JP6741112B2 - Cell handling container - Google Patents

Description

Embodiments of the present disclosure relate to a cell handling container, and more particularly to a cell handling container provided with a region forming a drop.
This application applies to Japanese Patent Application No. 2016-195121 filed on September 30, 2016, Japanese Patent Application No. 2017-079258 filed on April 12, 2017 and June 27, 2017. Based on the filed Japanese Patent Application No. 2017-125395, the priority is claimed and the contents thereof are incorporated herein.

In a cell handling container, various effects have been obtained by imparting hydrophilicity to the surface of the container and making the surface hydrophilic. For example, in the case of a container that handles adherent cells, by making the surface of the container a hydrophilic surface, excellent initial adhesiveness and proliferation of cells can be obtained. Further, in the case of a container having microwells for culturing cells such as fertilized eggs, by making the surface of the microwells a hydrophilic surface, bubbles generated in the microwells when the culture solution is added can be easily generated. It can be removed and workability can be improved.

On the other hand, when handling cells such as fertilized eggs, form a thick drop of the culture solution (also called droplets) on the surface of the container, and put the fertilized eggs etc. into this with a pipette for washing and culturing. There are many. However, if the surface of the container is a hydrophilic surface, it is difficult to form the drop, and even if the drop is formed, the drop tends to be crushed.

In order to solve such a problem, for example, in the cell handling container described in Patent Document 1, a plurality of wells are provided at the bottom of the container and a drop is formed in each well to perform washing and culturing work. ..

JP, 2006-280298, A

However, in the cell handling container described in Patent Document 1 described above, when a drop is formed using a pipette to wash or culture cells, the tip of the pipette may come into contact with the edge or side wall of the well and be damaged. There is a nature. The risk of breakage increases, especially if the wells are deep. For this reason, the operator is obliged to work with great care, which causes a new problem that it is difficult to improve workability.

The embodiment of the present disclosure has been made in view of the above points, and an object thereof is to provide a cell handling container that can easily form a drop even on a hydrophilic surface.

A cell handling container according to an embodiment of the present disclosure is a cell handling container, and is provided continuously and intermittently along a circumferential direction of the upper surface and a liquid drop forming region and extends downward. At least one drop forming portion having a side surface is provided, and the side surface extends downward from the edge of the upper surface to a range of 5 μm or more in the vertical direction.

In the cell handling container according to the embodiment of the present disclosure, it is preferable that the minimum width of the upper surface in the horizontal direction is 0.5 mm or more and 10 mm or less.

In the cell handling container according to the embodiment of the present disclosure, it is preferable that the upper surface is a flat surface.

In the cell handling container according to the embodiment of the present disclosure, it is preferable that an angle formed by the upper surface and the side surface is 136° or less at an edge of the upper surface.

In the cell handling container according to an embodiment of the present disclosure, the cell handling container includes a container bottom portion, and between the container bottom portion and the drop forming portion, a groove portion surrounding the drop forming portion is provided, It is preferable that at least a part of the side surface of the drop forming portion constitutes one of the side surfaces of the groove portion.

Further, in the cell handling container according to the embodiment of the present disclosure, in the horizontal direction, the distance from the edge of the upper surface of the drop forming portion to the edge of the upper surface of the container bottom portion that is adjacent via the groove is 0. It is suitable that it is 01 mm or more.

Further, in the cell handling container according to an embodiment of the present disclosure, the cell handling container includes a surrounding wall portion surrounding the drop forming portion, and an upper end surface of the surrounding wall portion is flush with the upper surface of the drop forming portion. Is preferred.

Further, in the cell handling container according to an embodiment of the present disclosure, it is preferable that a distance from an edge of the upper surface of the drop forming portion to an outer edge of the upper end surface of the surrounding wall portion in the horizontal direction is 1 mm or less. is there.

Further, in the cell handling container according to an embodiment of the present disclosure, the cell handling container further comprises a container bottom portion provided around the surrounding wall portion, and between the container bottom portion and the surrounding wall portion, It is preferable that a groove portion surrounding the surrounding wall portion is provided.

Further, in the cell handling container according to an embodiment of the present disclosure, it is preferable that the upper surface of the drop forming portion is flush with the upper surface of the container bottom portion.

Further, in the cell handling container according to the embodiment of the present disclosure, it is preferable that at least the upper surface of the drop forming portion is a hydrophilic surface.

In the cell handling container according to the embodiment of the present disclosure, it is preferable that the hydrophilic surface has a water contact angle of less than 80°.

Further, in the cell handling container according to an embodiment of the present disclosure, it is preferable that the drop forming portions are plural and the upper surfaces of the plurality of drop forming portions are flush with each other.

Furthermore, in the cell handling container according to an embodiment of the present disclosure, the cell handling container further includes a microwell that stores cells, and a liquid storage unit that surrounds the microwell and stores a liquid, and the liquid storage It is preferable that the bottom surface of the portion is flush with the top surface of the drop forming portion.

Furthermore, in the cell handling container according to an embodiment of the present disclosure, the cell handling container further includes a microwell that contains cells, and a liquid storage unit that surrounds the microwell and stores a liquid. It is preferable that the bottom surface of the is flush with the top surface of the drop forming portion.

A cell handling container according to an embodiment of the present disclosure is a cell handling container, and includes a container bottom portion, a drop forming portion that is provided at the container bottom portion, and has a concave portion that opens upward and a peripheral wall portion that forms the concave portion. The upper end portion and the recess of the peripheral wall portion are liquid drop forming regions, and the distance from the upper end portion of the peripheral wall portion to the upper surface of the container bottom portion in the vertical direction is h1, and the peripheral wall portion is When the distance from the upper end to the deepest portion of the recess is h2, h1+h2<0.2 mm.

Further, the cell handling container according to the embodiment of the present disclosure is a cell handling container, and is a drop having a container bottom, a recess provided on the container bottom, opening upward, and a peripheral wall forming the recess. A forming part, and a groove part provided between the container bottom part and the peripheral wall part and surrounding the peripheral wall part, and the upper end part of the peripheral wall part and the concave part are liquid drop forming regions, In the direction, h3+h4≦1 mm, where h3 is the distance from the upper end of the peripheral wall to the upper surface of the container bottom, and h4 is the distance from the upper end of the peripheral wall to the deepest part of the recess. It is characterized by that.

In the cell handling container according to an embodiment of the present disclosure, it is preferable that the outer wall surface of the peripheral wall portion extends downward from the edge of the upper end portion to a range of 5 μm or more in the vertical direction.

In the cell handling container according to an embodiment of the present disclosure, it is preferable that an angle formed by the upper end portion of the peripheral wall portion and the outer wall surface is 136° or less at an edge of the upper end portion.

In the cell handling container according to an embodiment of the present disclosure, it is preferable that the minimum width of the upper end portion of the peripheral wall portion is 0.5 mm or more and 10 mm or less in the horizontal direction.

In the cell handling container according to an embodiment of the present disclosure, it is preferable that the upper end portion of the peripheral wall portion is a flat surface and is flush with the upper surface of the container bottom portion.

Further, in the cell handling container according to an embodiment of the present disclosure, the cell handling container includes an enclosing wall portion surrounding the peripheral wall portion, and the upper end portion of the peripheral wall portion is a flat surface, and is located on the enclosing wall portion. It is suitable to be flush with the end face.

Further, in the cell handling container according to an embodiment of the present disclosure, it is preferable that the distance from the edge of the upper end portion of the peripheral wall portion to the outer edge of the upper end surface of the surrounding wall portion is 1 mm or less in the horizontal direction. is there.

Further, in the cell handling container according to an embodiment of the present disclosure, it is preferable that at least the upper end portion of the peripheral wall portion has a hydrophilic surface.

In the cell handling container according to the embodiment of the present disclosure, it is preferable that the hydrophilic surface has a water contact angle of less than 80°.

Furthermore, in the cell handling container according to the embodiment of the present disclosure, it is preferable that the drop forming portions are plural and the upper ends of the peripheral wall portions of the plural drop forming portions are flush with each other.

Furthermore, in the cell handling container according to an embodiment of the present disclosure, the cell handling container further includes a microwell that stores cells, and a liquid storage portion that surrounds the microwell and stores a liquid, and the liquid storage container. It is preferable that the bottom surface of the portion is flush with the upper end portion of the peripheral wall portion of the drop forming portion.

Furthermore, in the cell handling container according to an embodiment of the present disclosure, the cell handling container further includes a microwell that contains cells, and a liquid storage unit that surrounds the microwell and stores a liquid. It is preferable that the bottom surface of the same is flush with the bottom surface of the recess of the drop forming portion.

A cell handling container according to an embodiment of the present disclosure includes at least one drop forming portion having an upper surface and a side surface that is connected to the upper surface via a curved surface portion and extends downward, Among them, at least the upper surface is a liquid drop formation region, and the radius of curvature of the curved surface portion is less than 250 μm.

According to the present disclosure, drops can be easily formed even on a hydrophilic surface.

It is a top view which shows the cell handling container of 1st Embodiment. It is sectional drawing which follows the AA line of FIG. 1A. It is a fragmentary sectional view showing a modification of a drop formation part. It is a fragmentary sectional view showing a modification of a drop formation part. It is a fragmentary sectional view showing a modification of a drop formation part. It is a top view which shows the cell handling container of 2nd Embodiment. It is sectional drawing which follows the BB line of FIG. 3A. It is a fragmentary sectional view showing a modification of a drop formation part and a groove part. It is a fragmentary sectional view showing a modification of a drop formation part and a groove part. It is a fragmentary sectional view showing a modification of a drop formation part and a groove part. It is a fragmentary sectional view showing a modification of a drop formation part and a groove part. It is a fragmentary sectional view showing a modification of a drop formation part and a groove part. It is a fragmentary sectional view showing a modification of a drop formation part and a groove part. It is a fragmentary sectional view showing a modification of a drop formation part and a groove part. It is a fragmentary sectional view showing a modification of a drop formation part and a groove part. It is a partial top view which shows the cell handling container of 3rd Embodiment. It is sectional drawing which follows CC line|wire of FIG. 7A. It is sectional drawing which follows CC line|wire of FIG. 7A. It is a partial top view which shows the cell handling container of 4th Embodiment. It is sectional drawing which follows the DD line|wire of FIG. 8A. It is sectional drawing which follows the DD line|wire of FIG. 8A. It is a top view which shows the cell handling container of 5th Embodiment. It is sectional drawing which follows the EE line of FIG. 9A. It is a fragmentary sectional view showing a modification of a drop formation part and a crevice. It is a fragmentary sectional view showing a modification of a drop formation part and a crevice. It is a fragmentary sectional view showing a modification of a drop formation part and a crevice. It is a fragmentary sectional view showing a modification of a drop formation part and a crevice. It is a fragmentary sectional view showing a modification of a drop formation part and a crevice. It is a fragmentary sectional view showing a modification of a drop formation part and a crevice. It is a fragmentary sectional view showing a modification of a drop formation part and a crevice. It is a fragmentary sectional view showing a modification of a drop formation part and a crevice. It is a fragmentary sectional view showing a modification of a drop formation part. It is a fragmentary sectional view showing a modification of a drop formation part. It is a top view which shows the cell handling container of 6th Embodiment. It is sectional drawing which follows the FF line of FIG. 15A. It is a fragmentary sectional view showing a modification of a peripheral wall part and a crevice. It is a fragmentary sectional view showing a modification of a peripheral wall part and a crevice. It is a fragmentary sectional view showing a modification of a peripheral wall part and a crevice. It is a fragmentary sectional view showing a modification of a peripheral wall part and a crevice. It is a fragmentary sectional view showing a modification of a peripheral wall part. It is a fragmentary sectional view showing a modification of a peripheral wall part. It is a partial top view which shows the cell handling container of 7th Embodiment. It is sectional drawing which follows the GG line of FIG. 19A. It is sectional drawing which follows the GG line of FIG. 19A. It is a partial top view which shows the cell handling container of 8th Embodiment. It is sectional drawing which follows the HH line of FIG. 20A. It is sectional drawing which follows the HH line of FIG. 20A. It is a top view which shows the cell handling container (state which removed the cover body) of 9th Embodiment. It is sectional drawing which follows the II line|wire of FIG. 21A. It is sectional drawing which follows the JJ line of FIG. 21A. It is sectional drawing which follows the KK line of FIG. 21A. It is sectional drawing which follows the LL line of FIG. 21A. It is a top view which shows the cell handling container of 10th Embodiment. It is sectional drawing which follows the MM line of FIG. 22A. It is a top view which shows the cell handling container of 11th Embodiment. It is sectional drawing which follows the NN line of FIG. 23A. It is a perspective view which shows the cell handling container (state which removed the cover body) of 12th Embodiment. It is a top view which shows the container main body of a cell handling container. It is sectional drawing which follows the P-P line of FIG. It is a bottom view which shows the container main body of a cell handling container. It is sectional drawing which follows the QQ line of FIG. 27A. It is a top view which shows the cell handling container of 13th Embodiment. It is sectional drawing which follows the RR line of FIG. 28A. It is an expanded sectional view of a drop formation part. It is an expanded sectional view showing a modification of a drop formation part. 9 is a photograph showing the results of Example 8. 16 is a photograph showing the result of Example 13. FIG. 6 is a schematic partial cross-sectional view showing a drop formed by the drop forming portion of the first embodiment. FIG. 6 is a schematic partial cross-sectional view showing a relationship between an angle α and a water contact angle θ. 3 is a photograph showing the results of some samples according to Example 1. FIG. 28 is a diagram showing the shape of the drop forming portion of Example 17; FIG. 16 is a diagram showing the shape of a drop forming portion of Example 18. 20 is a diagram showing conditions and results of Example 19. FIG. It is a figure which shows the conditions and result of Example 20. It is a top view which shows the cell handling container of 14th Embodiment. It is sectional drawing which follows the BB line of FIG. 37A. 20 is a photograph of the cell handling container used in Example 21. 20 is a photograph of the cell handling container used in Example 21. 20 is a photograph of the cell handling container used in Example 21. FIG. 13 is a diagram showing the shape of a cell handling container used in Example 21. FIG. 20 is a schematic diagram showing a change in drop shape before vibration in Example 21. FIG. 20 is a schematic diagram showing a change in drop shape after vibration in Example 21.

Hereinafter, an embodiment of a cell handling container according to the present invention will be described with reference to the drawings. In the description of the drawings, the same elements will be denoted by the same reference symbols, without redundant description. Moreover, in the following description, the up/down, left/right, and front/rear positions and directions are the positions and directions in the normal usage state of the cell handling container. Further, in each of the drawings, the distances and intervals between the respective constituent parts may be drawn larger or smaller than the dimensions described in the embodiments in order to facilitate understanding of the invention.

<First Embodiment>
FIG. 1A is a plan view showing the cell handling container of the first embodiment, and FIG. 1B is a sectional view taken along the line AA of FIG. 1A. The cell handling container 1 of the present embodiment is a container used for handling such as washing, culturing, temporary storage of cells including fertilized eggs, egg cells, ES cells (embryonic stem cells) and iPS cells (artificial pluripotent stem cells). Is.

Here, the egg cell refers to an unfertilized egg cell, and includes an immature oocyte and a mature oocyte. After fertilization, the number of cells of the fertilized egg increases at the 2-cell stage, 4-cell stage, and 8-cell stage due to cleavage, and develops into blastocysts through the morula. Fertilized eggs include early embryos such as 2-cell embryos, 4-cell embryos and 8-cell embryos, morula, blastocysts (including early blastocysts, expanded blastocysts and escape blastocysts). Blastocyst refers to an embryo that consists of an outer cell that has the potential to form a placenta and an inner cell mass that has the potential to form an embryo. ES cells refer to undifferentiated pluripotent or totipotent cells obtained from the inner cell mass of blastocysts. The iPS cells refer to cells that have pluripotency similar to ES cells by introducing several kinds of genes (transcription factors) into somatic cells (mainly fibroblasts).

In addition, the cell handling container 1 of the present embodiment is preferably suitable for culturing mammalian cells and avian cells, particularly mammalian cells. Mammals refer to warm-blooded vertebrates, for example, primates such as humans and monkeys, rodents such as mice, rats and rabbits, companion animals such as dogs and cats, and livestock such as cows, horses and pigs. Is mentioned. The cell handling container 1 of the present embodiment is suitable for culturing human or bovine fertilized eggs, and particularly suitable for culturing human fertilized eggs.

As shown in FIGS. 1A and 1B, the cell handling container 1 includes a bottomed cylindrical container body 1a having an upper opening, and a lid body 1b detachably installed on the container body 1a. The lid 1b has a disk-shaped top plate portion 51 that closes the opening of the container body 1a, and a cylindrical peripheral wall portion 52 that extends downward from the peripheral edge of the top plate portion 51. On the other hand, the container body 1a is, for example, a dish type having an inner diameter of 35 mm or 60 mm, and has a substantially disc-shaped container bottom 10 having a top surface 10a and a bottom surface 10b arranged in parallel with each other, and a peripheral edge of the container bottom portion 10. It has a cylindrical side wall portion 11 standing upright and a cell containing area 12 provided at a central position of the container bottom portion 10.

The cell storage area 12 includes a plurality of microwells 13 for storing and culturing cells such as fertilized eggs, and a cylindrical culture solution storage section (liquid storage section) 14 that surrounds the microwells 13 and stores a culture solution. Have. Each of the microwells 13 has a U-shaped cross-section with an upward opening, and is arranged in the front, rear, left, and right directions in close proximity to each other at regular intervals. Note that the cross section of the microwell 13 does not necessarily have to be a U-shape with an upward opening, and may have, for example, a truncated cone shape with an upward-opening cross section, or a U-shaped cross section having an arc portion, or another shape. good. Further, the bottom surface of the microwell 13 may be a flat surface or a tapered surface that is inclined from the outside toward the center.

In addition, the upper surface 10a of the container bottom 10 is provided with a plurality of (four in this embodiment) drop forming portions 15 that project upward from the upper surface 10a to form liquid drops such as a culture solution. These drop forming portions 15 each have a cylindrical shape, are integrally formed with the container bottom portion 10, and are arranged at equal intervals around the cell containing area 12 so as to surround the cell containing area 12.

The drop forming portion 15 has an upper surface 15a serving as a drop forming region and a side surface 15b extending downward along the entire circumference of the upper surface 15a. The upper surface 15a is a flat surface. By making the upper surface 15a flat as described above, when cells such as fertilized eggs arranged on the upper surface 15a of the drop forming portion 15 are observed through a microscope with a microscope, light reflection and scattering are less likely to occur, resulting in a clear image. It is possible to obtain a clear observation image. The flat surface here is not limited to a completely smooth horizontal surface, but may be a substantially horizontal surface, such as a flat portion of a general injection-molded product having a difference of about several μm. It means that there may be a manufacturing surface roughness of about 1% or less, which is within the standard tolerance according to JIS B 0419-991.

The side surface 15b is provided over the entire circumference along the circumferential direction of the upper surface 15a and is arranged at a right angle to the upper surface 15a. Therefore, the angle α formed by the upper surface 15a and the side surface 15b is 90° at the edge P1 of the upper surface. The side surface 15b extends from the edge P1 of the upper surface 15a to a range of 5 μm or more.

In the present embodiment, since the drop forming portion 15 is erected from the upper surface 10a of the container bottom portion 10 and the upper surface 15a is a flat surface, the extending range of the side surface 15b in the vertical direction is the upper surface 10a of the container bottom portion 10. It is the same as the height t of the upper surface 15a of the drop forming portion 15. In other words, the height t of the upper surface 15a of the drop forming portion 15 with respect to the upper surface 10a of the container bottom portion 10 is 5 μm or more. The lower limit value of the height t, 5 μm, is set based on the condition that the drop can be formed by the surface tension of the culture solution when the drop is formed using the culture solution of the fertilized egg. Further, the height t of the upper surface 15a of the drop forming portion 15 with respect to the upper surface 10a of the container bottom 10 is preferably 10 μm or more, more preferably 50 μm or more, further preferably 100 μm or more, and further preferably 250 μm or more. Is.

These lower limit values are because the higher the height t of the upper surface 15a of the drop forming portion 15 with respect to the upper surface 10a of the container bottom 10 is, the easier the formed drops are held, and when the height t is increased to a certain level, sufficient holding is obtained. is there. For example, if the height t is 50 μm or more, a thick drop can be created even if the liquid amount is increased, and if the height t is 100 μm or more, a thick drop can be stably created even if the liquid amount is increased. Furthermore, if the thickness is 250 μm or more, a thick drop can be formed more stably even if the liquid amount is increased. The upper limit of the height t is not particularly limited as long as the upper surface 15a of the drop forming portion 15 does not interfere with the top plate portion 51 of the lid 1b when closing the lid 1b.

In the horizontal direction, the width of the upper surface 15a (here, the diameter D of the upper surface 15a) is 0.5 mm or more and 10 mm or less. Preferably, the diameter D of the upper surface 15a is 3 mm or more and 6 mm or less. A fertilized egg may grow up to a diameter of about 250 μm in a blastocyst and up to a diameter of about 500 μm including the zona pellucida. The diameter D of the upper surface 15a is set to be 0.5 mm or more, which is the maximum diameter of the fertilized egg, in order to facilitate the culturing or washing work while considering the size of the fertilized egg. It should be noted that cells such as fertilized eggs may have a spherical shape or an elliptical shape due to growth. Therefore, the cell diameter in the present embodiment means the diameter when it is spherical, and the maximum width when it is not spherical.

On the other hand, the width of the upper surface 15a (that is, the diameter D of the upper surface 15a) may be set to 3.4 mm or more and 7.2 mm or less according to the following description, and is set to 3.8 mm or more and 5.4 mm or less. preferable. As shown in FIG. 31A, when the end portion of the drop S formed by the drop forming portion 15 rises vertically, that is, the boundary point between the drop S and the upper surface 15a of the drop forming portion 15 (here, the upper surface 15a described later). When the angle formed by the upper surface 15a and the drop (that is, the water contact angle θ) at the edge P1) of the drop forming portion 15 is 90°, the upper surface 15a of the drop forming portion 15 has a circular shape with a radius r′ [mm] in a plan view (r '=D/2), the thickness h'[mm] of the drop formed on the upper surface 15a satisfies the relational expression of h'=3V/(2πr' ² ). Here, when h'exceeds r', the drop is likely to collapse, so that h'≤r', and V is the culture solution volume [μ
L] and π indicate the circular constant, respectively.

The volume V of the culture solution is preferably 10 μL or more and 100 μL or less, more preferably 15 μL or more and 40 μL or less when culturing or washing cells such as fertilized eggs. Further, as h'is larger, it becomes easier to operate a device such as a pipette, but when h'exceeds r', the drop is likely to collapse. Therefore, the workability is good when the drop thickness h'is h'=r'.

Based on the above equation, when V=10 μL and h′=r′, r′=1.7 mm (that is, D=3.4 mm). On the other hand, when V=100 μL and h′=r′, r′=3.6 mm (that is, D=7.2 mm). Similarly, when V=15 μL and h′=r′, r′=1.9 mm (that is, D=3.8 mm). When V=40 μL and h′=r′, r′=2.7 mm (that is, D=5.4 mm).

From the above, while ensuring good operability of instruments such as pipettes, when the thickness h'of the formed drop is the same as the radius r'of the upper surface 15a (h'=r'), the culture solution volume V is 10 μL. When it is 100 μL or less, the diameter D is in the range of 3.4 mm or more and 7.2 mm or less, and when the culture solution volume V is 15 μL or more and 40 μL or less, the diameter D is 3.8 mm or more and 5.4 mm or less. It becomes the range.

Further, the width of the upper surface 15a (that is, the diameter D of the upper surface 15a) may be set to 2.0 mm or more by the following description. Specifically, irrespective of the culture solution volume V, when h′=r′ and h′≧1 mm, r′≧1 mm (that is, D≧2 mm). Even in this case, the operability of instruments such as pipettes is facilitated. As a result, while ensuring good operability of instruments such as pipettes, when the thickness h′ of the formed drop is the same as the radius r′ of the upper surface 15a (h′=r′), when h′≧1 mm Moreover, the diameter D becomes 2 mm or more.

Although the upper surface 15a of the drop forming portion 15 is circular in a plan view in the present embodiment, it may be polygonal such as quadrangle, elliptical, or rounded square. However, considering the ease of holding the formed drops, it is preferable that the upper surface 15a is circular.

On the other hand, when the diameter D of the upper surface 15a exceeds 10 mm, the formed drop is too large with respect to the size of the fertilized egg, which makes it difficult to find the fertilized egg during washing or culturing the fertilized egg, and is also limited. The number of drop forming parts that can be installed in the space inside the container is also reduced. Considering that a fertilized egg can be smoothly found to efficiently perform washing and culturing work, and that more drop forming portions can be installed in a limited space, the diameter D of the upper surface 15a may be 10 mm or less. preferable.

Although four drop forming portions 15 are provided in the present embodiment, the upper surfaces 15a of these drop forming portions 15 are all flush with each other. By doing so, for example, when the fertilized egg is washed a plurality of times with an instrument such as a pipette, the instrument can be slid and moved, so that workability can be further enhanced.

The material of the cell handling container 1 is not particularly limited. Specifically, inorganic materials such as metal, glass, and silicon, plastics such as polystyrene resin, polyester resin, polyethylene resin, polypropylene resin, ABS resin, nylon, acrylic resin, fluororesin,
Organic materials represented by polycarbonate resin, polyurethane resin, methylpentene resin, phenol resin, melamine resin, epoxy resin and vinyl chloride resin can be mentioned. The cell handling container 1 can be manufactured by a method known to those skilled in the art. For example, when using a plastic material, it can be manufactured by injection molding.

In the cell handling container 1 configured as described above, the drop forming portion 15 projects upward from the upper surface 10a of the container bottom portion 10, and the upper surface 15a of the drop forming portion 15 is not directly connected to the upper surface 10a of the container bottom portion 10. Further, the upper surface 15a is a flat surface, and the height t of the container bottom portion 10 with respect to the upper surface 10a is 5 μm or more. Therefore, when a drop of the culture solution is formed on the upper surface 15a, the drop can be easily formed by the surface tension of the culture solution. As a result, even if the upper surface 15a of the drop forming portion 15 is a hydrophilic surface, a thick drop S can be easily formed on the upper surface 15a as shown by the chain double-dashed line in FIG. 1B. Then, cells C such as fertilized eggs can be placed in the drop S.

Moreover, since the upper surface 15a, which is the drop forming region, is higher than the upper surface 10a of the container bottom portion 10, the tip of an instrument such as a pipette is located at the edge or side wall of the well as compared with the conventional case where a drop forming well is provided. It is possible to improve workability without worrying about contact with and damage.

In addition, in the present embodiment, in order to improve the workability by easily removing the bubbles in the microwell 13 when the culture solution is added, the surface of the microwell 13 and the surface of the cell containing area 12 are made hydrophilic. Good as the surface. In order to make these surfaces hydrophilic, a hydrophilic material may be used for the entire cell handling container 1 including the cell accommodation area 12 and the drop forming portion 15, or these surfaces may be subjected to a hydrophilic treatment. May be. The hydrophilic treatment can be patterned by using a mask, but the treatment on the entire surface is preferable from the viewpoint of manufacturing cost and quality control.

The hydrophilization treatment can be performed by a method commonly used in the art, and is not particularly limited. Examples include plasma treatment, coating treatment, UV irradiation treatment, EB irradiation treatment, and graft polymerization treatment of hydrophilic polymer on the surface. However, the shape of the object to be treated has a three-dimensionally fine and complicated structure. However, the plasma treatment is preferable from the viewpoint that the whole can be uniformly treated. The hydrophilic treatment is preferably performed after the cell handling container 1 is molded. The water contact angle θ on the hydrophilic surface is preferably less than 80° (see FIG. 1B).

Normally, when the water contact angle θ is less than 80°, when attempting to form a drop on a flat surface, the drop will be crushed, and even if a drop can be formed, it is very difficult to operate a device such as a pipette. There is a problem that cells such as eggs cannot be washed or cultured. In particular, when the water contact angle θ is 60° or less, the above problem becomes more remarkable. For example, when culturing or washing cells such as fertilized eggs, the amount of culture solution is used in the range of 15 μL or more and 30 μL or less, and from the viewpoint of ensuring good operability of instruments such as pipettes, the thickness of the formed drop is It is preferably 1 mm or more, more preferably 1.5 mm or more. However, on a flat surface having a water contact angle θ of less than 80° in the range of the culture solution volume of 15 μL or more and 30 μL or less, the thickness of the drop is 1.5 mm or less, which makes it difficult to operate the device. On a flat surface having a water contact angle θ of 60° or less, the thickness of the drop is 1.0 mm or less, which makes it very difficult to operate the device.

On the other hand, according to the drop forming portion 15 of the present embodiment, a thick drop can be easily formed even when the water contact angle θ is less than 80°, and a thick drop can be formed even when the water contact angle θ is 60° or less. It can be easily formed, and a thick drop can be easily formed even at 10° or less.

Various modifications can be considered for the shape of the drop forming portion 15 according to the present embodiment.
For example, in the modification shown in FIG. 2A, the side surface 15c of the drop forming portion 15 is an inclined surface that inclines outward as it goes downward in a cross-sectional view. For this reason, the cross section of the drop forming portion 15 has a truncated cone shape. At this time, at the edge P1 of the upper surface 15a, the angle α formed by the upper surface 15a and the side surface 15c becomes an obtuse angle.

Then, considering the influence of the difference in angle on the surface tension of the culture medium, the angle α is preferably 136° or less. As shown in FIG. 31B, at the edge P1 of the upper surface 15a, the relational expression of angle α=180°−θ is satisfied. θ is a water contact angle, and is calculated by θ=2 arctan (h′/r′) from the θ/2 method generally used in water contact angle measurement, where h′ is the thickness of the drop and r′ is It is the radius (r'=D/2) of the upper surface 15a of the drop forming portion 15. For example, when a drop is formed by a drop forming unit with r′=2.5 mm (that is, D=5 mm), if the angle α is 136° or less, h′ is 1 mm or more (it is easy to operate a device such as a pipette). Height). Further, the smaller the angle α, the more easily the formed drop is formed, and the formed drop is easily held, so that the angle α is more preferably 100° or less, further preferably 95° or less, and further preferably 93° or less. It is more preferably 92° or less, still more preferably 91° or less. In addition, the angle α here includes one whose tip is not sharp due to manufacturing problems. Further, in this case, when the water contact angle θ′ of the entire inner surface of the container excluding the edge P1 is θ′≧α-90° (where α≧90°), the water contact angle of the drop forming portion at the edge P1. Drops can be formed when θ is in the range of θ≦90°. For example, in the drop forming portion of α=120°, when the water contact angle θ′ of the entire inner surface of the container excluding the edge P1 is θ′≧30°, and the range is θ≦90°, a drop is formed. It is possible.

In the modification shown in FIG. 2B, the side surface 15d is a step surface having a vertical portion and an inclined portion in order from above in a cross-sectional view. The vertical portion is a portion extending at a right angle to the upper surface 15a, and the inclined portion is a portion inclined outward as it goes downward. At this time, at the edge P1 of the upper surface, the angle α formed by the upper surface 15a and the vertical portion of the side surface 15d is 90°.

In the modification shown in FIG. 2C, the corner between the side surface 15e of the drop forming portion 15 and the upper surface 10a of the container bottom portion 10 is processed to have a curved portion in a sectional view. Therefore, the side surface 15e has a vertical portion and a curved portion in order from above. At this time, at the edge P1 of the upper surface 15a, the angle α formed by the upper surface 15a and the vertical portion of the side surface 15e is 90°.

<Second Embodiment>
3A is a plan view showing the cell handling container of the second embodiment, and FIG. 3B is a sectional view taken along the line BB of FIG. 3A. The difference between the cell handling container 2 of the present embodiment and the first embodiment is that a groove portion 17 is provided between the drop forming portion 16 and the container bottom portion 10, and the upper surface 16a of the drop forming portion 16 is of the container bottom portion 10. It is flush with the upper surface 10a. Since other configurations are similar to those of the first embodiment, duplicated description will be omitted.

Specifically, the container bottom 10 is provided with a plurality of drop forming portions 16 each having an upper surface 16a serving as a drop forming area and a side surface 16b continuously provided along the circumferential direction of the upper surface 16a and extending downward (here Then, four) are provided. The drop forming portion 16 is formed in a cylindrical shape like the drop forming portion 15 of the first embodiment. Further, the upper surface 16 a of the drop forming portion 16 is a flat surface and is flush with the upper surface 10 a of the container bottom portion 10. Furthermore, as shown in FIG. 3B, the upper surface 16 a of the drop forming portion 16 is flush with the bottom surface 14 a of the culture fluid storage portion 14 in the cell storage area 12.

The side surface 16b is provided over the entire circumference of the upper surface 16a, is disposed at a right angle to the upper surface 16a, and extends downward from the edge P1 of the upper surface 16a to a range of 5 μm or more.
In other words, the height t of the upper surface 16a of the drop forming portion 16 with respect to the bottom surface of the groove portion 17 (described later) is 5 μm or more. Here, since the side surface 16b is arranged at a right angle to the upper surface 16a, the angle α formed by the upper surface 16a and the side surface 16b is 90° at the edge P1 of the upper surface 16a. Further, the height t of the upper surface 16a of the drop forming portion 16 with respect to the bottom surface of the groove portion 17 is preferably 10 μm or more, more preferably 50 μm or more, further preferably 100 μm or more, and further preferably 250 μm or more.

These lower limit values become easier to hold the formed drop as the height t of the upper surface 16a of the drop forming portion 16 with respect to the bottom surface of the groove portion 17 (in other words, the deeper the groove portion 17) becomes, and become higher to a certain level. This is because sufficient retention can be obtained. For example, if the height t is 50 μm or more, a thick drop can be created even if the liquid amount is increased, and if the height t is 100 μm or more, a thick drop can be stably created even if the liquid amount is increased. Furthermore, if the thickness is 250 μm or more, a thick drop can be formed more stably even if the liquid amount is increased. On the other hand, as the height t is higher (the groove portion 17 is deeper), the thickness of the bottom surface of the container is smaller, so that the bottom surface of the container is likely to be damaged and is difficult to form from the viewpoint of manufacturing. For example, the bottom thickness of a general cell culture container is often 1 mm, and in that case, if the height t (depth of the groove) is 1 mm, the bottom surface of the container will penetrate. Therefore, the height t (depth of the groove) is preferably less than 1 mm, and more preferably 500 μm or less, further preferably 350 μm or less, further preferably 250 μm or less, and 100 μm or less from the viewpoint of ease of manufacturing by injection molding. Is even more preferable. When the height t (depth of the groove) is 500 μm or less, the possibility that the bottom surface of the container is penetrated or damaged due to physical damage is extremely low.

An annular groove 17 that surrounds the drop forming portion 16 is provided between the drop forming portion 16 and the container bottom portion 10 for each drop forming portion 16. The groove portion 17 is formed in a U-shape whose cross section opens upward. The side surface 16 b of the drop forming portion 16 constitutes one of the side surfaces of the groove portion 17.

In the horizontal direction, the distance d from the edge P1 of the upper surface 16a of the drop forming portion 16 to the edge P2 of the upper surface 10a of the container bottom portion 10 that is adjacent via the groove portion 17 is 0.01 mm or more, preferably 0.05 mm or more. And more preferably 0.1 mm or more, and further preferably 0.4 mm or more. This is because the formed drop is easier to hold as the distance d increases, and sufficient hold is obtained when the distance d increases to a certain level. On the other hand, if the distance d is too large, the number of drop forming portions that can be installed in the limited space decreases, so it is preferably 4 mm or less. In this embodiment, the distance d is 0.5 mm.

In the cell handling container 2 configured as described above, the upper surface 16a of the drop forming portion 16 is separated from the upper surface 10a of the container bottom portion 10 by the groove portion 17, and the upper surface 16a is a flat surface and the side surface 16b is 5 μm or more from the upper surface 16a. Extends down to the range. Therefore, like the first embodiment described above, even if the upper surface 16a is a hydrophilic surface, the drop S of the culture solution can be easily formed on the upper surface 16a. Further, unlike the prior art, there is no concern that the tip of the instrument will come into contact with the edge or side wall of the well and be damaged, so that workability can be improved.

Moreover, since the upper surface 16a of the drop forming portion 16 is flush with the upper surface 10a of the container bottom portion 10, when the fertilized egg is washed a plurality of times with an instrument such as a pipette, the instrument can be slid and moved. Therefore, the workability can be further enhanced.

In addition, in the present embodiment, the bottom surface of the microwell 13 may be flush with the top surface 16 a of the drop forming portion 16. By doing so, for example, when cells such as a fertilized egg are moved between the microwell 13 and the drop forming section 16, or when they are arranged at both locations, they can be observed at the same focus position in the microscope observation, The property can be further improved. Further, the groove portion 17 according to the present embodiment has an annular shape surrounding the drop forming portion 16, but as long as the drop S of the culture solution can be easily formed on the upper surface 16a of the container bottom portion 10, the annular portion has an annular shape. May be partially cut off and may have a substantially C-shape in a top view.

Various modifications can be considered for the shapes of the drop forming portion and the groove portion according to the present embodiment. For example, in the modification shown in FIG. 4A, the side surface 16c of the drop forming portion 16 is an inclined surface that inclines outward as it goes downward in a cross-sectional view. Therefore, the cross section of the drop forming portion 16 has a truncated cone shape. At this time, at the edge P1 of the upper surface 16a, the angle α formed by the upper surface 16a and the side surface 16c is an obtuse angle, but is preferably 136° or less for the above reason. Furthermore, the smaller the angle α, the more easily the formed drop is formed, and the formed drop is easily held. Therefore, the angle α is more preferably 100° or less, further preferably 95° or less, and further preferably 93° or less, It is more preferably 92° or less, still more preferably 91° or less.

In the modification shown in FIG. 4B, the side surface 16d of the drop forming portion 16 is an inclined surface that inclines outward as it goes downward in a cross-sectional view. Therefore, the cross section of the drop forming portion 16 has a truncated cone shape. At this time, at the edge P1 of the upper surface 16a, the angle α formed by the upper surface 16a and the side surface 16d is an obtuse angle, but is preferably 136° or less, more preferably 100° or less, further preferably 95° or less, and 93° or less. More preferably, 92° or less is even more preferable, and 91° or less is even more preferable. At this time, the cross section of the groove portion 19 is V-shaped.

In the modification shown in FIG. 4C, the side surface 16e is a step surface having a vertical portion and an inclined portion in order from above in a cross-sectional view. The vertical portion is a portion that extends at a right angle to the upper surface 16a, and the inclined portion is a portion that is inclined outward as it goes downward. At this time, at the edge P1 of the upper surface 16a, the angle α formed by the upper surface 16a and the vertical portion of the side surface 16e is 90°. The cross section of the groove 20 has a shape in which an upper rectangular shape and a lower inverted triangle are combined.

In the modified example shown in FIG. 5A, the corner portion between the side surface 16f of the drop forming portion 16 and the container bottom portion 10 is processed to have a curved portion in a cross-sectional view. Therefore, the side surface 16f has a vertical portion and a curved portion in order from above. At this time, at the edge P1 of the upper surface 16a, the angle α formed by the upper surface 16a and the vertical portion of the side surface 16f is 90°. The groove 21 has a U-shaped cross section.

In the modification shown in FIG. 5B, the side surface 16g of the drop forming portion 16 is an inclined surface that inclines inward as it goes downward in a cross-sectional view. Therefore, the cross section of the drop forming portion 16 has an inverted truncated cone shape. At this time, at the edge P1 of the upper surface 16a, the angle α formed by the upper surface 16a and the side surface 16g becomes an acute angle. The cross section of the groove 22 has a truncated cone shape that opens upward.

In the modification shown in FIG. 5C, the side surface 16h of the drop forming portion 16 is a step surface having an inclined portion and a vertical portion in order from above in a cross-sectional view. The inclined portion is a portion that is inclined inward as it goes downward, and the vertical portion is a portion that extends at a right angle to the upper surface 16a. At this time, at the edge P1 of the upper surface 16a, the angle α formed by the upper surface 16a and the inclined portion of the side surface 16h becomes an acute angle. The cross section of the groove 23 is a combination of an upper truncated cone shape and a lower rectangular shape.

In the present embodiment, the upper surface 16a of the drop forming portion 16 is flush with the upper surface 10a of the container bottom portion 10. However, as shown in FIG. 6A, the upper surface 16a of the drop forming portion 16 is higher than the upper surface 10a of the container bottom portion 10. You may arrange. In this case, a part of the side surface 16i of the drop forming portion 16 (that is, a portion of the side surface 16i below the upper surface 10a of the container bottom 10) constitutes one side surface of the groove portion 24.

Further, as shown in FIG. 6B, the upper surface 16a of the drop forming portion 16 may be arranged lower than the upper surface 10a of the container bottom portion 10. In this case, the side surface 16j of the drop forming portion 16 constitutes one side surface of the groove portion 25.

<Third Embodiment>
FIG. 7A is a partial plan view showing the cell handling container of the third embodiment, and FIGS. 7B and 7C are cross-sectional views taken along the line CC of FIG. 7A. The difference between the cell handling container 3 of the present embodiment and the second embodiment is that a further surrounding wall portion 26 surrounding the drop forming portion 16 is further provided, and a groove portion 27 is provided between the container bottom portion 10 and the surrounding wall portion 26. Is. Since other configurations are the same as those in the second embodiment, duplicate description will be omitted.

Specifically, an annular surrounding wall portion 26 is provided on the outer periphery of the drop forming portion 16 so as to surround the drop forming portion 16. The surrounding wall portion 26 is arranged at a predetermined distance from the side surface 16b of the drop forming portion 16. The upper end surface 26a of the surrounding wall portion 26 is flush with the upper surface 16a of the drop forming portion 16 and the upper surface 10a of the container bottom portion 10. In the horizontal direction, the distance c from the edge P1 of the upper surface 16a of the drop forming portion 16 to the outer edge P3 of the upper end surface 26a of the surrounding wall portion 26 is 1 mm or less, preferably 0.8 mm or less, and 0.5 mm or less. Is more preferable.

A groove 27 that surrounds the surrounding wall 26 is provided between the surrounding wall 26 and the container bottom 10. The groove portion 27 of the present embodiment is formed in a U-shape with its cross section opening upward, similarly to the groove portion 17 of the above-described second embodiment. Then, one of the side surfaces of the groove portion 27 is formed by the side surface of the surrounding wall portion 26. As for the cross-sectional shape of the groove 27, the modified examples of the groove of the second embodiment (see FIGS. 4A to 4C and 5A to 5C) are also applied.

Further, in the horizontal direction, the distance d1 from the edge P1 of the upper surface 16a of the drop forming portion 16 to the closest edge P4 of the upper end surface 26a of the surrounding wall portion 26 and the groove 27 from the outer edge P3 of the upper end surface 26a of the surrounding wall portion 26 are defined. The distance d2 to the edge P2 of the upper surface 10a of the container bottom 10 adjacent to each other is 0.01 mm or more, preferably 0.05 mm or more, more preferably 0.1 mm or more, and further preferably It is 0.4 mm or more. This is because the formed drop is easier to hold as the distances d1 and d2 increase, and sufficient hold can be obtained when the distance d1 and d2 increase to a certain level. On the other hand, if the distances d1 and d2 are too large, the number of drop forming portions that can be installed in the limited space decreases, so it is preferable that each is 4 mm or less.

In the cell handling container 3 configured in this manner, the upper surface 16a of the drop forming portion 16 is isolated from the surroundings by the space between the drop forming portion 16 and the surrounding wall portion 26. Therefore, as in the second embodiment described above, the upper surface 16a is Even if it is a hydrophilic surface, the drop S of the culture solution can be easily formed on the upper surface 16a, and there is no concern that the tip of the instrument will contact the edge or side wall of the well and be damaged as in the conventional case. Workability can be improved.

Further, the surrounding wall portion 26 surrounding the drop forming portion 16 is provided, and the distance c from the edge P1 of the upper surface 16a to the outer edge P3 of the upper end surface 26a of the surrounding wall portion 26 is 1 mm or less, and the surrounding wall portion 26 and the container bottom portion 10 are provided. Since the groove section 27 is provided between the drop forming section 16 and the upper surface 16a of the drop forming section 16, when the drop S spreads to the surroundings due to vibration or the like, the drop S forms the surrounding wall section 26.
It is possible to extend to the outer edge P3 of the surrounding wall portion 26 (see FIG. 7C). As a result, it is possible to prevent the drop S from collapsing. In this case, although the thickness of the drop S is reduced by spreading to the outer edge P3, it does not affect the culture or washing of the fertilized egg. Although the upper end surface 26a of the surrounding wall portion 26 and the upper surface 16a of the drop forming portion 16 are flush with each other in the present embodiment, they may not be flush with each other.

<Fourth Embodiment>
FIG. 8A is a partial plan view showing the cell handling container of the fourth embodiment, and FIGS. 8B and 8C are sectional views taken along the line D-D of FIG. 8A. The difference between the cell handling container 4 of the present embodiment and the second embodiment is that the side surface 16k of the drop forming portion 16 is formed by two or more stepped surfaces. Since other configurations are the same as those in the second embodiment, duplicate description will be omitted.

Specifically, the side surface 16k is formed in a stepped shape (here, three steps) so as to extend downward along the entire circumference of the upper surface 16a and to expand from the inner side to the outer side. The side surface 16k is, in order from above, a first portion 16k1 arranged at a right angle to the upper surface 16a, a second portion 16k2 having a horizontal portion and a vertical portion connected to the first portion 16k1, and a second portion 16k2. And a third portion 16k3 having a horizontal portion and a vertical portion.

In the vertical direction, the distance from the edge P1 of the upper surface 16a of the side surface 16k to the horizontal portion of the second portion 16k2 (that is, the height of the first portion 16k1) t1, the horizontal portion of the second portion 16k2 to the horizontal portion of the third portion 16k3. Distance to the portion (that is, the height of the second portion 16k2) t2, and distance from the horizontal portion of the third portion 16k3 to the bottom surface of the groove portion 28 (described later) (that is, the height of the third portion 16k3) t3 Are 5 μm or more, respectively. The heights t1, t2 and t3 are preferably 10 μm or more, more preferably 50 μm or more, further preferably 100 μm or more, and further preferably 250 μm or more.

These lower limit values are because the higher the above-mentioned heights t1, t2, and t3, the easier it is for the formed drops to be retained, and for the lower limits to reach a certain level, sufficient retention can be obtained. For example, when the heights t1, t2, and t3 are 50 μm or more, a thick drop can be created even when the liquid amount is increased, and when the heights t1, t2, and t3 are 100 μm or more, a thick drop is generated even if the liquid amount is increased. It is possible to stably create, and if it is 250 μm or more, it is possible to more stably create a thick drop even if the liquid amount is increased. The upper limits of the heights t1, t2, and t3 are not particularly limited, but the higher the height, the more difficult it is to form from the viewpoint of manufacturing.

At this time, since the first portion 16k1 of the side surface 16k is arranged at a right angle to the upper surface 16a, the angle α formed by the upper surface 16a and the first portion 16k1 of the side surface 16k at the edge P1 of the upper surface 16a is 90°. is there.

Further, an annular groove portion 28 surrounding the drop forming portion 16 is provided between the drop forming portion 16 and the container bottom portion 10. In cross-sectional view, the groove portion 28 opens upward, one of the side surfaces of the groove portion 28 is formed stepwise by the side surface 16k of the drop forming portion 16, and the other is a vertical surface.

In the horizontal direction, the distance (that is, the width of the second portion 16k2) y1 from the edge P1 of the upper surface 16a of the drop forming portion 16 to the vertical portion of the second portion 16k2, and the vertical portion to the third portion of the second portion 16k2. The distance y2 to the vertical portion of 16k3 (that is, the width of the third portion 16k3) is 1 mm or less, preferably 0.8 mm or less, and more preferably 0.5 mm or less.

Further, in the horizontal direction, the distance X from the vertical portion of the third portion 16k3 to the edge P2 of the upper surface 10a of the container bottom portion 10 adjacent via the groove portion 28 is 0.01 mm or more, and preferably 0.05 mm or more. Yes, more preferably 0.1 mm or more, still more preferably 0.4 mm or more. This is because the formed drop is easier to hold as the distance X increases, and sufficient hold is obtained when the distance X increases to a certain level. On the other hand, if the distance X is too large, the number of drop forming portions that can be installed in the limited space decreases, so it is preferably 4 mm or less.

The cell handling container 4 configured as described above can obtain the same operational effect as that of the above-described third embodiment, and further exhibits the following effect. That is, since the side surface 16k of the drop forming portion 16 is formed in a stepped shape, when the drop S formed on the upper surface 16a of the drop forming portion 16 spreads to the surroundings due to vibration or the like, the drop S has a second side surface 16k. It is possible to extend to the edge of the horizontal portion of the portion 16k2 or to the edge of the horizontal portion of the third portion 16k3 and stay there (see FIG. 8C). As a result, it is possible to prevent the drop S from collapsing. In this case, although the thickness of the drop S is reduced due to the spread of the drop S, it does not affect the culture or washing of the fertilized egg. In the present embodiment, the side surface 16k of the drop forming portion 16 is a stepped surface having three steps, but it may have two steps or four steps or more.

<Fifth Embodiment>
FIG. 9A is a plan view showing the cell handling container of the fifth embodiment, and FIG. 9B is a sectional view taken along line EE of FIG. 9A. The difference between the cell handling container 5 of the present embodiment and the first embodiment is the shape of the drop forming portion 30. Since other configurations are similar to those of the first embodiment, duplicated description will be omitted.

Specifically, the upper surface 10a of the container bottom 10 is provided with a plurality of (four in this embodiment) drop forming portions 30 for forming liquid drops such as a culture solution. These drop forming parts 30 have a cylindrical shape, are erected from the upper surface 10 a of the container bottom part 10 and are arranged at equal intervals around the cell containing area 12. The drop forming portion 30 is formed by a concave portion 31 that opens upward and a peripheral wall portion 32 that forms the concave portion 31.

The recess 31 has a rectangular internal space in a cross-sectional view, and the bottom surface 31a thereof is formed flat. The peripheral wall portion 32 has an inner wall surface 32a that forms an internal space of the recess 31, an outer wall surface 32b that is located outside the inner wall surface 32a, and an upper end surface 32c that is located above. The upper end surface 32c corresponds to the "upper end portion of the peripheral wall portion" recited in the claims and is a flat surface. The upper end surface 32c is, together with the concave portion 31, a liquid drop forming region. In this embodiment, the upper end surfaces 32c of the peripheral wall portions 32 of the four drop forming portions 30 are all flush with each other. On the other hand, since the outer wall surface 32b is arranged at a right angle to the upper end surface 32c, the angle α formed by the upper end surface 32c and the outer wall surface 32b at the edge P1 of the upper end surface 32c is 90°. The outer wall surface 32b extends in the vertical direction from the edge P1 of the upper end surface 32c to a range of 5 μm or more.

In the present embodiment, in the vertical direction, the distance from the upper end surface 32c of the peripheral wall portion 32 to the upper surface 10a of the container bottom portion 10 is h1, and the distance from the upper end surface 32c of the peripheral wall portion 32 to the bottom surface 31a (the deepest portion) of the recess 31. Where h2 is h2, the sum of h1 and h2 is less than 0.2 mm (that is, h1+h2<0.2 mm). The sum of h1 and h2 is preferably 0.1 mm or less, and more preferably the sum of h1 and h2 is 0.02 mm or less. In this embodiment, since the bottom surface 31a of the recess 31 is flush with the top surface 10a of the container bottom 10, h1=h2. Further, symbol f shown in FIG. 9B is the thickness of the peripheral wall portion in the horizontal direction (that is, the distance from the inner wall surface to the outer wall surface of the peripheral wall portion).

In the cell handling container 5 configured in this manner, the upper end surface 32c of the peripheral wall portion 32 and the recess 31 are liquid drop forming areas, and the outer wall surface 32b is in the range of 5 μm or more from the edge P1 of the upper end surface 32c in the vertical direction. Has been extended to. Therefore, similarly to the first embodiment, even if the upper end surface 32c is a hydrophilic surface, the drop S of the culture solution can be easily formed by the upper end surface 32c and the recess 31.

Further, since h1+h2<0.2 mm as described above, when the instrument such as a pipette is slid and moved, contact between the tip of the instrument and the inner wall surface 32a and the outer wall surface 32b of the peripheral wall portion 32 is suppressed. Therefore, it is possible to prevent damage due to contact as in the conventional case, and it is possible to improve workability.

Various modifications can be considered for the shapes of the peripheral wall and the recess. For example, in the modified example shown in FIG. 10A, the inner wall surface 32d of the peripheral wall portion 32 is an inclined surface that is inclined inward as it goes downward in a cross-sectional view. The bottom surface 31b of the recess 31 is a flat surface and is flush with the top surface 10a of the container bottom 10 (that is, h1=h2). The recess 31 has a cross section of an inverted truncated cone shape. At this time, at the edge P1 of the upper end surface 32c, the angle α formed by the upper end surface 32c and the outer wall surface 32b is 90°.

In the modification shown in FIG. 10B, in a cross-sectional view, the inner wall surface 32e of the peripheral wall portion 32 is an inclined surface that inclines inward as it goes downward, and the recess 31 has a V shape (that is, an inverted triangle that opens upward). Has become. The bottom surface 31c of the recess 31 is the apex of an inverted triangle and is flush with the top surface 10a of the container bottom 10 (that is, h1=h2). At this time, at the edge P1 of the upper end surface 32c, the angle α formed by the upper end surface 32c and the outer wall surface 32b is 90°.

In the modification shown in FIG. 11A, in cross-sectional view, the inner wall surface 32f of the peripheral wall portion 32 is a step surface having a vertical portion and an inclined portion in order from above. The vertical portion is a portion extending at a right angle to the upper end surface 32c, and the inclined portion is a portion inclined inward as it goes downward. At this time, the concave portion 31 has a shape in which an upper rectangular shape and a lower inverted triangle are combined in a sectional view. The bottom surface 31d of the recess 31 is the apex of an inverted triangle and is flush with the top surface 10a of the container bottom 10 (that is, h1=h2). At this time, at the edge P1 of the upper end surface 32c, the angle α formed by the upper end surface 32c and the outer wall surface 32b is 90°.

In the modification shown in FIG. 11B, the inner wall surface 32g of the peripheral wall portion 32 is formed so as to have a vertical portion and a curved portion in order from above in a cross-sectional view. The vertical portion is a portion that extends at a right angle to the upper end surface 32c, and the curved portion is a portion that is curved so as to be convex downward. Therefore, the bottom surface 31e of the recess 31 is a convex curved surface that is convex downward, and the deepest part thereof is located on the same plane as the top surface 10a of the container bottom 10 (that is, h1=h2). At this time, at the edge P1 of the upper end surface 32c, the angle α formed by the upper end surface 32c and the outer wall surface 32b is 90°.

In the modification shown in FIG. 12A, in a cross-sectional view, the inner wall surface 32e of the peripheral wall portion 32 is an inclined surface that inclines inward as it goes downward, and the recess 31 has a V shape (that is, an inverted triangle that opens upward). Has become. The bottom surface 31c of the recess 31 is the apex of an inverted triangle and is flush with the top surface 10a of the container bottom 10 (that is, h1=h2). On the other hand, at the upper end of the peripheral wall portion 32, the outer wall surface 32b and the inner wall surface 32e intersect with each other to form a ridge line portion 32h. The ridge line portion 32h corresponds to the "upper end portion of the peripheral wall portion" recited in the claims. Further, at the edge P1 of the ridgeline portion 32h, the angle α formed by the outer wall surface 32b and the inner wall surface 32e is an acute angle. A more acute angle α can form a stable drop due to the surface tension of the culture solution.

In the modification shown in FIG. 12B, in a cross-sectional view, the inner wall surface 32e of the peripheral wall portion 32 has an inclined surface that inclines inward as it goes downward, and the recess 31 has a V shape (an inverted triangle that opens upward). There is. The bottom surface 31c of the recess 31 is the apex of an inverted triangle and is flush with the top surface 10a of the container bottom 10 (that is, h1=h2). The outer wall surface 32i has an inclined portion that inclines outward as it goes downward and a vertical portion that extends in the vertical direction in order from the upper side. On the other hand, at the upper end of the peripheral wall portion 32, a ridge line portion 32h is formed by intersecting the inclined portion of the outer wall surface 32i and the inner wall surface 32e. The ridge line portion 32h corresponds to the "upper end portion of the peripheral wall portion" recited in the claims. Further, at the edge P1 of the ridgeline portion 32h, the angle α formed by the inclined portion of the outer wall surface 32i and the inner wall surface 32e is an acute angle.

In the modification shown in FIG. 13A, in a cross-sectional view, the inner wall surface 32e of the peripheral wall portion 32 has an inclined surface that inclines inward as it goes downward, and the recess 31 has a V shape (an inverted triangle that opens upward). There is. The bottom surface 31c of the recess 31 is the apex of an inverted triangle and is flush with the top surface 10a of the container bottom 10 (that is, h1=h2). The outer wall surface 32j is an inclined surface that inclines outward as it goes downward. On the other hand, at the upper end of the peripheral wall portion 32, the outer wall surface 32j and the inner wall surface 32e intersect with each other to form a ridge line portion 32h. The ridge line portion 32h corresponds to the "upper end portion of the peripheral wall portion" recited in the claims. At the edge P1 of the ridgeline portion 32h, the angle α formed by the outer wall surface 32j and the inner wall surface 32e is an acute angle.

In the modified example shown in FIG. 13B, in a cross-sectional view, the outer wall surface 32k and the inner wall surface 32l of the peripheral wall portion 32 have an inclined portion that inclines outward as it goes downward and a vertical portion that extends in the up-down direction in order from above. Are formed respectively. The upper end portion of the peripheral wall portion 32 is formed as a sharp portion by the inclined portion of the outer wall surface 32k and the inclined portion of the inner wall surface 32l intersecting in an inverted V shape. Therefore, the ridge line portion 32h is formed at the upper end of the peripheral wall portion 32. The ridge line portion 32h corresponds to the "upper end portion of the peripheral wall portion" recited in the claims. At the edge P1 of the ridgeline portion 32h, the angle α formed by the outer wall surface 32k and the inner wall surface 32l is an acute angle. Note that the symbol g shown in FIG. 13B is the distance of the pointed portion in the vertical direction (that is, the distance from the ridge line portion 32h to the boundary portion between the inclined portion and the vertical portion of the outer wall surface 32k (or the inner wall surface 32l)).

Although the bottom surface of the recess 31 is flush with the top surface 10a of the container bottom 10 in the present embodiment, the bottom surface 31a of the recess 31 is arranged higher than the top surface 10a of the container bottom 10 as shown in FIG. 14A. Alternatively, as shown in FIG. 14B, the bottom surface 31a of the recess 31 may be arranged lower than the top surface 10a of the container bottom 10.

<Sixth Embodiment>
FIG. 15A is a plan view showing the cell handling container of the sixth embodiment, and FIG. 15B is a sectional view taken along the line FF of FIG. 15A. The difference between the cell handling container 6 of the present embodiment and the first embodiment is the shape of the drop forming portion 40. Since other configurations are similar to those of the first embodiment, duplicated description will be omitted.

Specifically, the upper surface 10a of the container bottom 10 is provided with a plurality of (four in this embodiment) drop forming portions 40 for forming liquid drops such as a culture solution. Each of the drop forming portions 40 has a cylindrical shape, is formed so as to be embedded inside the container bottom portion 10, and is arranged at equal intervals around the cell containing area 12. The drop forming portion 40 is formed of a concave portion 41 that opens upward and a peripheral wall portion 42 that forms the concave portion 41. Further, a groove 43 surrounding the peripheral wall 42 is provided between the container bottom 10 and the peripheral wall 42 for each drop forming portion 40.

The recess 41 has a rectangular internal space in a cross-sectional view, and the bottom surface 41a thereof is formed flat. The peripheral wall portion 42 has an inner wall surface 42a forming an internal space of the recess 41, an outer wall surface 42b located outside the inner wall surface 42a, and an upper end surface 42c located above. Upper end surface 42c
Corresponds to the “upper end portion of the peripheral wall portion” described in the claims, is a flat surface, and is flush with the upper surface 10 a of the container bottom portion 10. The upper end surface 42c, together with the recess 41, serves as a liquid drop forming region. In the present embodiment, the upper end surfaces 42c of the peripheral wall portions 42 of the four drop forming portions 40 are all flush with each other and flush with the bottom surface 14a of the culture fluid storage portion 14 in the cell storage area 12. On the other hand, since the outer wall surface 42b is disposed at a right angle to the upper end surface 42c, the angle α formed by the upper end surface 42c and the outer wall surface 42b at the edge P1 of the upper end surface 42c is 90°. The outer wall surface 42b extends up to a range of 5 μm or more from the edge P1 of the upper end surface 42c in the vertical direction, and constitutes one side surface of the groove 43.

In the horizontal direction, the distance d from the edge P1 of the upper end surface 42c of the drop forming portion 40 to the edge P2 of the upper surface 10a of the container bottom portion 10 that is adjacent via the groove portion 43 is 0.01 mm or more, preferably 0.05 mm. It is above, more preferably 0.1 mm or more, still more preferably 0.4 mm or more. This is because the formed drop is easier to hold as the distance d increases, and sufficient hold is obtained when the distance d increases to a certain level. On the other hand, if the distance d is too large, the number of drop forming portions that can be installed in the limited space decreases, so that it is preferably 4 mm or less, for example. In this embodiment, the distance d is 0.5 mm.

In the present embodiment, the distance from the upper end surface 42c of the peripheral wall portion 42 to the upper surface 10a of the container bottom portion 10 is h3 (see FIG. 18) in the vertical direction, and the upper end surface 42c of the peripheral wall portion 42 to the bottom surface 41a of the recess 41 (the deepest position). When the distance to the portion is h4, the sum of h3 and h4 is 1 mm or less (that is, h3+h4≦1 mm). Preferably, the sum of h3 and h4 is 0.5 mm or less, more preferably the sum of h3 and h4 is 0.2 mm or less, and further preferably the sum of h3 and h4 is 0.1 mm or less, More preferably, the sum of h3 and h4 is 0.02 mm or less. In this embodiment, since the upper end surface 42c of the peripheral wall portion 42 and the upper surface 10a of the container bottom portion 10 are flush with each other, h3=0.

In the cell handling container 6 configured in this way, the upper end surface 42c of the peripheral wall portion 42 and the recess 41 are liquid drop forming regions, and the outer wall surface 42b is in the range of 5 μm or more from the edge P1 of the upper end surface 42c in the vertical direction. Has been extended to. Therefore, like the first embodiment described above, even if the upper end surface 42c is a hydrophilic surface, the drop S of the culture solution can be easily formed by the upper end surface 42c and the recess 41.

Further, as described above, since h3+h4≦1 mm (here, h3=0), when the instrument such as a pipette is slid and moved, the tip of the instrument and the inner wall surface 42a and the outer wall surface 42b of the peripheral wall portion 42 are Since such contact is suppressed, it is possible to prevent damage due to contact as in the conventional case, and it is possible to improve workability.

In the present embodiment, the bottom surface of the microwell 13 may be flush with the bottom surface 41 a of the recess 41 of the drop forming portion 40. By doing so, for example, when the cells C such as a fertilized egg are moved between the microwell 13 and the drop forming section 40, or when the cells C are arranged at both places, they are observed at the same focus position in the microscope observation. Therefore, the workability can be further improved.

Various modifications can be considered for the shapes of the peripheral wall and the recess. For example, in the modification shown in FIG. 16A, the inner wall surface 42d of the peripheral wall portion 42 is an inclined surface that inclines inward as it goes downward in a sectional view. The bottom surface 41b of the recess 41 is a flat surface. The cross section of the recess 41 has an inverted truncated cone shape that opens upward. At this time, at the edge P1 of the upper end surface 42c, the angle α formed by the upper end surface 42c and the outer wall surface 42b is 90°.

In the modification shown in FIG. 16B, in a cross-sectional view, the inner wall surface 42e of the peripheral wall portion 42 is an inclined surface that inclines inward as it goes downward, and the recess 41 has a V shape (an inverted triangle that opens upward). Has become. Therefore, the bottom surface 41c of the recess 41 is the apex of an inverted triangle. At this time, at the edge P1 of the upper end surface 42c, the angle α formed by the upper end surface 42c and the outer wall surface 42b is 90°.

In the modification shown in FIG. 17A, the inner wall surface 42f of the peripheral wall portion 42 is a stepped surface having a vertical portion and an inclined portion in order from above in a sectional view. The vertical portion is a portion that extends at a right angle to the upper end surface 42c, and the inclined portion is a portion that is inclined inward as it goes downward. At this time, the recess 41 has a shape in which the upper rectangular shape and the lower inverted triangle are combined in a cross-sectional view. Therefore, the bottom surface 41d of the recess 41 is the apex of an inverted triangle. At this time, at the edge P1 of the upper end surface 42c, the angle α formed by the upper end surface 42c and the outer wall surface 42b is 90°.

In the modification shown in FIG. 17B, the inner wall surface 42g of the peripheral wall portion 42 is a curved surface that is convex downward in a cross-sectional view. Therefore, the bottom surface 41e of the concave portion 31 is a convex curved surface that is convex downward. At this time, at the edge P1 of the upper end surface 42c, the angle α formed by the upper end surface 42c and the outer wall surface 42b is 90°.

In the present embodiment, it has been described that the upper end surface 42c of the peripheral wall portion 42 is flush with the upper surface 10a of the container bottom portion 10. However, as shown in FIG. 18A, the upper end surface 42c of the peripheral wall portion 42 is located above the upper surface of the container bottom portion 10. It may be arranged higher than 10a, or, as shown in FIG. 18B, the upper end surface 42c of the peripheral wall portion 42 may be arranged lower than the upper surface 10a of the container bottom portion 10.

<Seventh Embodiment>
FIG. 19A is a partial plan view showing the cell handling container of the seventh embodiment, and FIGS. 19B and 19C are sectional views taken along the line GG of FIG. 19A. The difference between the cell handling container 7 of the present embodiment and the sixth embodiment is that a further surrounding wall portion 44 surrounding the drop forming portion 40 is further provided and a groove portion 45 is provided between the container bottom portion 10 and the surrounding wall portion 44. Is. Other configurations are the same as those in the sixth embodiment, and thus redundant description will be omitted.

Specifically, an annular surrounding wall portion 44 is provided on the outer periphery of the peripheral wall portion 42 of the drop forming portion 40 so as to surround the peripheral wall portion 42. The surrounding wall portion 44 is arranged at a predetermined distance from the outer wall surface 42b of the peripheral wall portion 42. The upper end surface 44a of the surrounding wall portion 44 is flush with the upper end surface 42c of the peripheral wall portion 42 and the upper surface 10a of the container bottom portion 10. In the horizontal direction, the distance c from the edge P1 of the upper end surface 42c of the peripheral wall portion 42 to the outer edge P3 of the upper end surface 44a of the surrounding wall portion 44 is 1 mm or less, preferably 0.8 mm or less, and 0.5 mm or less. Is more preferable.

Further, a groove 45 surrounding the surrounding wall 44 is provided between the surrounding wall 44 and the container bottom 10. The groove portion 45 is formed in a U-shape whose cross section opens upward. One of the side surfaces of the groove portion 45 is formed by the side surface of the surrounding wall portion 44.

Further, in the horizontal direction, the distance d1 from the edge P1 of the upper end surface 42c of the peripheral wall portion 42 to the closest edge P4 of the upper end surface 44a of the surrounding wall portion 44, and the groove portion 45 from the outer edge P3 of the upper end surface 44a of the surrounding wall portion 44. The distance d2 to the edge P2 of the upper surface 10a of the container bottom 10 adjacent to each other is 0.01 mm or more, preferably 0.05 mm or more, more preferably 0.1 mm or more, and further preferably It is 0.4 mm or more. This is because when the distances d1 and d2 described above are large to some extent, the drop is easily held. On the other hand, if the distances d1 and d2 are too large, the number of drop forming portions that can be installed in the limited space decreases, so it is preferable that each is 4 mm or less.

The cell handling container 7 configured as described above can obtain the same operational effect as that of the above-described sixth embodiment. Further, since the surrounding wall portion 44 surrounding the drop forming portion 40 is provided and the groove portion 45 is provided between the surrounding wall portion 44 and the container bottom portion 10, the drop S formed by the recess 41 and the upper end surface 42c of the peripheral wall portion 42 is provisionally formed. When S spreads around due to vibration or the like, the drop S can spread to the surrounding wall portion 44 and stay on the outer edge P3 of the surrounding wall portion 44 (see FIG. 19C). As a result, it is possible to prevent the drop S from collapsing. In this case, although the thickness of the drop S is reduced due to the spread of the drop S, it does not affect the culture or washing of the cell C such as a fertilized egg.

<Eighth Embodiment>
20A is a partial plan view showing the cell handling container of the eighth embodiment, and FIGS. 20B and 20C are cross-sectional views taken along the line HH of FIG. 20A. The difference between the cell handling container 8 of the present embodiment and the sixth embodiment is that the outer wall surface 42h of the peripheral wall portion 42 is formed by two or more stepped surfaces. Other configurations are the same as those in the sixth embodiment, and thus redundant description will be omitted.

Specifically, the outer wall surface 42h of the peripheral wall portion 42 extends downward along the entire circumference of the upper end surface 42c, and is formed in a step shape (here, three steps) so as to expand from the inner side to the outer side. There is. The outer wall surface 42h includes, in order from above, a first portion 42h1 arranged at right angles to the upper end surface 42c, a second portion 42h2 having a horizontal portion and a vertical portion connected to the first portion 42h1, and a second portion 42h2. The third portion 42h3 is connected to the portion 42h2 and has a horizontal portion and a vertical portion.

In the vertical direction, a distance (that is, the height of the first portion 42h1) t1 from the edge P1 of the upper end surface 42c of the peripheral wall portion 42 to the horizontal portion of the second portion 42h2 of the outer wall surface 42h, from the horizontal portion of the second portion 42h2. The distance t2 to the horizontal portion of the third portion 42h3 (that is, the height of the second portion 42h2) and the distance from the horizontal portion of the third portion 42h3 to the bottom surface of the groove portion 46 (described later) (that is, the third portion). The height t3 of 42h3 is 5 μm or more. The heights t1, t2 and t3 are preferably 10 μm or more, more preferably 50 μm or more, further preferably 100 μm or more, and further preferably 250 μm or more.

These lower limit values are because the higher the above-mentioned heights t1, t2, and t3, the easier it is for the formed drops to be held, and when the heights reach a certain level, sufficient holding is obtained. For example, when the heights t1, t2, and t3 are each 50 μm or more, a thick drop can be created even if the liquid amount is increased, and when the heights t1, t2, and t3 are 100 μm or more, a thick drop is generated even if the liquid amount is increased. It is possible to stably produce, and when it is 250 μm or more, a thick drop can be produced more stably even if the liquid amount is increased. The upper limits of the heights t1, t2, t3 are not particularly limited, but the higher the height, the more difficult it is to form from the viewpoint of manufacturing.

At this time, since the first portion 42h1 of the outer wall surface 42h is arranged at a right angle to the upper end surface 42c, the angle formed by the upper end surface 42c and the first portion 42h1 of the outer wall surface 42h at the edge P1 of the upper end surface 42c. α is 90°.

Further, an annular groove portion 46 surrounding the peripheral wall portion 42 is provided between the peripheral wall portion 42 and the container bottom portion 10. In cross-sectional view, the groove portion 46 is opened upward, one of the side surfaces of the groove portion 46 is formed stepwise by the outer wall surface 42h of the peripheral wall portion 42, and the other is a vertical surface.

In the horizontal direction, the distance (that is, the width of the second portion 42h2) y1 from the edge P1 of the upper end surface 42c of the peripheral wall portion 42 to the vertical portion of the second portion 42h2, and the vertical portion to the third portion of the second portion 42h2. The distance y2 to the vertical portion of 42h3 (that is, the width of the third portion 42h3) y2 is 1 mm or less, preferably 0.8 mm or less, and more preferably 0.5 mm or less.

Further, in the horizontal direction, the distance X from the vertical portion of the third portion 42h3 to the edge P2 of the upper surface 10a of the container bottom portion 10 adjacent to the groove portion 46 is 0.01 mm or more, preferably 0.05 mm or more. Yes, more preferably 0.1 mm or more, still more preferably 0.4 mm or more. This is because the formed drop is easier to hold as the distance X increases, and sufficient hold is obtained when the distance X increases to a certain level. On the other hand, if the distance X is too large, the number of drop forming portions that can be installed in the limited space decreases, so it is preferably 4 mm or less.

The cell handling container 8 configured as described above can obtain the same operational effect as that of the above-described sixth embodiment, and further exhibits the following effect. That is, since the outer wall surface 42h of the peripheral wall portion 42 is formed in a stepped shape, when the drop S formed by the recess 41 and the upper end surface 42c of the peripheral wall portion 42 spreads to the surroundings due to vibration or the like, the drop S is formed on the outer wall surface. It is possible to spread to the edge of the horizontal portion of the second portion 42h2 of 42h or to the edge of the horizontal portion of the third portion 42h3 and stay there (see FIG. 20C). As a result, it is possible to prevent the drop S from collapsing. In this case, although the thickness of the drop S is reduced due to the spread of the drop S, it does not affect the culture or washing of the cell C such as a fertilized egg. In addition, in the present embodiment, the outer wall surface 42h of the peripheral wall portion 42 has three stepped surfaces, but may have two steps or four steps or more.

<Ninth Embodiment>
21A is a plan view showing a cell handling container (a state in which a lid is removed) of the ninth embodiment, FIG. 21B is a sectional view taken along line I-I of FIG. 21A, and FIG. 21C is J of FIG. 21A. 21D is a sectional view taken along line JJ of FIG. 21A, and FIG. 21E is a sectional view taken along line LL of FIG. 21A. The difference between the cell handling container 9 of the present embodiment and the above-described embodiment is the structure in which a plurality of types (here, four types) of drop forming portions are combined. Other configurations are the same as those in the above-described embodiment, and thus redundant description will be omitted.

Specifically, on the upper surface 10a of the container bottom 10, drop forming portions having different structures are provided around the cell containing area 12 one by one. A structure having the drop forming portion 16 and the groove portion 17 according to the second embodiment at the place M1 shown in FIG. 21A (see FIG. 21B), and the drop forming portion 16 and the surrounding wall portion 26 and the groove portion 27 according to the third embodiment at the place M2. The structure having (see FIG. 21C) and the structure having the drop forming portion 30 of the fifth embodiment (see FIG. 21D) are arranged at the place M3.

On the other hand, in the structure provided in the place M4, in addition to the drop forming portion 30 of the fifth embodiment, the surrounding wall portion 33 surrounding the drop forming portion 30 is provided. The upper end surface 33a of the surrounding wall portion 33 is flush with the upper end surface 32c of the peripheral wall portion 32 (see FIG. 21E).

According to the cell handling container 9 configured as described above, it is possible to easily form a drop in each drop forming portion even on a hydrophilic surface, and in addition, a plurality of types of drops can be placed in one cell handling container 9. By providing the forming section, by properly using these drop forming sections as necessary, one cell handling container 9 can meet various needs, and the versatility of the cell handling container 9 can be enhanced.

<Tenth Embodiment>
22A is a plan view showing the cell handling container of the tenth embodiment, and FIG. 22B is a sectional view taken along line MM of FIG. 22A. The difference between the cell handling container 53 of the present embodiment and the second embodiment is that the cell forming area 12 is not provided, and instead, the drop forming portion 54 is provided at the center of the container bottom 10. Since other configurations are the same as those in the second embodiment, duplicate description will be omitted.

Specifically, in the center of the container bottom 10, there is provided a drop forming portion 54 having an upper surface 54a serving as a drop forming area and a side surface 54b extending downward along the entire circumference of the upper surface 54a. The drop forming portion 54 is formed in a cylindrical shape similarly to the four drop forming portions 16 arranged around the drop forming portion 54, but is formed wider than the drop forming portion 16. The upper surface 54a of the drop forming portion 54 is a substantially flat surface and is flush with the upper surface 16a of the drop forming portion 16 and the upper surface 10a of the container bottom 10.

In the center of the drop forming portion 54, a plurality of microwells 55 are formed that are recessed inward from the upper surface 54a. Each of these microwells 55 has a substantially columnar shape so as to be suitable for accommodating and culturing cells C such as fertilized eggs, and its bottom surface is a conical surface inclined from the outside toward the center. Note that the microwell 55 may have a U-shaped cross section with an upward opening, a U-shaped cross section having an arc portion, or other shapes, as in the microwell 13 of the first embodiment. The side surface 54b of the drop forming portion 54 is arranged at a right angle to the upper surface 54a.

Further, an annular groove portion 56 surrounding the drop forming portion 54 is provided between the drop forming portion 54 and the container bottom portion 10. The groove portion 56 is formed in a U-shape whose cross section opens upward. The side surface 54b of the drop forming portion 54 constitutes one of the side surfaces of the groove portion 56. The height of the upper surface 54a of the drop forming portion 54 with respect to the bottom surface of the groove portion 56 is the same as the height t of the upper surface 16a of the drop forming portion 16 with respect to the bottom surface of the groove portion 17. Further, the distance from the edge of the upper surface 54a of the drop forming portion 54 to the edge of the upper surface 10a of the container bottom portion 10 that is adjacent via the groove portion 56 is adjacent from the edge P1 of the upper surface 16a of the drop forming portion 16 via the groove portion 17. It is the same as the distance d to the edge P2 of the upper surface 10a of the container bottom 10.

The cell handling container 53 thus configured can obtain the same operational effect as that of the above-described second embodiment, and further exhibits the following effect. That is, since the drop forming portion 54 is provided, it is possible to increase the number of drop forming locations. Further, since the upper surface 54a of the drop forming portion 54, the upper surface 16a of the drop forming portion 16 and the upper surface 10a of the container bottom portion 10 are flush with each other and there is no barrier, a device such as a pipette can be slid and moved. The workability can be further improved.

<Eleventh Embodiment>
FIG. 23A is a plan view showing the cell handling container of the eleventh embodiment, and FIG. 23B is a sectional view taken along the line NN of FIG. 23A. The difference between the cell handling container 57 of the present embodiment and the tenth embodiment is that a surrounding wall portion 58 surrounding the drop forming portion 54 is further provided, and a groove portion 59 is provided between the container bottom portion 10 and the surrounding wall portion 58. Is. Since other configurations are the same as those of the tenth embodiment, duplicate description will be omitted.

Specifically, an annular surrounding wall portion 58 is provided on the outer periphery of the drop forming portion 54 so as to surround the drop forming portion 54. The surrounding wall portion 58 is arranged at a predetermined distance from the side surface 54b of the drop forming portion 54. The upper end surface 58a of the surrounding wall portion 58 is flush with the upper surface 54a of the drop forming portion 54 and the upper surface 10a of the container bottom portion 10. The distance from the edge of the upper surface 54a of the drop forming portion 54 to the outer edge of the upper end surface 58a of the surrounding wall portion 58, the distance from the edge of the upper surface 54a to the nearest edge of the upper end surface 58a of the surrounding wall portion 58, the surrounding wall portion 58 The distance from the outer edge of the upper end surface 58a to the edge of the upper surface 10a of the container bottom portion 10 that is adjacent via the groove 59 is the same as in the third embodiment described above, so description thereof will be omitted.

The cell handling container 57 thus configured can obtain the same operational effect as that of the above-described tenth embodiment.

<Twelfth Embodiment>
FIG. 24 is a perspective view showing the cell handling container of the twelfth embodiment (with the lid removed). The cell handling container 60 of the present embodiment includes a bottomed cylindrical container body 1a that opens upward, and a lid (not shown) that is detachably installed on the upper portion of the container body 1a. The side wall portion 11 of the container body 1a is located above the thick wall portion 11a and is formed to be relatively thick by extending from the upper surface 10a of the container bottom portion 10 toward the opening. It is composed of a thin portion 11b formed thinner than the portion 11a. A step portion 11c is formed at a substantially intermediate position of the side wall portion 11 due to a change in thickness between the thick portion 11a and the thin portion 11b. The step portion 11c is a portion that comes into contact with the edge portion of the lid when the lid is put on the container body 1a.

A cell storage area 12 is provided at the center of the container bottom 10. Further, the container bottom portion 10 has a structure including the drop forming portion 16, the surrounding wall portion 26 and the groove portion 27 as in the above-described third embodiment (hereinafter, in order to avoid complexity of the description, the structure is referred to as “with surrounding wall portion and groove portion”). 6) are provided. These surrounding wall portions and grooved structures are regularly arranged with three pieces on each side (three pieces on the upper side and three pieces on the lower side with respect to the cell containing area 12 in FIG. 24) centering on the cell containing area 12.

FIG. 25 is a plan view showing the container body of the cell handling container. As shown in FIG. 25, when the drop forming portion 16 approaches the side wall portion 11, it becomes difficult to operate a device such as a pipette. Therefore, the shortest distance L1 from the surrounding wall portion and the groove portion 27 of the grooved structure to the side wall portion 11 is to some extent. You need to leave. From the viewpoint of manufacturing, the shortest distance L2 between adjacent wall portions and structures with groove portions (that is, the shortest distance between adjacent groove portions 27), and the groove portion 27 of the wall portions and grooved structure from the cell accommodation area 12 The shortest distance L3 to the outer wall is preferably 0.5 mm or more, and more preferably 1.0 mm or more. With this configuration, when the cell C such as the fertilized egg is washed with a device such as a pipette, these devices can be arranged at positions where they can be easily operated. Further, in the container bottom portion 10, a space excluding the surrounding wall portion and the grooved structure, the cell containing area 12, and the outer wall well 61 becomes a plurality of free spaces W. Since the operator can selectively use these free spaces W, the versatility of the cell handling container 60 can be enhanced.

Further, the container bottom portion 10 is provided with a curved wall 62 that stands up from the upper surface 10a and is curved in a C shape on the side wall portion 11 side. Both left and right ends of the curved wall 62 extend to the side wall portion 11 so as to contact the inside of the side wall portion 11. The space surrounded by the side wall portion 11 and the curved wall 62 constitutes the outer wall well 61. The curved wall 62 has an arc shape, and is configured by a part such as a circle or an ellipse. In the present embodiment, the curved wall 62 is a part of an ellipse having a major axis of 12 mm and a minor axis of 6 mm.

FIG. 26 is a sectional view taken along the line PP of FIG. As shown in FIG. 26, the curved wall 62 has a uniform thickness and height, and its cross section has a trapezoidal shape whose width decreases from the lower side to the upper side. As the dimensions, for example, when the container body 1a has an inner diameter of 35 mm, the height of the curved wall 62 is 0.5 mm, the lower bottom is 0.5 mm, and the angle α″ of the side surface of the trapezoid with respect to the vertical direction (that is, the vertical direction) is It is 3°.

In addition, a recess 63 is provided on the bottom surface 10b of the container bottom 10. As shown in FIG. 27A, the recess 63 is recessed inside the container bottom 10 and has a substantially U-shape in a bottom view. In the present embodiment, the recess 63 is, for example, a part of an ellipse having a major axis of 6 mm and a minor axis of 3 mm. As shown in FIG. 27B, the recess 63 is recessed by 3 mm inside the container bottom 10 and extends upward from the bottom surface 10b to a height of 5 mm.

In the cell handling container 60 configured as described above, the outer wall well 61 is provided in the container bottom portion 10. Therefore, a liquid such as a culture solution is put into the outer wall well 61 to wash the tip of an instrument such as a pipette (that is, (Co-washing), or cells C such as fertilized eggs can be placed with a pipette or the like for washing or culturing. Further, even if the height of the curved wall 62 forming the outer wall well 61 is low (for example, 0.5 mm described above), the liquid volume (for example, 10 μL or more and 30 μL or less) necessary for co-washing or washing or culturing of the cells C can be maintained. Not only can it be held in the outer wall well 61, but also a liquid height (for example, 1.0 mm or more) that can be easily operated when operating a device such as a pipette can be realized. Furthermore, as shown in FIG. 25, since the outer wall well 61 is arranged on one side of the container bottom portion 10, the empty space including the free space W can be utilized to the maximum extent, and instruments such as pipettes can be operated sequentially from the top. Therefore, the work can be efficiently performed.

Further, since the recessed portion 63 is provided on the bottom surface 10b of the container bottom portion 10, it can be used as a marking for visually identifying the recessed portion 63. Therefore, for example, at the time of microscopic observation, the direction and position of the cell handling container 60 can be easily grasped by using the recess 63 as a mark. Further, since the operator can grasp the direction and the position of the cell handling container 60 by touching the recess 63 with his/her finger, it is possible to perform the work by touch without touching the eyes, thereby improving the work efficiency. it can. Furthermore, when a protrusion or the like that can be engaged with the recess 63 is provided on the mounting surface of the microscope, the position of the cell handling container 60 can be easily fixed by engaging the recess 63 and the protrusion. , It is possible to reduce the risk that the observation position shifts.

<Thirteenth Embodiment>
28A is a plan view showing the cell handling container of the thirteenth embodiment, FIG. 28B is a sectional view taken along line RR of FIG. 28A, and FIG. 29A is an enlarged sectional view of a drop forming portion. The difference between the cell handling container 65 of the present embodiment and the second embodiment is that the upper surface 64a and the side surface 64c of the drop forming portion 64 are not directly connected but are connected via a curved surface portion 64b having a predetermined curvature radius R. Is to be done. Since other configurations are the same as those in the second embodiment, duplicate description will be omitted.

Specifically, the drop forming portion 64 has a flat upper surface 64a serving as a liquid drop forming region, and a side surface 64c connected to the upper surface 64a via a curved surface portion 64b and extending downward. In FIG. 29A, P6 indicates a boundary point between the upper surface 64a and the curved surface portion 64b (that is, the beginning of the curved surface portion 64b), and P7 indicates a boundary point between the side surface 64c and the curved surface portion 64b (that is, the end portion of the curved surface portion 64b). .. The drop S formed on the upper surface 64a is arranged within the range surrounded by the boundary point P6.

Further, in FIG. 29A, P5 is an intersection of a straight line along the upper surface 64a and a straight line along the side surface 64c. The angle α'is an angle formed by a straight line along the upper surface 64a and a straight line along the side surface 64c. In this embodiment, since the side surface 64c is perpendicular to the upper surface 64a, α'=90°. On the other hand, as shown in FIG. 29B, when the side surface 64c is inclined so as to expand outward as it goes downward, the angle α'is an obtuse angle.

The diameter D′, which is the minimum width of the upper surface 64a, is preferably 0.5 mm or more and 10 mm or less, more preferably 3 mm or more and 6 mm or less, as in the above embodiment. The height t′ of the upper surface 64a of the drop forming portion 64 with respect to the bottom surface of the groove portion 17 is 5 μm or more as in the second embodiment, preferably 10 μm or more, more preferably 50 μm or more, further preferably 100 μm or more, and 250 μm or more. Is most preferred.

Further, in the horizontal direction, the distance d′ from the intersection P5 of the straight line along the upper surface 64a and the straight line along the side surface 64c to the edge P2 of the upper surface 10a of the container bottom portion 10 adjacent via the groove portion 17 is the same as that of the second embodiment. Similarly, it is 0.01 mm or more, preferably 0.05 mm or more, more preferably 0.1 mm or more, and further preferably 0.4 mm or more. Further, if the distance d′ is too large, the number of drop forming portions 64 that can be installed in the limited space decreases, so the distance d′ is preferably 4 mm or less.

In the present embodiment, the radius of curvature R of the curved surface portion 64b is less than 250 μm. With this configuration, the drop formed on the upper surface 64a can be easily held. The radius of curvature R of the curved surface portion 64b is preferably 100 μm or less, more preferably 70 μm or less, further preferably 50 μm or less, and most preferably 20 μm or less.

According to the cell handling container 65 configured as described above, the same operational effect as that of the second embodiment can be obtained. The drop forming portion 64 having the curved surface portion 64b is also applied to the above-described first, third, fourth, tenth, eleventh, and twelfth embodiments, and detailed description thereof will be omitted. Note that, in the present embodiment, the example in which the upper surface 64a of the drop forming portion 64 is the liquid drop forming area has been described, but the upper surface 64a and a part of the curved surface portion 64b may be the liquid drop forming area. good. In this case, the drop S is formed so as to extend beyond the boundary point P6 to a part of the curved surface portion 64b.

<Fourteenth Embodiment>
FIG. 37A is a plan view showing the cell handling container of the fourteenth embodiment, and FIG. 37B is a sectional view taken along the line BB of FIG. 37A. The difference between the cell handling container 66 of the present embodiment and the second embodiment is that the side surface 16'b of the drop forming portion 16' is intermittently provided along the circumferential direction of the upper surface 16'a. Since other configurations are the same as those in the second embodiment, duplicate description will be omitted.

Specifically, the drop forming portion 16 ′ is arranged intermittently along the circumferential direction of the flat upper surface 16 ′ a that is a liquid drop forming region, and extends downward. And a side surface 16'b. Due to the intermittent arrangement of the side surfaces 16'b, a plurality of recesses 17' are formed in the circumferential direction of the upper surface 16'a. As shown in FIG. 37A, these recesses 17' each have a substantially fan shape in a plan view, and are arranged at equal intervals along the circumferential direction of the upper surface 16'a. The side surface 16'b provided intermittently extends downward from the upper surface 16'a to a range of 5 μm or more, and constitutes a part of the side surface of the recess 17'.

In the cell handling container 66 configured as described above, the side surface 16'b of the drop forming portion 16' is intermittently provided along the circumferential direction of the upper surface 16'a, but the side surface 16'b is the upper surface 16'. Since it extends downward from 5'or more to the range of 5'm or more, even if the upper surface 16'a is a hydrophilic surface, it is easy to drop the culture solution S on the upper surface 16'a as in the second embodiment. Can be formed. Note that, since other operational effects are the same as those of the second embodiment, redundant description will be omitted.

Hereinafter, the present invention will be described with reference to examples, but the present invention is not limited to the scope of the examples.

<Example 1>
In this example, using 9 kinds of samples (made of polystyrene) shown in Table 1, 7.5 μL of water was dropped after the hydrophilic treatment by plasma (water contact angle was 10° or less), and a drop was performed. It was verified whether it could be formed.

In Table 1 and all the following tables, t is the height of the upper surface of the drop forming portion, D is the diameter of the upper surface of the drop forming portion, d is the bottom of the container that is adjacent to the edge of the upper surface of the drop forming portion through the groove. The distance to the top edge is shown. Further, h1 represents the distance from the upper end of the peripheral wall to the upper surface of the container bottom, and f represents the thickness of the peripheral wall in the horizontal direction (that is, the distance from the inner wall surface to the outer wall surface of the peripheral wall).

As a result, any of the samples listed in Table 1 was able to form drops. 32 shows the sample No. 3 and No. 4 is a photograph showing the result of No. 4.

<Comparative Example 1>
As a comparative example, in a flat plate made of the same material (polystyrene), 7.5 μL of water was dropped after hydrophilic treatment by plasma (water contact angle of 10° or less), and as a result, the drop was crushed and a drop was created. I couldn't.

<Example 2>
In this embodiment, No. 1 in the first embodiment is used. Using 8 kinds of samples (polystyrene material) excluding 5, it was checked whether or not each liquid volume of water could be dropped after the hydrophilic treatment by plasma (water contact angle is 10° or less). Verified. Table 2-1 shows the results of the samples (No. 1, 2, 6, 7) corresponding to the drop forming portions of the first embodiment, and Table 2-2 shows the samples (No. 1 corresponding to the drop forming portions of the second embodiment. . 3, 4, 8, 9) are shown respectively.

As shown in Table 2-1, when the height t of the upper surface of the drop forming portion was 10 μm or more, drops (liquid volume 7.5 μL) could be formed in all the samples. Further, when the height t of the upper surface of the drop forming portion was 50 μm or more (Sample Nos. 2, 6 and 7), drops having a liquid volume of 10 μL or more and 20 μL or less could be formed. Furthermore, when the height t is 100 μm or more (No. 6, 7), a 10 μL drop can be formed, and when the height t is 250 μm or more (No. 7), a 15 μL drop can be formed by one operation. did it.

As shown in Table 2-2, when the height t of the upper surface of the drop forming portion was 10 μm or more, drops (liquid volume 7.5 μL) could be formed in all the samples. Further, when the height t of the upper surface of the drop forming portion was 50 μm or more (Sample Nos. 9, 3 and 4), drops having a liquid volume of 10 μL or more and 20 μL or less could be formed. Further, when the height t is 100 μm or more (No. 3, 4), 10 μL drop can be formed, and when the height t is 250 μm or more (No. 4), 15 μL drop can be formed by one operation. did it.

<Example 3>
In this embodiment, as shown in FIG. 9B, FIG. 14A, and FIG. 14B described above, the drop forming portion has a concave portion and a peripheral wall portion forming the concave portion, and the bottom surface of the concave portion is the same as or higher than the upper surface of the container bottom portion. Alternatively, the samples shown in Table 3 were prepared by using the samples arranged in a low position. For each sample, the operability of the pipette was verified according to the procedure of raising the pipette along the outer wall surface of the peripheral wall portion, sliding the pipette inside the peripheral wall portion, and further descending along the inner wall surface to the bottom surface of the recess.

As a result of the verification, as shown in Table 3, the sum of the distance h1 from the upper end of the peripheral wall to the upper surface of the container bottom and the distance h2 from the upper end of the peripheral wall to the bottom of the recess is less than 0.2 mm ( That is, if h1+h2<0.2 mm), the tip of the pipette hardly hit the peripheral wall (no contact). It was confirmed that the operability of the pipette was improved.

<Example 4>
In this example, the calculated value and the actually measured value regarding the thickness h′ of the formed drop were compared. The calculated value of the drop thickness h′ is calculated based on h′=3V/(2πr′ ² ). Here, r′ is the radius of the upper surface of the drop forming portion, that is, r′=D/2 (D is the diameter of the upper surface of the drop forming portion), h′≦r′, V is the culture solution volume [μL], π indicates the pi respectively. Since h′≦r′, the upper limit of V that can form a drop is V≦2πr′ ³
/3. On the other hand, the measured value of the thickness h′ of the drop was measured by using the samples (made of polystyrene) shown in Table 4 and dropping each liquid amount of water after the hydrophilic treatment by plasma (water contact angle was 10° or less). Then, the formed drop is actually measured.

As shown in Table 4, for the thickness h'of the formed drop, there was almost no difference between the calculated value based on the above formula and the actually measured value. Therefore, when the upper surface of the drop forming portion is circular, it was confirmed that the thickness h'of the formed drop satisfies the above formula.

<Example 5>
In this example, when the upper surface of the drop forming portion was circular, the influence of the difference in the diameter D on the drop formation was evaluated. Specifically, each sample formed of polystyrene as shown in Table 5 was evaluated by dropping each amount of water after the hydrophilic treatment with plasma (water contact angle of 10° or less). ..

As shown in Table 5, the larger the diameter D of the upper surface of the drop forming portion, the larger the capacity of the drop that can be formed, and the larger the capacity, the larger the thickness of the drop, so that the drop can be formed by one operation. It was confirmed that the range was widened. On the other hand, in the comparison when the same amount of liquid was added, the smaller the diameter D, the thicker the drop.

<Comparative example 2>
When culturing or washing cells such as fertilized eggs, the volume of the culture solution is used in the range of 15 μL or more and 30 μL or less, and the thickness of the formed drop is 1 mm or more from the viewpoint of ensuring good operability of instruments such as pipettes. Is preferable, and 1.5 mm or more is more preferable. However, on a flat surface having a water contact angle θ of less than 80° in the range of the culture solution volume of 15 μL or more and 30 μL or less, the thickness of the drop is 1.5 mm or less, which makes it difficult to operate the device. On a flat surface having a water contact angle θ of 60° or less, the thickness of the drop is 1.0 mm or less, which makes it very difficult to operate the device. In this comparative example, the drop thickness due to the water contact angle when the drop was formed on a flat surface (material: polystyrene) was examined (see Table 6).

<Example 6>
In this example, in the drop forming portion of the above-described second embodiment (see FIG. 3B), the drop thickness according to the water contact angle at the liquid amounts of 15 μL and 30 μL was examined as in Comparative Example 2 (see Table 7). In the evaluation sample having a liquid volume of 15 μL, t=250 μm, D=4.0 mm, d=0.5 mm, and in the evaluation sample having a liquid volume of 30 μL, t=250 μm, D=5.0 mm, d=0.5 mm. there were.

As can be seen from Table 6 and Table 7, in Example 6, for example, in the range of 15 μL or more and 30 μL or less, the water contact angle is less than 80°, further 60° or less, and even 10° or less, the drop thickness is 1. Since it was 5 mm or more, the result was that instruments such as pipettes were easy to operate.

<Example 7>
In the present embodiment, for the sample of the drop forming portion of the second embodiment shown in Table 8 (material: polystyrene, after hydrophilic treatment by plasma (water contact angle is 10° or less)), from the edge of the upper surface of the drop forming portion The influence of the difference in the size of the distance d to the edge of the upper surface of the container bottom adjacent to the groove via the groove was evaluated.

From the results shown in Table 8, it was confirmed that drops can be formed in the range of d of 0.4 mm or more and 1.0 mm or less, and the drop formation is not affected by the difference in d size.

<Example 8>
In this example, a sample (material: polystyrene, after hydrophilic treatment by plasma (water contact angle is 10° or less), liquid amount: 30 μL) of the drop forming portion (see FIGS. 7A to 7C) of the third embodiment is used. Then, vibration was applied after forming the drops, and changes in the drops before and after the vibration were evaluated. The dimensions of the sample are shown in Table 9.

FIG. 30A is a photograph showing the results of Example 8. In FIG. 30A, after the vibration is applied on the left side, the photographs before the vibration are applied on the right side, respectively. From the photograph, it was confirmed that the drop did not collapse even when it was vibrated and stayed on the surrounding wall.

<Example 9>
In this example, a sample (material: polystyrene, after hydrophilic treatment by plasma (water contact angle is 10° or less), liquid amount: 15 μL) of the drop forming portion (see FIGS. 8A to 8C) of the fourth embodiment is used. It was verified whether or not the drop could be formed. The dimensions of the sample are shown in Table 10.

As a result, it was confirmed that drops could be formed.

<Example 10>
In this example, the conditions shown in Table 11 were used using the sample (material: polystyrene, after hydrophilic treatment by plasma (water contact angle was 10° or less)) of the drop forming portion (see FIG. 9B) of the fifth embodiment. It was verified whether or not a drop can be formed, and when the drop can be formed, the influence on the drop due to the difference in size of h1=h2 was evaluated.

In Table 11, h1 indicates the distance from the upper end surface of the peripheral wall portion to the upper surface of the container bottom portion, h2 indicates the distance from the upper end surface of the peripheral wall portion to the bottom surface of the recess, and f indicates the thickness of the peripheral wall portion in the horizontal direction. From the results in Table 10, when h1 was 50 μm or more, drops (liquid volume 40 μL) could be formed in all the samples. Further, when h1 was 250 μm or more, drops having a liquid volume of 50 μL or more and 80 μL or less could be formed.

<Example 11>
In this example, the conditions shown in Table 12 were used using the sample (material: polystyrene, after hydrophilic treatment by plasma (water contact angle is 10° or less)) of the drop forming portion (see FIG. 9B) of the fifth embodiment. It was verified whether or not the drop could be formed, and when the drop could be formed, the influence of the difference in the diameter D on the drop was evaluated.

In Table 12, h1 indicates the distance from the upper end surface of the peripheral wall portion to the upper surface of the container bottom portion, h2 indicates the distance from the upper end surface of the peripheral wall portion to the bottom surface of the recess, and f indicates the thickness of the peripheral wall portion in the horizontal direction. From the results of Table 12, in the present embodiment, it is possible to form drops, and the larger the diameter D, the larger the capacity of the drops that can be formed, and the larger the capacity, the larger the thickness of the drops. It was confirmed that the range of formation was wide. On the other hand, in the comparison when the same amount of liquid was added, the smaller the diameter D, the thicker the drop.

<Example 12>
In this example, a sample (material: polystyrene, after hydrophilic treatment by plasma (water contact angle is 10° or less)) of the drop forming portion (see FIG. 13B) according to the modification of the fifth embodiment was used. It was verified whether or not the drop could be formed under the conditions described in 1. Note that g in Table 13 indicates the distance of the sharp portion in the vertical direction (that is, the distance from the upper end to the boundary portion between the inclined portion and the vertical portion on the outer wall surface).

As shown in Table 13, it was confirmed that the drop could be formed even if the peripheral wall portion had a sharp upper end.

<Example 13>
In this example, a sample (material: polystyrene, after hydrophilic treatment by plasma (water contact angle is 10° or less), liquid amount: 30 μL) of the drop forming portion (see FIGS. 19A to 19C) of the seventh embodiment was used. Then, vibration was applied after forming the drops, and changes in the drops before and after the vibration were evaluated. Table 14 shows the dimensions of the sample.

FIG. 30B is a photograph showing the result of Example 13. In FIG. 30B, the left side shows a photograph after the vibration is applied, and the right side shows a photograph before the vibration is applied. From the photograph, it was confirmed that the drop did not collapse even when it was vibrated and stayed on the surrounding wall.

<Example 14>
In this example, a sample (material: polystyrene, after hydrophilic treatment by plasma (water contact angle is 10° or less), liquid amount: 15 μL) of the drop forming portion (see FIGS. 20A to 20C) of the eighth embodiment was used. It was verified whether or not the drop could be formed. The dimensions of the sample are shown in Table 15.

As a result, it was confirmed that drops could be formed.

<Example 15>
In this example, a sample of the drop forming portion (see FIG. 21E) provided at the place M4 of the ninth embodiment (material: polystyrene, after hydrophilic treatment by plasma (water contact angle is 10° or less), liquid amount: (30 μL) was used to apply a vibration after forming the drop, and the change in the drop before and after the vibration was evaluated. Table 16 shows the dimensions of the sample.

As a result, it was confirmed that not only the drops were formed, but also when the drops were vibrated, the drops did not collapse and remained on the surrounding wall.

<Example 16>
In this example, by using the seven types of samples (made of polystyrene) shown in Table 17, 15 μL of water can be dropped after plasma hydrophilization treatment (water contact angle is 10° or less) to form a drop. It was verified whether or not.

As a result, any of the samples listed in Table 17 could form drops.

<Example 17>
In this example, a sample (material: polystyrene, after hydrophilic treatment by plasma, which was processed to have a curved surface portion between the upper surface and the side surface as shown in FIG. 33, was applied to the drop forming portion of the sixth embodiment. (Water contact angle is 10° or less) was used to verify whether or not a drop could be formed. The liquid volumes used were 30 μL and 50 μL. Table 18 shows the dimensions of the sample. In Table 18, R is the radius of curvature of the curved surface portion, α′ is the angle between the straight line along the upper surface and the straight line along the side surface of the drop forming portion, and d is the upper surface, as described in the thirteenth embodiment. It is the distance from the intersection P5 of the straight line along the line and the straight line along the side surface to the edge P2 of the upper surface of the container bottom adjacent via the groove.

As a result, it was confirmed that the drop can be formed even with the drop forming portion processed to have the curved surface portion.

<Example 18>
In this example, as shown in FIG. 34, a sample (material: polystyrene, hydrophilization treatment by plasma, which was processed so as to have a curved surface portion between the upper surface and the side surface with respect to the drop forming portion of the sixth embodiment described above. After that (the water contact angle is 10° or less), it was verified whether or not the drop could be formed. The liquid volume used was 15 μL. Table 19 shows the dimensions of the sample. In Table 19, R is the radius of curvature of the curved surface portion, α′ is the angle between the straight line along the upper surface and the straight line along the side surface of the drop forming portion, and d is the upper surface, as described in the thirteenth embodiment. It is the distance from the intersection P5 of the straight line along the line and the straight line along the side surface to the edge P2 of the upper surface of the container bottom adjacent via the groove.

As a result, it was confirmed that the drop can be formed even with the drop forming portion processed to have the curved surface portion.

<Example 19>
In the present example, based on the conditions shown in FIG. 35, a sample of the drop forming portion in which the upper surface and the side surface are connected via the curved surface portion (material: polystyrene, after hydrophilic treatment by plasma (water contact angle is 10° The following)) was used to verify whether or not a water drop can be formed. The liquid volumes used were 10 μL, 15 μL and 20 μL. In FIG. 35, R is the radius of curvature of the curved surface portion, and α′ is the angle formed by the straight line along the upper surface and the straight line along the side surface in the thirteenth embodiment.

As a result, as shown in FIG. 35, it was confirmed that water drops could be formed in all the samples except the case where they were not carried out.

<Example 20>
In the present example, based on the conditions shown in FIG. 36, a sample of the drop forming portion in which the upper surface and the side surface are connected via the curved surface portion (material: polystyrene, after hydrophilic treatment by plasma (water contact angle is 10°) The following)) was used to verify whether or not a culture solution drop could be formed. The liquid volumes used were 10 μL, 15 μL and 20 μL. And the culture medium used is
It was a Modified HTF Medium manufactured by Irvine Scientific. In FIG. 36, R is the radius of curvature of the curved surface portion, and α′ is the angle formed by the straight line along the upper surface and the straight line along the side surface in the thirteenth embodiment.

As a result, as shown in FIG. 36, it was confirmed that culture solution drops could be formed in all the samples except the case where the culture was not performed.

<Example 21>
38A to 38C are photographs of the cell handling container used in Example 21, FIG. 38A is a cell handling container with a lid, FIG. 38B is a cell handling container without a lid, and FIG. 38C is a plan view of a cell handling container without a lid. Is a photograph showing. The cell handling container used in this example is processed so that the drop forming portion and the surrounding wall portion each have a curved surface portion with respect to the cell handling container 60 of the twelfth embodiment, and It is processed so that the side surfaces are inclined. The specific shapes of the drop forming portion and the surrounding wall portion are as shown in FIG. In FIG. 39, R is the radius of curvature of the curved surface portion, α″ is the inclination angle of the side surface with respect to the vertical direction (that is, the vertical direction), and the angle α′ (that is, as described in the thirteenth embodiment). In addition, the relation between the straight line along the upper surface and the straight line along the side surface is α″=α′−90°. Further, D′=5 mm, d′=d″=d′″=0.4 mm, and t′=250 μm.

As shown in Table 20, 6 types of the above-mentioned cell handling containers having different radii of curvature and inclination angles were made (material: polystyrene, after hydrophilic treatment by plasma (water contact angle is 10° or less)), and dropped for each type. The forming performance was evaluated. In addition, what is shown in the photographs of FIGS. 38A to 38C is a type 6 cell handling container. The culture solution used for the evaluation was Modified HTF Medium manufactured by Irvine Scientific, and the liquid volumes were 10 μL, 15 μL, 20 μL, 25 μL and 30 μL, respectively.

Table 21 shows the evaluation results of this example. As can be seen from Table 21, in all types of cell handling containers, the drop was held at the drop forming part.

Further, in the present example, vibration was applied to the held drops, and changes in the drops were also evaluated. Table 22 shows the evaluation results after vibration. As can be seen from Table 22, when vibration was applied, it was confirmed that the drop S (see FIG. 40A) spread to the surroundings due to the vibration, but stayed in the surrounding wall portion without collapsing (see FIG. 40B). It was also confirmed that the thickness of the drop S was reduced due to the spread of the drop S.

Although the embodiments of the present invention have been described in detail above, the present invention is not limited to the above-described embodiments, and various designs are possible within the scope of the present invention as set forth in the claims. You can make changes. For example, in the above-described embodiment, the cell handling container provided with microwells has been described, but the present invention is also applied to the cell handling container not provided with microwells. The present invention is also applied to a cell handling container that does not have the lid 1b.

Further, in the above-described embodiment, the drop forming portion having a circular shape in plan view has been described, but the present invention is also applicable to a polygon such as a quadrangle in plan view, an ellipse, and the like. Furthermore, the above-described embodiments and modifications are only part of the present invention, and the present invention is also applied to each combination of the above-mentioned embodiments and modifications.

1,2,3,4,5,6,7,8,9,53,57,60,65,66 Cell handling container 10 Container bottom 10a Upper surface 11 Side wall 12 Cell accommodation area 13 Microwell 14 Culture solution accommodation (Liquid storage part)
15,16,16' Drop forming portions 15a, 16a, 16'a, 54a, 64a Upper surfaces 15b, 15c, 15d, 15e, 54b, 64c Side surfaces 16b, 16'b, 16c, 16d, 16e, 16f, 16g, 16h , 16i, 16j, 16k Side surface 17, 18, 19, 20, 20, 21, 22, 23, 24, 25, 27, 28, 43 Groove portion 26, 33, 44 Enclosure wall portion 30, 40, 54 Drop forming portion 31, 41 Recessed portion 32, 42 peripheral wall portions 32b, 32i, 32j, 42b, 42h outer wall surfaces 32c, 42c upper end surface 61 outer wall well 62 curved wall 63 recessed portion 64b curved surface portion P1 edge S drop C cell

Claims (10)

A cell handling container having a container bottom,
At least one drop forming portion having an upper surface that is a liquid drop forming region and a side surface that is provided continuously or intermittently along the circumferential direction of the upper surface and that extends downward is provided,
Between the drop forming portion and the container bottom portion, a groove portion for surrounding the drop forming portion and separating the upper surface of the drop forming portion from the upper surface of the container bottom portion is provided.
Wherein said upper surface of the drop forming portion and the upper surface of the container bottom, Ri height Do different in the vertical direction,
On the upper surface of the drop forming portion, a concave portion that opens upward is formed,
In the vertical direction, when the distance from the upper surface of the drop forming portion to the upper surface of the container bottom portion is h3, and the distance from the upper surface of the drop forming portion to the deepest portion of the recess is h4, h3+h4≦1 mm Is a cell handling container. The cell handling container according to claim 1, wherein the upper surface of the drop forming portion is a non-flat surface. In the vertical direction, the upper surface cell handling container according to claim 1 or 2 lower than the upper surface of the container bottom of the drop forming unit. The cell handling container according to any one of claims 1 to 3 , wherein h3+h4 <0.2 mm . The groove, cell handling vessel according to any one of claims 1-4 is double or more. In the vertical direction, the side surface of the drop forming unit, cell handling vessel according to any one of claims 1 to 5 which extends down to the range from the edge of the above 5μm of the upper surface of the drop forming unit. Wherein said upper surface an angle and forms the side surface of the drop forming unit, cell handling vessel according to any one of the drop claim than 136 ° or less at the edge of the upper surface of the forming portions 1-6. In the horizontal direction, the distance to the edge of the upper surface of the container bottom portion adjacent through the groove from the edge of the upper surface of the drop forming unit, any one of claim 1 to 7 at 0.01mm or more The cell handling container described in. At least the upper surface of the drop forming unit, cell handling vessel according to any one of claims 1 to 8 which is a hydrophilic surface. The cell handling container according to claim 9 , wherein a water contact angle of the hydrophilic surface is less than 80°.