurality of wells. [6] The microplate according to any one of [1] to [5], wherein a plurality of microwells are formed in the bottom surface of the well. Effect of the Invention

[0011] According to the present invention, it is possible to provide a microplate which is suitable for an automatic culture device, and which is excellent in observation efficiency because it is easy to focus on cultured cells in each well during automatic observation. [Brief description of the drawings]

[0012] [Figure 1] FIG. 2 is a plan view of a microplate according to an embodiment. [Diagram 2] FIG. 2 is a cross-sectional view of the microplate of FIG. 1. [Diagram 3] FIG. 11 is an explanatory diagram for explaining a reference plane calculation method. [Figure 4] FIG. 2 is an enlarged cross-sectional view showing the bottom surface of each well of a microplate. [Diagram 5] FIG. 11 is a cross-sectional view of a microplate according to another embodiment. [Figure 6] FIG. 11 is a cross-sectional view of a microplate according to another embodiment. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] The meanings and definitions of terms used in this specification are as follows. A numerical range expressed by "to" means that the numerical values ​​before and after the "to" are the lower and upper limits of the numerical range. The "deepest part of the well" means the deepest point on the bottom surface (culture surface) of the well, and when the bottom surface of the well is flat, it refers to the center of the bottom surface of the well when viewed from above.

[0014] An example of an embodiment of the microplate of the present invention will be described below with reference to the drawings. Note that the dimensions of the drawings shown in the following description are merely examples, and the present invention is not necessarily limited thereto, and can be appropriately modified and implemented within the scope of the present invention.

[0015] [First embodiment] As shown in Figures 1 and 2, the microplate 1 of this embodiment has an upper surface portion 10 that is rectangular in plan view, a peripheral wall portion 12 that hangs down vertically from the outer edge of the upper surface portion 10, and a plurality of bottomed cylindrical wells 14 that have circular openings formed on the upper surface 10a of the upper surface portion 10.

[0016] In the microplate 1, the bottom surface 16 of each well 14 serves as a culture surface. The cross-sectional shape of the bottom surface 16 of the well 14 in this example when cut along a plane parallel to the depth direction passing through the center of the bottom surface 16 in a plan view is linear. That is, the well 14 in this example has a flat bottom with a flat bottom surface 16. The cross-sectional shape of the bottom surface 16 of the well 14 when cut along a plane parallel to the depth direction passing through the center of the bottom surface 16 in a plan view is not limited to a linear shape, and may be a downwardly convex arc shape, a downwardly convex broken line shape, or the like.

[0017] The arrangement pattern of the wells 14 in the microplate 1 is not particularly limited, and examples thereof include a pattern in which the wells are arranged vertically and horizontally in a matrix of n×m rectangles, and a staggered pattern. The opening shape of the wells 14 in a plan view is not limited to a circle, and may be, for example, a rectangle. The number of wells 14 is not particularly limited, and examples thereof include 6 to 1536 wells. In the microplate 1 of this example, 96 wells 14 are arranged in a rectangular matrix of 8 x 12 in plan view. However, it may alternatively be 16 x 24 = 384 wells, or 32 x 48 = 1536 wells.

[0018] In microplate 1, the distance d1 between the deepest points of two wells 14 that are furthest apart in a direction perpendicular to a reference plane calculated by a reference plane calculation method described below is 400 μm or less.

[0019] (Reference surface calculation method) Of all the wells 14, four wells 14 are selected that are located at the vertices of a square or rectangle in a plan view and that maximize the area of ​​the square or rectangle. In this example, as shown in FIG. 1, of the 96 wells 14 arranged in a rectangular matrix of 8×12 vertically and horizontally, the rectangle P with the largest area is the rectangle in which the four wells 14a to 14d located at the four corners are located at the vertices.

[0020] Then, in the microplate 1, a three-dimensional orthogonal coordinate system (x, y, z) is set that satisfies the following three conditions (1) to (3). (1) Of the four wells 14a to 14d, the wells have an xy plane that passes through the deepest parts of the two wells 14b and 14c that are diagonally positioned in a plan view. (2) The midpoint of the deepest parts of the two diagonally opposite wells 14b, 14c is the origin. (3) The x and y coordinates of the deepest parts of the four wells 14a to 14d are (-X, Y), (X, Y), (-X, -Y), and (X, -Y), respectively. However, when the bottom surface of the well 14 is flat as in this example, the deepest part of the well 14 is the center of the bottom surface 16 of the well 14 in a plan view.

[0021] In this three-dimensional orthogonal coordinate system (x, y, z), the coordinates of the deepest parts of the four wells 14a to 14d are A (-X, Y, Z1), B (X, Y, Z2), C (-X, -Y, Z3), and D (X, -Y, Z4), respectively. A plane that passes through the deepest parts (points B and C) of the diagonally positioned wells 14b and 14c among the four wells 14a to 14d and has the smallest distance to the deepest parts (points A and D) of the remaining two wells 14a and 14d is set as a reference plane.

[0022] Specifically, in the three-dimensional orthogonal coordinate system (x, y, z) shown in FIG. 3, an equation expressing a reference plane k is given by the following formula 1. z=ax+by+c Equation 1

[0023] When the plane expressed by equation 1, which passes through points B and C, is moved around balance axis m, which passes through points B and C, the plane with the smallest distance to points A and D is set as the reference plane k. Since the surface expressed by equation 1 passes through points B and C, substituting the coordinates of points B and C into equation 1 yields the following equations 5 and 6, respectively. Z2=aX+bY+c Equation 5 Z3=-aX-bY+c Equation 6

[0024] Adding together equations 5 and 6, we get equation 4 below. Z2+Z3=2c c=(Z2+Z3) / 2...Equation 4

[0025] Taking the difference between equation 5 and equation 6, we obtain equation 3 below. Z2-Z3=2aX+2bY 2bY=(Z2-Z3)-2aX b={(Z2-Z3) / 2-aX} / 2Y b=1 / Y{(Z2-Z3) / 2-aX}...Equation 3

[0026] Substituting equations 3 and 4 into equation 1, we obtain equation 7 below. Z=ax+1 / Y{(Z2-Z3) / 2-aX}y+(Z2+Z3) / 2...Equation 7

[0027] On the reference plane k, let point A'(-X, Y, Z1') be the point whose x and y coordinates are the same as those of point A. Substituting the coordinates of point A' into equation 7, we obtain equation 8 below. Z1'=-aX+1 / Y{(Z2-Z3) / 2-aX}Y+(Z2+Z3) / 2 Z1'=-2aX+Z2...Formula 8

[0028] Similarly, on the reference plane k, a point whose x and y coordinates are the same as those of point D is defined as point D'(X, -Y, Z3'). Substituting the coordinates of point D' into equation 7, the following equation 9 is derived. Z4'=aX-1 / Y{(Z2-Z3) / 2-aX}Y+(Z2+Z3) / 2 Z4'=2aX+Z3...Equation 9

[0029] When the distance between the surface represented by equation 1 (the surface represented by equation 7) and points A and D is minimum, the sum of the distance between points A and A' and the distance between points D and D' in the z-axis direction is minimum. Taking the square of the difference in the z coordinates of points A and A' and substituting equation 8 gives us the following equation 10. Similarly, taking the square of the difference in the z coordinates of points D and D' and substituting equation 9 gives us the following equation 11. (Z1-Z1') 2 =(Z1+2aX-Z2) 2 ...Formula 10 (Z4-Z4') 2 =(Z4-2aX-Z3) 2 ...Formula 11

[0030] Adding Equation 10 and Equation 11 together gives Equation 12. (Z1+2aX-Z2) 2 +(Z4-2aX-Z3) 2 =8a 2 X 2 +4aX(Z1-Z2-Z4+Z3)+(Z1-Z2) 2 +(Z4-Z3) 2 =8X 2 {a 2 +(1 / 2X)×(Z1-Z2-Z4+Z3)a}+(Z1-Z2) 2 +(Z4-Z3) 2 ...Formula 12

[0031] If Q = (1 / 2X) × (Z1-Z2-Z4+Z3), equation 12 can be expressed as equation 13 below. 8X 2 {a 2 +(a / 2X)×(Z1-Z2-Z4+Z3)}+(Z1-Z2) 2 +(Z4-Z3) 2 =8X 2 {a 2 +aQ}+(Z1-Z2) 2 +(Z4-Z3) 2 =8X 2{(a+Q / 2) 2 -Q 2 / 4}+(Z1-Z2) 2 +(Z4-Z3) 2 ...Formula 13

[0032] Formula 13 is smallest, that is, the distance between the surface expressed by formula 1 and points A and D is smallest when a+Q / 2=0, so a=-Q / 2. Substituting Q=(1 / 2X)×(Z1-Z2-Z4+Z3) into this, we derive formula 2 below. a=-1 / 2×(1 / 2X)×(Z1-Z2-Z4+Z3) Equation 2

[0033] From the above, the reference plane k is a plane expressed by formulas 1-4.

[0034]

number

[0035] In the microplate 1, the distance d1 between the deepest parts of the two wells 14 that are furthest apart in the direction perpendicular to the reference plane k is 400 μm or less, preferably 200 μm or less, and more preferably 50 μm or less. If the distance d1 is equal to or less than the upper limit, the deviation in height of the bottom surface 16 (culture surface) of each well 14 is small, so that when applied to an automatic culture device, it is easy to match the amount of cells and culture solution recovered from each well, and the uniformity of the evaluation system within the same plate is improved. In addition, the deviation in height of the cultured cells cultured in each well 14 is small, making it easy to focus on each cultured cell even in high-throughput observation using an automatic observation device, and improving observation efficiency.

[0036] As shown in FIG. 4, a plurality of uniformly sized fine wells 19 are preferably formed on the bottom surface 16 of the well 14, as this makes it easier to obtain cell clusters of uniform size. The shape of the opening of the micro wells 19 is not particularly limited, and may be, for example, a circle, an ellipse, a polygon, or an irregular shape. The dimensions of the opening of the micro wells 19, such as the average diameter, opening area, and depth, are not particularly limited and may be set appropriately. For example, the average diameter of the opening of the micro wells 19 is 100 to 2500 μm, and the opening area is 7850 to 4906250 μm. 2 The number of fine wells 19 formed on the bottom surface 16 per unit area is set to 10 to 10,000 pieces / cm. 2 can be set to.

[0037] The arrangement pattern of the microwells 19 is not particularly limited, and may be formed in a regular pattern, may be formed irregularly, or may have a mixture of regular and irregular parts. Examples of regular arrangement patterns include a pattern in which microwells are arranged at each vertex of a square arranged without gaps, a pattern in which microwells are arranged at each vertex and the center of a regular hexagon arranged without gaps, and a staggered pattern.

[0038] The material of the microplate 1 can be, for example, a resin. Examples of the resin include polystyrene resin, polyester resin, polyethylene resin, polypropylene resin, acrylic resin, polycarbonate resin, and silicone resin. The resin is preferably one selected from polystyrene resin, polyester resin, polyethylene resin, polypropylene resin, acrylic resin, polycarbonate resin, and silicone resin, and polystyrene resin is particularly preferable because of its high transparency and low drug adsorption. The resin constituting the microplate 1 may be one type or two or more types.

[0039] The peripheral wall 12 of the microplate 1 may be transparent or opaque, preferably opaque from the viewpoint of observational ease, and more preferably black in terms of color tone. The method for making the peripheral wall opaque is not particularly limited, and may be, for example, a method of adding fine particles or a method of adding a pigment.

[0040] The method for producing the microplate 1 is not particularly limited, and the microplate 1 can be molded by, for example, injection molding or compression molding. Among them, compression molding is preferred because it is easy to produce a microplate 1 having a plurality of wells 14 with a distance d1 of 400 μm or less.

[0041] An example of a method for forming the microwells 19 is laser irradiation. When the culture surface 18, which is the bottom surface 16 of the wells 14, is irradiated with laser light, the resin that constitutes the culture surface 18 melts and vaporizes, forming the microwells 19 with a very smooth surface. Around the openings of the microwells 19, the dissolved resin may rise up to form banks.

[0042] The laser light source is not particularly limited, and may be, for example, a CO2 laser. The arrangement and size of the fine wells 19 can be adjusted by adjusting the irradiation conditions such as the irradiation position and output of the laser light. For example, the size of the fine wells can be adjusted by setting the laser output to 1 to 100 W and the irradiation time to 0.1 to 100 μs.

[0043] A low adhesion coating film that suppresses cell adhesion may be formed on the bottom surface 16 of the well 14, which is the culture surface. Formation of the low adhesion coating film makes it easier to remove the cultured cells. The low adhesion coating film can be formed, for example, by applying a cell adhesion inhibitor. Examples of cell adhesion inhibitors include phospholipid polymers (2-methacryloyloxyethyl phosphorylcholine, etc.), polyhydroxyethyl methacrylate, fluorine-containing compounds, and polyethylene glycol. One type of cell adhesion inhibitor may be used alone, or two or more types may be used in combination. If the microplate 1 is made of a resin that has a cell adhesion suppressing effect, such as a silicone resin, it is possible to suppress adhesion of cells to the bottom surface 16 without forming a low adhesion coating film.

[0044] An easy-adhesion coating film that makes it easier for cells to adhere may be formed on the bottom surface 16 of the well 14. Examples of materials for forming the easy-adhesion coating film include collagen and gelatin. As the materials for forming the easy-adhesion coating film, one type may be used alone, or two or more types may be used in combination.

[0045] [Second embodiment] An example of the microplate of the second embodiment is the microplate 1 similar to that of the first embodiment. The microplate 1 of the second embodiment is similar to that of the first embodiment, except that the distance d2 between the reference plane k calculated by the above-mentioned reference plane calculation method and the bottom (deepest part) of each well 14 is 200 μm or less. "The distance d2 is 200 μm or less" means that all of the wells 14 in the microplate 1 satisfy the condition that the distance d2 between the bottom of the well 14 and the reference plane k is 200 μm or less.

[0046] The distance d2 between the reference plane k and the bottom (deepest part) of each well 14 is 200 μm or less, preferably 150 μm or less, and more preferably 50 μm or less. If the distance d2 is equal to or less than the upper limit, the deviation in height of the bottom surface 16 (culture surface) of each well 14 is small, so that when applied to an automatic culture device, it is easy to match the amount of cells and culture solution recovered from each well, and the uniformity of the evaluation system within the same plate is improved. In addition, the deviation in height of the cultured cells cultured in each well 14 is small, making it easy to focus on each cultured cell even in high-throughput observation using an automatic observation device, and improving observation efficiency.

[0047] A preferred aspect of the microplate 1 of the second embodiment is similar to that of the first embodiment, except for the distance d2. Examples of a method for manufacturing the microplate 1 of the second embodiment include the same method as that of the first embodiment, and compression molding is preferred because it is easy to manufacture a microplate 1 having a plurality of wells 14 with a distance d2 of 200 μm or less.

[0048] As described above, in the microplate of the present invention, in each well, the distance d1 is controlled to 400 μm or less, or the distance d2 is controlled to 200 μm or less. This reduces the deviation of the bottom surface 16 of each well 14, and when applied to an automatic culture device, it is easy to adjust the amount of cells and culture solution collected from each well, improving the uniformity of the evaluation system within the same plate. In addition, even with an automatic observation device, it becomes easy to automatically focus on the cultured cells cultured in each well 14. Therefore, a large number of cultured cells can be efficiently observed with high throughput by the automatic observation device. In the present invention, it is preferable that the distance d1 is controlled to 400 μm or less, and the distance d2 is controlled to 200 μm or less.

[0049] The microplate of the present invention is not limited to the above-mentioned embodiment. For example, the microplate of the present invention may be the microplate 2 illustrated in Fig. 5. The microplate 2 has the same configuration as the microplate 1 except that the wells 14 have a bottom surface 16A instead of the bottom surface 16. In Fig. 5, the same parts as those in Fig. 2 are denoted by the same reference numerals and the description thereof will be omitted.

[0050] The cross-sectional shape of bottom surface 16A of well 14 of microplate 2 when cut by a plane parallel to the depth direction passing through the center of bottom surface 16A in a plan view is a downwardly convex arc shape. In microplate 2, too, by controlling distance d1 to 400 μm or less, or distance d2 to 200 μm or less, or by satisfying both, it becomes easy for even an automatic observation device to automatically focus on the cultured cells cultured in each well 14.

[0051] The microplate of the present invention may be a microplate 3 exemplified in Fig. 6. In Fig. 6, the same parts as in Fig. 2 are given the same reference numerals and the explanation thereof will be omitted. The microplate 3 comprises a frame 20 and a flat base material 22. The frame 20 comprises an upper surface portion 10, a peripheral wall portion 12 vertically hanging down from the outer edge of the upper surface portion 10, and a circular opening formed on the upper surface 10a of the upper surface portion 10, and a plurality of tubular portions 24 each having an open lower end. The flat base material 22 is attached to the lower end sides of the plurality of tubular portions 24, thereby forming a plurality of wells 14.

[0052] Thus, microplate 3 has the same configuration as microplate 1, except that flat plate substrate 22, which is a separate member, is attached to the lower end side of each cylindrical portion 24 to form each well 14. In microplate 3, too, by controlling distance d1 to 400 μm or less, or controlling distance d2 to 200 μm or less, or by satisfying both, it becomes easy to adjust the amount of cells and culture solution to be collected from each well when applied to an automatic culture device, and it becomes easy to automatically focus on the cultured cells cultured in each well 14 even in an automatic observation device.

[0053] In addition, within the scope of the invention, the components in the above-described embodiments may be replaced with well-known components, and the above-described modified examples may be combined as appropriate. EXAMPLES

[0054] The present invention will be described in detail below with reference to examples, but the present invention is not limited to the following description. Example 1 is an example, and Example 2 is a comparative example.

[0055] [Example 1] A 96-well (8 × 12) microplate with a well bottom (culture surface) diameter of 6.45 mm was prepared in the manner illustrated in Figures 1 and 2, and the distances d1 and d2 were measured for each well. The distance d1 was 155 μm, and the maximum distance d2 was 89 μm.

[0056] [Example 2] A 6-well (8 × 12) microplate with a well bottom (culture surface) diameter of 6.45 mm was prepared in the manner illustrated in Figures 1 and 2, and the distance d1 and distance d2 were measured for each well. The distance d1 was 1350 μm, and the maximum value of distance d2 was 1320 μm.

[0057] [Accuracy of medium replacement in automated culture equipment] In a 96-well (8×12) microplate as shown in FIG. 1 and FIG. 2, 100 μL to 200 μL of medium is usually added for culturing. Usually, during culturing, the medium that supplies nutrients to the cells is replaced during the culturing process. In an automatic culturing device, the suction nozzle is set to a desired height (several μm above the culture surface) and the medium is replaced. When the suction nozzle is placed in accordance with the highest culture surface in the microplate and the medium in each well is aspirated, if there is a difference in the height of the deepest part of each well of the microplate, the amount of medium recovered will differ, causing differences in the cultures carried out under the same conditions and making it impossible to perform an accurate evaluation. Therefore, 200 μL of medium was added to each well of the microplate in each example, the suction nozzle of the automatic dispenser was set to match the deepest part of the well with the largest z value in the deepest part in the microplate, the medium was aspirated, and the remaining amount of medium in the well with the smallest z value in the deepest part in the microplate was calculated using the following formula. The volume error when 200 μL of new medium was added to each well was also calculated using the following formula. The measurement results of the remaining amount of medium and the volume error for each example are shown in Table 1. Remaining medium volume = Bottom area of ​​well (0.3266 cm 2 )×d1 Volume error (%) Medium remaining volume (μL) / 200μL×100

[0058] [Example 3] In a 96-well (8×12) microplate having an embodiment as exemplified in FIG. 1 and FIG. 2, the remaining medium amount and volume error corresponding to the value of the distance d1 shown in Table 2 were calculated in the same manner.

[0059] [Table 1]

[0060] [Table 2]

[0061] As shown in Table 1, Example 1, in which distance d1 is 400 μm or less and distance d2 is 200 μm or less, had a smaller amount of medium remaining and a smaller volume error when applied to an automatic culture device than Example 2, which did not satisfy these conditions. Also, as shown in Table 2, when distance d1 is 400 μm or less, the volume error can be kept to 7% or less. [Explanation of symbols]

[0062] 1 to 3...microplate, 10...upper surface portion, 10a...upper surface, 12...periphery wall portion, 14...well, 16, 16A...bottom surface, 20...frame body, 22...flat base material, 24...tubular portion.

Claims

1. A microplate conforming to the ANSI / SBS standard having a top surface that is rectangular in plan view, a peripheral wall that hangs down vertically from the outer edge of the top surface, and a plurality of cylindrical wells with bottoms each having a circular opening formed on the top surface of the top surface, The plurality of wells are arranged in a rectangular matrix of n×m in plan view, A microplate, wherein the distance between the deepest parts of the two wells that are furthest apart in a direction perpendicular to a reference plane calculated by the reference plane calculation method described below is 400 μm or less. (Reference surface calculation method) Of all the wells in a microplate conforming to the ANSI / SBS standard, which has a rectangular upper surface in plan view, a peripheral wall portion hanging vertically from the outer edge of the upper surface, and a plurality of cylindrical wells with bottoms each having a circular opening formed on the upper surface of the upper surface, four wells are selected that are located at each vertex of the rectangle in plan view and that have the largest area of ​​the rectangle. A three-dimensional orthogonal coordinate system (x, y, z) is set, which has an xy plane passing through the deepest parts of two wells that are diagonally positioned in a planar view among the four wells (however, if the bottom surface of the well is flat, the center of the bottom surface of the well in a planar view), the midpoint of the deepest parts of the two wells is the origin, and the xy coordinates of the deepest parts of the four wells are (-X, Y), (X, Y), (-X, -Y), and (X, -Y), respectively. In the three-dimensional orthogonal coordinate system (x, y, z), when the coordinates of the deepest parts of the four wells are (-X, Y, Z1), (X, Y, Z2), (-X, -Y, Z3), and (X, -Y, Z4), the planes represented by the following formulas 1 to 4 are taken as the reference plane. [0010]

2. A microplate conforming to the ANSI / SBS standard having an upper surface having a rectangular shape in a plan view, a peripheral wall portion vertically hanging down from the outer edge of the upper surface, and a plurality of cylindrical wells with bottoms each having a circular opening formed on the upper surface of the upper surface, the plurality of wells being arranged vertically and horizontally in a rectangular matrix of n x m pieces in a plan view, A microplate in which the distance between a reference plane calculated by the reference plane calculation method described below and the bottom of each well is 200 μm or less. (Reference surface calculation method) Of all the wells in a microplate conforming to the ANSI / SBS standard, which has a rectangular upper surface in plan view, a peripheral wall portion hanging vertically from the outer edge of the upper surface, and a plurality of cylindrical wells with bottoms each having a circular opening formed on the upper surface of the upper surface, four wells are selected that are located at each vertex of the rectangle in plan view and that have the largest area of ​​the rectangle. A three-dimensional orthogonal coordinate system (x, y, z) is set up, in which the xy plane passes through the deepest parts of two wells that are diagonally positioned in a plan view among the four wells (however, if the bottom surface of the well is flat, the center of the bottom surface of the well in a plan view), the midpoint of the deepest parts of the two wells is point 0, and the xy coordinates of the deepest parts of the four wells are (-X, Y), (X, Y), (-X, -Y), and (X, -Y), respectively. In the three-dimensional orthogonal coordinate system (x, y, z), when the coordinates of the deepest parts of the four wells are (-X, Y, Z1), (X, Y, Z2), (-X, -Y, Z3), and (X, -Y, Z4), the planes represented by the following formulas 1 to 4 are taken as the reference plane. [0025]

3. 3. The microplate according to claim 1 or 2, wherein a cross-sectional shape of the bottom surface of the well when cut along a plane parallel to the depth direction passing through the center of the bottom surface in a plan view is a downwardly convex arc shape, a downwardly convex broken line shape, or a straight line shape.

4. The device includes a frame and a flat base material, The microplate according to any one of claims 1 to 3, wherein the frame comprises a plurality of cylindrical portions forming peripheral walls of each of the wells, and the flat substrate is attached to lower end sides of the plurality of cylindrical portions to form the plurality of wells.

5. The microplate according to any one of claims 1 to 4, wherein a plurality of fine wells are formed in the bottom surface of the well.

Images