Centrifuge tube, kit and method for producing microparticle gel

Abstract

To provide a centrifugal tube that can suppress generation of contamination when collecting particulate gel, and to provide a kit and a method for manufacturing particulate gel.SOLUTION: A centrifugal tube includes: a first container 20 including a first bottom 21, a first shank 22 and a first opening 23; a lid material 30 provided in the first opening of the first container so as to be capable of being removed therefrom; and a second container 40 including a second shank 41 and a second opening 42. The second opening of the second container has a width of 0.2 μm or more and 2000 μm or less. The second container is held by the lid material. The second opening of the second container is oriented on a side of the first bottom. The second opening is communicated to the interior of the first container.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present disclosure relates to a centrifuge tube, a kit, and a method for producing a microparticle gel. [Background technology]

[0002] 2. Description of the Related Art Conventionally, as disclosed in Patent Document 1, for example, there is known a technique for producing a particulate gel by dripping a gelling solution using centrifugal force.

[0003] However, in the device described in Patent Document 1, the tubular part for dropping is held in the gelling container. Therefore, when recovering the produced microparticle gel, the tubular part may be an obstacle, making it difficult to recover the microparticle gel. In addition, it is necessary to carefully remove the tubular part with sterilized tweezers or the like before recovering the microparticle gel, and there is a risk that a complicated operation will be required to recover the microparticle gel without causing contamination. In addition, since the tubular part is held in the gelling container, friction occurs at the part that holds the tubular part, which may cause foreign matter to be generated in the gelling container, which may also be a risk of causing contamination.

[0004] Furthermore, for example, Patent Document 2 discloses a technique for producing gel particles by ejecting a gelling solution in the form of droplets using centrifugal force.

[0005] However, in the device described in Patent Document 2, the container containing the gelling solution and the container containing the microparticle gel are not integrated. Therefore, when applying centrifugal force to the container, only the container containing the gelling solution is rotated. In this case, the gelling solution may scatter around, which may result in the device becoming larger and the risk of contamination increasing. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] JP 2012-176374 A [Patent Document 2] JP 2013-158753 A Summary of the Invention [Problem to be solved by the invention]

[0007] The present disclosure provides a centrifuge tube, a kit, and a method for producing a microparticle gel, which are capable of suppressing the occurrence of contamination when recovering the microparticle gel. [Means for solving the problem]

[0008] The centrifuge tube according to this embodiment comprises a first container having a first bottom, a first body, and a first opening, a lid material removably attached to the first opening of the first container, and a second container having a second body and a second opening, wherein the width of the second opening of the second container is 0.2 μm or more and 2000 μm or less, the second container is held by the lid material, the second opening of the second container faces the first bottom side, and the second opening is connected to the interior of the first container.

[0009] In the centrifuge tube according to this embodiment, the first body portion and the cover member may be fitted or threadedly engaged with each other.

[0010] In the centrifuge tube of this embodiment, the lid material has a lid top surface portion that covers the first opening of the first container, a lid opening is formed in the lid top surface portion, and the second container may be positioned within the lid opening.

[0011] In the centrifuge tube of this embodiment, a communication portion may be provided between the first container and the lid, or between the second container and the lid, which communicates between the space between the first container and the second container and the outside air.

[0012] In the centrifuge tube according to the present embodiment, a droplet guiding portion that guides droplets from the second opening of the second container may be provided in the first container.

[0013] In the centrifuge tube according to the present embodiment, a plurality of the second containers may be provided.

[0014] The kit according to this embodiment is a kit for producing a centrifuge tube according to this embodiment, and includes a first container having a first bottom, a first body, and a first opening, a lid material removably attached to the first opening of the first container, and a second container having a second body and a second opening.

[0015] The method for producing a particulate gel according to this embodiment includes the steps of preparing a centrifuge tube according to this embodiment, storing a first gelling solution in the first container and a second gelling solution in the second container, and applying centrifugal force to the centrifuge tube to drip droplets of the second gelling solution from the second container into the first gelling solution stored in the first container.

[0016] The method for producing a particulate gel according to this embodiment includes the steps of preparing a centrifuge tube according to this embodiment, storing a first gelling solution in one of the multiple second containers and storing a second gelling solution in the other second containers, and applying centrifugal force to the centrifuge tube to drip droplets of the first gelling solution from the second container containing the first gelling solution into the first container and drip droplets of the second gelling solution from the second container containing the second gelling solution into the first container. Effect of the Invention

[0017] According to this embodiment, it is possible to suppress the occurrence of contamination when collecting the fine particle gel. [Brief description of the drawings]

[0018] [Figure 1] FIG. 1 is a vertical cross-sectional view showing a centrifuge tube according to one embodiment. [Diagram 2] FIG. 2 is a vertical sectional view showing a centrifuge tube according to a modified example. [Diagram 3] 3(a)-(d) are diagrams showing a method of using a centrifuge tube according to one embodiment. [Figure 4] FIG. 4 is a vertical sectional view showing a centrifuge tube according to a first modified example. [Diagram 5] FIG. 5 is a vertical sectional view showing a centrifuge tube according to a second modified example. [Figure 6] FIG. 6 is a vertical sectional view showing another configuration of the centrifuge tube according to the second modified example. [Figure 7] FIG. 7 is an optical microscope photograph showing a particulate gel produced using the centrifuge tube of Example 1. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0019] Hereinafter, an embodiment will be specifically described with reference to the drawings. Each of the figures shown below is a schematic illustration. Therefore, the size and shape of each part are appropriately exaggerated to facilitate understanding. In addition, it is possible to carry out appropriate modifications within the scope of the technical idea. In each of the figures shown below, the same parts are given the same reference numerals, and some detailed explanations may be omitted. In addition, the numerical values ​​such as the dimensions of each member and the material names described in this specification are examples of embodiments, and are not limited to these, and can be appropriately selected and used. In this specification, terms that specify shapes and geometric conditions, such as parallel, orthogonal, and vertical, are intended to include substantially the same state in addition to their strict meanings. In addition, for convenience of explanation, the terms "upper" and "lower" may be used in the explanation, but the up-down direction may be reversed.

[0020] In this specification, "upward" refers to the direction of the central axis of the centrifuge tube that faces the side opposite to the first bottom of the first container, and "downward" refers to the direction of the central axis of the centrifuge tube that faces the first bottom of the first container.

[0021] The present embodiment will be described below, but the present embodiment is not limited to the following description.

[0022] [Centrifuge tube] First, the configuration of a centrifuge tube according to the present embodiment will be described with reference to Fig. 1. Fig. 1 is a cross-sectional view showing a centrifuge tube 10 according to the present embodiment.

[0023] As shown in Fig. 1, the centrifuge tube 10 is an instrument for dripping a solution by centrifugal force to produce a microparticle gel with high size uniformity. This centrifuge tube 10 is suitable for producing microparticles in fields that require sterility and reduce the risk of contamination from the external environment, such as producing a microparticle gel as a cell culture carrier for culturing cells on the surface and inside, or a microparticle gel for use in a drug delivery system. In addition, the centrifuge tube 10 is particularly suitable for use in the reaction of droplets of a sodium alginate solution with a calcium chloride solution to produce a calcium alginate microparticle gel.

[0024] The centrifuge tube 10 according to the present embodiment includes a first container 20, a cover member 30, and a second container 40. The first container 20 includes a first bottom 21, a first body 22, and a first opening 23. The cover member 30 is removably attached to the first opening 23 of the first container 20. The second container 40 includes a second body 41 and a second opening 42, and the second container 40 is held by the cover member 30. The width W1 of the second opening 42 of the second container 40 is 0.2 μm or more and 2000 μm or less. The second opening 42 of the second container 40 faces the first bottom 21 side, and the second opening 42 communicates with the inside of the first container 20. In this case, the centrifuge tube 10 has a central axis CL.

[0025] [First container] The first container 20 is a container for accommodating a solution such as a first gelling solution, and has a first bottom 21 located at the bottom, a first body 22 connected to the first bottom 21, and a first opening 23 located above the first body 22. Among these, the first bottom 21 and the first body 22 form a space for accommodating the solution. The first opening 23 forms an opening for filling the first container 20 with the solution or removing the solution from the first container 20.

[0026] The configuration of the first body 22 around the first opening 23 is not particularly limited as long as the lid 30 can be attached and detached. For example, the first body 22 and the lid 30 may be shaped to fit together, or the first body 22 and the lid 30 may be structured to be screw-engaged with each other.

[0027] The structure of the first bottom 21 is not particularly limited, and may be, for example, a flat bottom, an inclined bottom, a V-shaped bottom, a U-shaped bottom, or the like. In particular, it is preferable that the first container 20 has an inclined portion 24 where the microparticle gel accumulates at a predetermined position so that the microparticle gel formed in the first container 20 can be easily collected. The lowest point where the microparticle gel accumulates is not particularly limited, and may be the center position of the first bottom 21 (a position passing through the central axis CL) or an end portion shifted from the center position of the first bottom 21. It is more preferable that the lowest point is shifted from directly below the second opening 42 of the second container 40. The reason for this is as follows. The produced microparticle gel accumulates upward from the lowest point position, but if the microparticle gel reaches near the liquid surface of the first gelling solution, the microparticle gel becomes an obstacle and the first gelling solution is difficult to be appropriately dropped into the second gelling solution, which may make it difficult to produce a uniform microparticle gel. For this reason, the further the lowest point position is shifted from directly below the second opening 42, the less likely the microparticle gel will hinder the dripping, making it easier to produce uniform microparticle gel.

[0028] In addition, the first body 22 is preferably made of a transparent material so that the amount of the solution can be easily visually checked from the outside. It is more preferable that the first body 22 has a scale printed thereon.

[0029] The material of the first container 20 is not particularly limited as long as it can contain the solution so as to reduce the risk of contamination from the outside, and suitable materials include polyolefin resins such as polyethylene and polypropylene, polyvinyl chloride, ethylene-vinyl acetate copolymer resin, C-FLEX, polystyrene, polyethylene terephthalate, glass, silicone resin, fluororesin, acrylic resin, urethane resin, etc. In addition, it is desirable that the material of the first container 20 is a hard material so as not to be deformed by the centrifugal force applied to the first container 20.

[0030] [Lid material] As described above, the lid material 30 is removably provided on the first opening 23 of the first container 20. The lid material 30 has a lid top surface portion 31 that covers the first opening 23 of the first container 20, and a lid peripheral portion 32 that is provided on the periphery of the lid top surface portion 31 and extends toward the first bottom portion 21. A lid opening 33 is formed in the lid top surface portion 31, and the second container 40 is located within the lid opening 33. The lid material 30 may have a portion that does not cover the first opening 23 of the first container 20, but it is preferable that the uncovered portion is as small as possible. In addition, the lid material 30 and the second container 40 held by the lid material 30 may be partially present inside the first container 20, but it is preferable that at least a portion of the lid material 30 and the second container 40 are located above the first opening 23 of the first container 20. This makes it possible to remove the lid material 30 and the second container 40 from the first container 20 by gripping the portion of the lid material 30 or the second container 40 located above when recovering the microparticle gel. This eliminates the need to carefully insert sterilized tweezers or the like into the first container 20, making it possible to collect the microparticle gel with less risk of contamination.

[0031] The material of the lid member 30 is not particularly limited, and suitable materials include polyolefin resins such as polyethylene and polypropylene, polyvinyl chloride, ethylene-vinyl acetate copolymer resin, C-FLEX, polystyrene, polyethylene terephthalate, glass, silicone resin, fluororesin, acrylic resin, urethane resin, etc. The material of the lid member 30 may be the same as or different from the material of the first container 20 and / or the second container 40.

[0032] [Second container] The second container 40 is a container for accommodating a solution such as a second gelling solution, and has a second body 41 and a second opening 42 located below the second body 41. The second body 41 is made of a substantially cylindrical member. The second opening 42 faces the first bottom 21, and when viewed from the second opening 42 side, a part of the inside of the first container 20 can be confirmed. The second opening 42 is also open toward the inside of the first container 20 and communicates with the internal space of the first container 20. The second opening 42 is located closer to the first bottom 21 of the first container 20 than the lid 30 and faces the first bottom 21 side. Note that, as long as a part of the inside of the first container 20 can be confirmed when viewed from the second opening 42 side, the second opening 42 may be located on the opposite side of the first bottom 21 with respect to the lid 30. The width W1 of the second opening 42 is narrowed, specifically, 0.2 μm or more and 2000 μm or less. For this reason, under normal circumstances (when no force such as centrifugal force is applied), the solution contained in the second body 41 does not drip from the second opening 42. On the other hand, when centrifugal force is applied to the solution in the second container 40, the solution drips from the second opening 42 towards the first container 20. The second opening 42 may be a hole formed in the bottom of the second container 40, and more preferably has a nozzle-like structure protruding downward from the bottom of the second container 40 so that the solution can be easily dripped.

[0033] A tapered portion 43 is preferably provided at the bottom of the second body 41. This allows the second gelling solution or other solution to be discharged from the second opening 42 with as little waste as possible. The shape and size of the second opening 42 can be appropriately selected, and this allows the size of the droplets of the solution to be dropped from the second opening 42 to be adjusted. For example, the shape of the second opening 42 is preferably a circle or a regular polygon. The width W1 of the second opening 42 is 0.2 μm or more and 2000 μm or less, preferably 1 μm or more and 1000 μm or less, and more preferably 10 μm or more and 200 μm or less. In particular, when cells are to be supported on the prepared microparticle gel, the larger the width W1 of the second opening 42, the easier it is to support the cells. Desirably, when the width W1 of the second opening 42 is greater than 10 μm, a microparticle gel larger than 10 μm, which is the approximate size of a cell, can be prepared, which has the advantage of making it easier to support the cells. In addition, when the microparticle gel carrying cells is cultured by suspending it in a rotation culture or the like, the smaller the width W1 of the second opening 42, the easier it is to suspend the prepared microparticle gel. Here, the width W1 of the second opening 42 is the distance of the second opening 42 measured on a plane perpendicular to the central axis CL of the centrifuge tube 10, and refers to the length of the longest part on the plane. For example, when the shape of the second opening 42 is circular, the width W1 of the second opening 42 corresponds to the diameter of the second opening 42. Also, for example, when the shape of the second opening 42 is square, the width W1 of the second opening 42 corresponds to the length of the diagonal line of the second opening 42.

[0034] There may be a plurality of second openings 42. The presence of a plurality of second openings 42 increases the amount of droplets that can be dropped in a certain time, which is more desirable from the viewpoint of throughput. In this case, if the interval between the second openings 42 is narrow, the dropped solution is likely to fuse near the second openings 42, and the dropped solution becomes larger than expected. For this reason, the solution is likely to be dropped without centrifugation, which may make it difficult to control the dropping or to prepare a uniform microparticle gel. In addition, when the solutions dropped from the respective second openings 42 fuse together to gel, the microparticle gel adheres to the second openings 42 and prevents the dropping, so the greater the interval between the second openings 42, the more preferable it is. Specifically, the interval between the second openings 42 is preferably 1 time or more the width W1 of the second openings 42, more preferably 10 times or more, and even more preferably 100 times or more. In addition, when three or more second openings 42 are provided, it is preferable that the interval between all the second openings 42 and all the other second openings 42 is within the above range. In addition, when the interval between the second openings 42 is smaller than 1 time the width W1 of the second openings 42, it may be assumed that the dropped solution will necessarily fuse near the second openings 42 before being dropped. In this case, the sum of the widths W1 of the multiple second openings 42 adjacent to each other at narrow intervals as described above is preferably 0.2 μm to 2000 μm, more preferably 1 μm to 1000 μm, and even more preferably 10 μm to 200 μm. This makes it difficult for the solution to be dropped when not centrifuged, as in the case where the dropped solution does not fuse, and also makes it easier to support cells on the prepared microparticle gel and to perform rotational culture.

[0035] The means for holding the second container 40 on the lid member 30 is not limited. That is, it is sufficient that the second container 40 does not come off the lid member 30 when applying centrifugal force to the centrifuge tube 10. In FIG. 1, the second container 40 is penetrated through the lid opening 33 of the lid member 30, and the upper part of the second container 40 is hooked on the lid member 30. Alternatively, the second container 40 and the lid member 30 may be fixed with an adhesive or the like. The second container 40 and the lid member 30 may be integrally formed. Alternatively, at the connection part between the first container 20 and the lid member 30, a part of the second container 40 may be sandwiched and fixed between the first container 20 and the lid member 30. Furthermore, the second container 40 and the lid member 30 may not be completely fixed, and the second container 40 and the lid member 30 may be movably connected so that the position of the second container 40 can be adjusted up, down, left, and right to control the dropping conditions. The position where the second container 40 is held is not particularly limited, and may be the center of the lid member 30 or the end of the lid member 30.

[0036] The second container 40 further has a third opening 44 located above the second body 41. The third opening 44 faces the opposite side to the second opening 42, and is an opening for adding a solution into the second container 40. A flange portion 45 for holding the second container 40 to the lid 30 is formed around the third opening 44. Note that, in order to prevent contamination from the third opening 44, it is desirable to further provide a lid (not shown) that covers the third opening 44.

[0037] The material of the second container 40 is preferably transparent so that the amount of liquid in the second container 40 can be easily visually observed from the outside. It is more preferable that a scale is printed on the second body 41. The material of the second container 40 is preferably a polyolefin resin such as polyethylene or polypropylene, polyvinyl chloride, ethylene-vinyl acetate copolymer resin, C-FLEX, polystyrene, polyethylene terephthalate, glass, silicon resin, fluororesin, acrylic resin, urethane resin, or the like. The material of the second container 40 is preferably a hard material so as not to be deformed by the centrifugal force applied to the second container 40.

[0038] As shown in FIG. 2, a plurality of second containers 40 may be provided. In this case, the same type of solution may be contained in all of the plurality of second containers 40, or different types of solutions may be contained in the plurality of second containers 40. When the same type of solution is contained in the plurality of second containers 40, the amount of droplets that can be dropped in a certain time can be increased by dropping the solutions from the plurality of second containers 40 at the same time, which is more desirable from the viewpoint of throughput. In this case, the interval between the second opening 42 of the second container 40 and the second opening 42 of the second container 40 different from the second opening 42 is preferably 1 time or more the width W1 of the second opening 42, more preferably 10 times or more, and even more preferably 100 times or more. By setting the interval between the second openings 42 to the above range, it is possible to suppress the tendency of the dropped solution to fuse near the second openings 42, and to prevent the dropped solution from becoming larger than expected. Therefore, it is possible to suppress the solution from dropping without centrifugation, and to suppress the difficulty of controlling the dropping and the difficulty of producing a uniform microparticle gel. Furthermore, in the case where the solutions dropped from the respective second openings 42 are fused together to gel, it is possible to prevent the particulate gel from adhering to the second openings 42 or from interfering with the dropping. In the case where different types of solutions are contained in the multiple second containers 40, at least one second container 40 may contain the second gelling solution, and the other second containers 40 may contain a cell suspension, a drug, the first gelling solution, or the like.

[0039] [Whole centrifuge tube] The centrifuge tube 10 preferably has excellent centrifugal strength. Excellent centrifugal strength means, for example, that the tube has a strength that does not break or crack in a centrifuge test at 1500 rpm. More preferably, the centrifuge tube 10 does not break or crack in a centrifuge test at 3000 rpm, and even more preferably, in a centrifuge test at 4000 rpm. An example of the evaluation conditions for the centrifuge test is as follows: Centrifuge: HITACH CF16RX, rotor used: swing rotor, adapter compatible with 50 mL centrifuge tube, set centrifugal strength: 1500 rpm or more, set temperature: 20°C, time: 10 minutes, content: 10 mL or more of pure water.

[0040] [Gelling solution] The first gelling solution contained in the first container 20 and the second container 40 are not particularly limited as long as they are mixed together to gel. For example, an aqueous solution of a high-value cationic salt such as calcium chloride is preferably used as the first gelling solution, and a sodium alginate solution is preferably used as the second gelling solution. When preparing a particulate gel containing a substance such as cells or a drug, it is desirable to have the substance contained in the first gelling solution or the second gelling solution. The second gelling solution contained in the second container 40 may further contain a substance capable of adjusting the surface tension, such as a surfactant, or a substance capable of adjusting the viscosity, such as a thickener, in order to control the dropping.

[0041] [Operation of this embodiment] Next, the operation of the present embodiment having the above-mentioned configuration will be described. Specifically, a method for producing a particulate gel using the above-mentioned centrifuge tube 10 will be described.

[0042] First, the above-mentioned centrifuge tube 10 is prepared as shown in Fig. 3(a). The centrifuge tube 10 may be a kit in which each component (e.g., the first container 20, the cover member 30, and the second container 40) is separately configured and can be assembled to form the structure of the centrifuge tube 10 when necessary, in which case the centrifuge tube 10 is assembled before use. In this embodiment, a kit for preparing such a centrifuge tube is also provided.

[0043] Next, as shown in FIG. 3(b), the first gelling solution L1 is placed in the first container 20 of the centrifuge tube 10, and the second gelling solution L2 is placed in the second container 40.

[0044] Next, as shown in FIG. 3(c), centrifugal force is applied to the centrifuge tube 10 (see the arrow in FIG. 3(c)). This centrifugal force pushes the second gelling solution L2 out and passes through the second opening 42 having a narrow width W1. This allows droplets of the second gelling solution L2 to be dropped from the second opening 42 into the first gelling solution L1 contained in the first container 20. The droplets of the second gelling solution L2 undergo a gelling reaction with the first gelling solution L1 in the first container 20 to generate a particulate gel. In this case, the generated particulate gel can be accumulated on the first bottom 21 of the first container 20.

[0045] When applying centrifugal force to the centrifuge tube 10, it is preferable to place the centrifuge tube 10 in a centrifuge (not shown) and centrifuge it. The rotor of the centrifuge may be an angle rotor or a swing rotor. For example, it is preferable to use a swing rotor to apply centrifugal force directly below the centrifuge tube 10, and it is preferable to use an angle rotor to apply centrifugal force diagonally below the centrifuge tube 10. The direction in which the centrifugal force is applied can be controlled by adjusting the installation angle of the centrifuge tube 10 and the balance of the center of gravity position, in addition to selecting the rotor described above.

[0046] 3(d), after the dripping of the second gelling solution L2 is completed, the second container 40 is removed from the first container 20 together with the lid 30. Then, the solution containing the microparticle gel is taken out from the first container 20 using a pipette or the like (not shown).

[0047] In this embodiment, the second container 40 is held by the lid 30. As a result, after the microparticle gel is produced, the second container 40 can be easily removed from the first container 20 by removing the lid 30. In addition, the lid 30 for attaching the second container 40 to the first container 20 is not present inside the first container 20. Therefore, the generation of foreign matter caused by the lid 30 rubbing against the first container 20 inside the first container 20 is suppressed, and the risk of contamination can be reduced. As a result, the solution containing the microparticle gel can be easily collected from the first container 20 with a pipette or the like with little risk of contamination. In addition, since the second container 40 and the first container 20 are connected by the lid 30, the centrifuge tube 10 can be easily configured.

[0048] In addition, when a plurality of second containers 40 are provided as shown in FIG. 2, one of the plurality of second containers 40 may contain the first gelling solution L1, and the other second containers 40 may contain the second gelling solution L2. In this case, by applying centrifugal force to the centrifuge tube 10, droplets of the first gelling solution L1 are dropped from the second container 40 containing the first gelling solution L1 into the first container 20. Also, droplets of the second gelling solution L2 are dropped from the second container 40 containing the second gelling solution L2 into the first container 20. In this way, the droplets of the first gelling solution L1 and the droplets of the second gelling solution L2 dropped from the plurality of second containers 40 may be fused in the first container 20 to produce a microparticle gel.

[0049] As described above, according to this embodiment, the width W1 of the second opening 42 of the second container 40 is 0.2 μm or more and 2000 μm or less, and the second container 40 is held by the lid material 30. As a result, by applying centrifugal force to the centrifuge tube 10, droplets of the second gelling solution L2 are dropped from the second container 40, and a microparticle gel with a high uniformity in size can be generated in the first container 20. In addition, by removing the removable lid material 30 from the first container 20, the second container 40 held by the lid material 30 can also be easily removed. As a result, the microparticle gel can be collected from the first container 20 easily and with a reduced risk of contamination.

[0050] [Variations] Next, each modified example of the present disclosure will be described with reference to Fig. 4 to Fig. 6. Fig. 4 to Fig. 6 are views showing a centrifuge tube according to a modified example of the present disclosure. In Fig. 4 to Fig. 6, the same parts as those shown in Fig. 1 to Fig. 3 are given the same reference numerals and detailed description will be omitted.

[0051] (First Modification) Fig. 4 shows a vertical cross section of a centrifuge tube 10 according to a first modified example. In Fig. 4, a first communication part 25 is formed between the first container 20 and the cover member 30. In addition, a second communication part 34 is formed between the second container 40 and the cover member 30. The first communication part 25 and the second communication part 34 each communicate the space S between the first container 20 and the second container 40 with the outside air. Only one of the first communication part 25 and the second communication part 34 may be provided, or both may be provided.

[0052] The first communication part 25 is a connection part between the first container 20 and the lid material 30, and is provided on the first container 20 side. That is, the first container 20 has the first communication part 25. However, the present invention is not limited to this, and the first communication part 25 may be provided on the lid material 30 side. That is, the lid material 30 may have the first communication part 25. Alternatively, the first communication part 25 may be a gap that occurs between the first container 20 and the lid material 30 when the first container 20 and the lid material 30 are fitted together. The first communication part 25 may have a structure in which a gap is formed without completely blocking the connection part between the first container 20 and the lid material 30 by a protrusion or the like. Alternatively, the first communication part 25 may be formed by providing a through hole at a position of the first container 20 and the lid material 30 where there is no problem in containing the solution.

[0053] The second communication part 34 is a connection part between the second container 40 and the lid material 30, and is provided on the lid material 30 side. That is, the lid material 30 has the second communication part 34. However, the present invention is not limited to this, and the second communication part 34 may be provided on the second container 40 side. That is, the second container 40 may have the second communication part 34. Alternatively, the second communication part 34 may be a gap that occurs between the second container 40 and the lid material 30 when the second container 40 and the lid material 30 are fitted together. The second communication part 34 may have a structure in which a gap is formed without completely blocking the connection part between the second container 40 and the lid material 30 by a protrusion or the like. Alternatively, the second communication part 34 may be formed by providing a through hole at a position of the second container 40 and the lid material 30 where there is no problem in containing the solution.

[0054] According to this modification, even if the space S between the first container 20 and the second container 40 becomes positive pressure by dripping the second gelling solution from the second container 40 into the first container 20, this positive pressure can be prevented from interfering with the dripping of the second gelling solution. For this reason, the first communication part 25 and the second communication part 34 are configured so that the gases inside and outside the space S communicate with each other and the positive pressure can be released.

[0055] The first communicating part 25 and / or the second communicating part 34 may have a valve structure that is not in communication when the second gelling solution is not dripped from the second container 40, and is in communication when the second gelling solution is dripped by centrifugal load and the space S becomes positive pressure. Also, the first communicating part 25 and / or the second communicating part 34 may be formed just before applying centrifugal force. For example, when the cover material 30 and the first container 20 and / or the second container 40 are connected by a screw type, the first communicating part 25 and / or the second communicating part 34 may be formed by loosening the screw just before applying centrifugal force to the centrifuge tube 10. Also, the first communicating part 25 and / or the second communicating part 34 may be closed with a cap (not shown) and opened by removing the cap just before applying centrifugal force to the centrifuge tube 10. In this way, by keeping the first communication portion 25 and / or the second communication portion 34 closed when not necessary, the risk of foreign matter entering the interior of the first container 20 through the first communication portion 25 and / or the second communication portion 34 can be reduced.

[0056] Furthermore, a plurality of first communication parts 25 and / or second communication parts 34 may be provided. The cross-sectional area of ​​the first communication part 25 and / or second communication part 34 is desirably larger than the cross-sectional area of ​​the second opening 42 of the second container 40 in order to facilitate the discharge of gas that has become positive pressure. Here, when a plurality of first communication parts 25 and / or second communication parts 34 are provided, the cross-sectional area is defined as the total cross-sectional area of ​​all of these.

[0057] In order to prevent foreign matter such as suspended particles from settling inside the first container 20 through the first communicating part 25 and / or the second communicating part 34, it is desirable that the outlets of the first communicating part 25 and / or the second communicating part 34 to the outside face in a direction other than the upward direction, such as a side direction or a downward direction of the centrifuge tube 10. This makes it possible to prevent foreign matter from settling inside the first container 20 through the first communicating part 25 and / or the second communicating part 34. For example, in FIG. 4, the outlets of the first communicating part 25 and the outlets of the second communicating part 34 face in a direction other than the upward direction (side direction). Note that even if the outlets of the first communicating part 25 and / or the second communicating part 34 face in the upward direction, it is preferable that the path of the first communicating part 25 and / or the second communicating part 34 is curved midway. In other words, as long as the cross section of any one of the paths of the first communicating portion 25 and / or the second communicating portion 34 does not face upward, even if a foreign object enters the first communicating portion 25 and / or the second communicating portion 34, the foreign object can be prevented from settling inside the first container 20.

[0058] (Second Modification) Fig. 5 shows a centrifuge tube 10 according to a second modified example. In Fig. 5, a droplet guiding section 50 that guides droplets from the second opening 42 of the second container 40 is provided in the first container 20. In this case, the droplet guiding section 50 may be an inclined plate that is inclined with respect to the central axis CL of the centrifuge tube 10. At least a part of the droplet guiding section 50 is provided on the first bottom 21 side of the second opening 42 and at a position spaced apart from the second opening 42.

[0059] However, if the second opening 42 comes into contact with the first gelling solution in the first container 20, the second opening 42 may become clogged and the droplets may not be dropped. Also, if the second opening 42 is completely immersed in the first gelling solution in the first container 20, the second gelling solution may be discharged from the second opening 42 in strings rather than droplets, which may hinder proper dropping. For this reason, it is considered desirable to separate the second opening 42 from the liquid surface of the first gelling solution. On the other hand, if the distance between the second opening 42 and the liquid surface of the first gelling solution is too large, the impact of the droplets dropped from the second opening 42 falling into the first gelling solution may become large, and the droplets may be deformed or broken.

[0060] In contrast to this, in this modified example, a droplet guiding section 50 that guides droplets from the second opening 42 of the second container 40 is provided in the first container 20. The droplet guiding section 50 is disposed obliquely with respect to the centrifugal force direction (the direction of the arrow in FIG. 5). This allows the droplets dropped from the second opening 42 to fall into the first gelling solution via the droplet guiding section 50, and the impact of the droplets falling into the first gelling solution can be softened. In addition, the risk of the second opening 42 of the second container 40 coming into contact with the first gelling solution can be reduced.

[0061] The position and orientation of the droplet guiding part 50 are not limited to those described above. The droplet guiding part 50 may or may not be fixed to the first container 20. The droplet guiding part 50 is preferably structured such that the droplets move obliquely to the direction of centrifugal force when centrifugal force is applied. In order to reduce the impact of the droplets dropped from the second opening 42 falling into the droplet guiding part 50, it is desirable that the distance between the droplet guiding part 50 and the second opening 42 is short, and it is more desirable that the droplet guiding part 50 and the second opening 42 are in contact with each other.

[0062] The widthwise cross-sectional shape of the droplet guiding section 50 (cross-sectional shape cut by a plane P perpendicular to the lengthwise direction (arrow A direction in FIG. 5) of the droplet guiding section 50) is desirably a shape that can appropriately guide the droplets into the first gelling solution. For example, the droplet guiding section 50 is desirably in the form of a slide, and the widthwise center is lower than the widthwise ends. Specifically, the widthwise cross-sectional shape of the droplet guiding section 50 is desirably, for example, U-shaped or V-shaped. This allows the droplets to pass through the widthwise center of the droplet guiding section 50 and fall into the first gelling solution, preventing the droplets from flying out of the droplet guiding section 50 and improving reproducibility when producing microparticle gel. In addition, the widthwise center of the droplet guiding section 50 being lower than the widthwise ends is also desirable when a plurality of second containers 40 are provided and droplets dropped from each of the second containers 40 are fused (see FIG. 2). In this case, the droplets gather at the widthwise center of the droplet guiding section 50, making it easier to fuse the droplets together.

[0063] The surface of the liquid droplet guiding portion 50 is preferably highly hydrophobic so that the liquid droplets can slide easily. The contact angle of the surface of the liquid droplet guiding portion 50 with water is preferably 60 degrees or more, more preferably 90 degrees or more, and even more preferably 120 degrees or more.

[0064] The material of the droplet guiding part 50 is preferably the same as that of the first container 20 or a material softer than that of the first container 20 in order to reduce the risk of foreign matter being generated due to friction with the inside of the first container 20. Also, in order to prevent the droplet guiding part 50 from interfering with the recovery of the microparticle gel in the first container 20, it is preferable that the microparticle gel can be recovered with a pipette or the like without removing the droplet guiding part 50. Specifically, it is preferable that the shape and arrangement of the droplet guiding part 50 are such that the tip of the pipette can approach the first bottom 21 of the first container 20 while avoiding the droplet guiding part 50.

[0065] As shown in FIG. 6, the droplet guide section 50 may be formed from the inner surface of the first body 22 of the first container 20. In this case, by applying centrifugal force in a diagonally downward direction (arrow direction in FIG. 6) with respect to the central axis CL of the centrifuge tube 10, the droplets dropped from the second opening 42 are first dropped onto the inner surface of the first body 22 of the first container 20. Then, the droplets can be dropped into the first gelling solution through the inner surface of the first body 22. In this case, the position of the second opening 42 is deviated from the central axis CL of the centrifuge tube 10, and is desirably disposed near any part of the inner surface of the first body 22. Specifically, the second container 40 may be held at a position deviated from the center of the lid member 30, the second container 40 may be held at an end of the lid member 30, or the second container 40 may be held at an angle to the lid member 30. In this way, the second opening 42 may be brought close to the inner surface of the first body 22. Furthermore, when centrifugal force is applied in an obliquely downward direction relative to the central axis CL of the centrifuge tube 10, when the centrifugal force is removed, gravity causes a flow of the first gelling solution in the opposite direction to the centrifugal force, and the resulting impact may cause the first gelling solution to come into contact with the second opening 42. In order to reduce such a risk, it is desirable that the position of the second opening 42 is shifted from the central axis CL of the centrifuge tube 10 and disposed near any part of the inner surface of the first body 22.

[0066] [Example] Next, a specific example of this embodiment will be described.

[0067] Example 1 The centrifuge tube shown in FIG. 1 was prepared. In this case, a 50 ml conical tube (product number 352070) manufactured by FALCON was prepared as the first container and the lid. A 2.5 ml syringe (product number SS-02LZ) manufactured by Terumo was prepared as the second container. Next, a through hole was opened in the center of the lid, and the second container was inserted through the through hole, and the flange of the second container was attached so that it was caught and held by the lid. At this time, it was confirmed that even if the plunger of the second container was pressed to send air into the first container, the positive pressure in the first container was immediately released. A metal needle (32G, product number SNA-32G-B) manufactured by Musashi Engineering was attached to the second container to form a second opening (inner diameter 100 μm). 12.5 ml of 1 w% calcium chloride aqueous solution was injected into the first container, the plunger of the second container was removed, and 1 ml of 1 w% sodium alginate aqueous solution was injected into the second container. Here, the distance between the second opening of the second container and the liquid surface of the 1w% calcium chloride aqueous solution was about 5 mm. Next, the centrifuge tube was attached to a Hitachi centrifuge (RXII), and a centrifugal force of 410G was applied directly below the centrifuge tube for 15 minutes. As a result, almost all of the sodium alginate solution was dripped. When measured by optical microscope observation, a highly uniform particle gel with a diameter of about 200 μm was formed at the first bottom of the first container (see FIG. 7). After that, the lid material and the second container were removed with bare hands, and the particle gel could be collected with a pipette. In addition, a lid material with a through hole in the center in advance was produced by a 3D printer without drilling a through hole as the lid material, and the same test as above was performed, and the same results were obtained.

[0068] Example 2 A centrifuge tube having a droplet guide portion as shown in FIG. 5 was produced. In this case, the centrifuge tube was produced in a manner substantially similar to that of Example 1. On the other hand, a rectangular plate made of polypropylene with a length of 65 mm and a width of 5 mm was prepared as the droplet guide portion, and this plate was placed at an angle in the first container. At this time, the distance between the second opening and the droplet guide portion was about 5 mm. Next, 5 ml of a 1 w% calcium chloride aqueous solution was poured into the first container, the plunger of the second container was removed, and 1 ml of a 1 w% sodium alginate aqueous solution was poured into the second container. In this state, as in Example 1, a centrifugal force of 410 G was applied directly below the centrifuge tube for 15 minutes using a centrifuge. As a result, almost all of the sodium alginate solution was dripped, and a highly uniform particle gel with a diameter of about 200 μm was formed at the first bottom of the first container. At this time, no dispersion of fibrous material was observed as if the particle gel had collapsed. Furthermore, when collecting the particle gel, almost all of the solution in the first container could be collected with a pipette without removing the droplet guide portion.

[0069] It is also possible to combine the multiple components disclosed in the above embodiment and modified examples as necessary. Alternatively, some components may be deleted from all the components shown in the above embodiment and modified examples. [Explanation of symbols]

[0070] 10 Centrifuge tubes 20 First container 21 First bottom 22 First body 23 First opening 24 Slope 25 First communication section 30 Lid material 31 Lid top section 32 Lid rim 33 Lid opening 34 Second communication part 40 Second container 41 Second body 42 Second opening 43 Tapered section 44 Third opening 45 Flange part 50 Droplet induction part

Claims

1. a first container having a first base, a first body, and a first opening; A cover material removably provided on the first opening of the first container; a second container having a second body and a second opening; The width of the second opening of the second container is 0.2 μm or more and 2000 μm or less, The second container is held by the lid, The second opening of the second container faces the first bottom side, The second opening communicates with the interior of the first container, A centrifuge tube, wherein a communication portion is provided between the first container and the lid, or between the second container and the lid, for communicating the space between the first container and the second container with the outside air.

2. A first container having a first bottom, a first body, and a first opening; A cover material removably provided on the first opening of the first container; a second container having a second body and a second opening; The width of the second opening of the second container is 0.2 μm or more and 2000 μm or less, The second container is held by the lid, The second opening of the second container faces the first bottom side, The second opening communicates with the interior of the first container, A centrifuge tube, the first container being provided with a droplet guide portion configured to guide droplets from the second opening of the second container.

3. A first container having a first bottom, a first body, and a first opening; A cover material removably provided on the first opening of the first container; a second container having a second body and a second opening; The width of the second opening of the second container is 0.2 μm or more and 2000 μm or less, The second container is held by the lid, The second opening of the second container faces the first bottom side, The second opening communicates with the interior of the first container, A centrifuge tube comprising a plurality of the second containers.

4. The centrifuge tube according to claim 1 , wherein the first body portion and the cover member are fitted or threadedly engaged with each other.

5. 5. The centrifuge tube according to claim 1, wherein the lid member has a lid top surface portion covering the first opening of the first container, a lid opening is formed in the lid top surface portion, and the second container is positioned within the lid opening.

6. A kit for producing the centrifuge tube according to any one of claims 1 to 5, comprising: a first container having a first base, a first body, and a first opening; A cover material removably provided on the first opening of the first container; A kit comprising a second container having a second body and a second opening.

7. A process for preparing a centrifuge tube, comprising: The centrifuge tube comprises: a first container having a first base, a first body, and a first opening; A cover material removably provided on the first opening of the first container; a second container having a second body and a second opening; The width of the second opening of the second container is 0.2 μm or more and 2000 μm or less, The second container is held by the lid, The second opening of the second container faces the first bottom side, providing a centrifuge tube, the second opening being in communication with an interior of the first container; storing a first gelling solution in the first container and a second gelling solution in the second container; and a step of dripping droplets of the second gelling solution from the second container into the first gelling solution contained in the first container by applying centrifugal force to the centrifuge tube.

8. A step of preparing a centrifuge tube according to claim 3; accommodating a first gelling solution in any one of the second containers and a second gelling solution in the other second containers; and applying centrifugal force to the centrifuge tube to cause droplets of the first gelling solution to drip from a second container containing the first gelling solution into the first container, and cause droplets of the second gelling solution to drip from the second container containing the second gelling solution into the first container.

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