JP2022174649A - Centrifugal tube, kit and method for manufacturing particulate gel - Google Patents

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

The present disclosure relates to centrifuge tubes, kits, and methods of making particulate gels.

Conventionally, for example, as disclosed in Patent Literature 1, a technique is known in which a gelling solution is dripped by centrifugal force to produce a microparticle gel.

However, in the device described in Patent Document 1, the tubular portion for dripping is held within the gelation container. For this reason, when collecting the prepared microparticle gel, the tubular portion may hinder collection of the microparticle gel. In addition, it is necessary to carefully remove the tubular portion with sterilized tweezers or the like before collecting the microparticle gel, and there is a risk that complicated operations will be required to collect the microparticle gel without causing contamination. In addition, since the tubular portion is held in the gelation container, friction occurs at the portion where the tubular portion is held, which may generate foreign matter in the gelation container, which also causes contamination. can be a risk.

Further, for example, Patent Literature 2 discloses a technique of ejecting a gelling solution in the form of droplets by centrifugal force to produce a microparticle gel.

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, only the container containing the gelling solution is rotated when the centrifugal force is applied to the container. In this case, the gelling solution may scatter around, which may increase the size of the apparatus and increase the risk of contamination.

JP 2012-176374 A JP 2013-158753 A

The present disclosure provides a centrifuge tube, a kit, and a method for producing a microparticle gel that can suppress the occurrence of contamination when collecting the microparticle gel.

The centrifugal tube according to this embodiment includes a first container having a first bottom, a first body, and a first opening, and a lid detachably provided at the first opening of the first container. a second container having a material, 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; A container is held by the lid member, the second opening of the second container faces the first bottom side, and the second opening communicates with the inside of the first container.

In the centrifugal tube according to this embodiment, the first trunk and the lid member may be fitted or screwed together.

In the centrifuge tube according to the present embodiment, the lid member has a lid top surface portion that covers the first opening portion of the first container, and the lid top surface portion is formed with a lid opening. The second container may be located within the section.

In the centrifuge tube according to this embodiment, between the first container and the lid member, between the second container and the lid member, or between the first container and the second container A communicating portion may be provided that communicates the space to be filled with the outside air.

In the centrifugal tube according to the present embodiment, a liquid droplet guide section may be provided in the first container for guiding liquid droplets from the second opening of the second container.

In the centrifugal tube according to this embodiment, a plurality of the second containers may be provided.

The kit according to this embodiment is a kit for producing the centrifuge tube according to this embodiment, comprising a first container having a first bottom, a first body, and a first opening; A second container having a lid member detachably attached to the first opening of the first container, a second body, and a second opening is provided.

The method for producing a particulate gel according to this embodiment includes the steps of preparing the centrifuge tube according to this embodiment, storing a first gelling solution in the first container, and storing a second gelling solution in the second container. and applying a centrifugal force to the centrifugal tube to transfer the droplets of the second gelling solution from the second container to the first gelling solution contained in the first container and a step of dropping into.

The method for producing a microparticle 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 plurality of second containers, and , the step of accommodating the second gelling solution in another second container; and applying a centrifugal force to the centrifugal tube to remove the first gelling solution from the second container accommodating the first gelling solution. dropping droplets of a solution into the first container, and dropping droplets of the second gelling solution from a second container containing the second gelling solution into the first container. .

According to this embodiment, it is possible to suppress the occurrence of contamination when recovering the particulate gel.

FIG. 1 is a vertical cross-sectional view showing a centrifuge tube according to one embodiment. FIG. 2 is a vertical sectional view showing a centrifuge tube according to one variant. Figures 3(a)-(d) are diagrams illustrating the use of a centrifuge tube according to one embodiment. FIG. 4 is a vertical sectional view showing a centrifuge tube according to a first modification. FIG. 5 is a vertical sectional view showing a centrifuge tube according to a second variant. FIG. 6 is a vertical sectional view showing another configuration of the centrifugal tube according to the second modified example. 7 is an optical microscope photograph showing a microparticle gel produced using the centrifuge tube according to Example 1. FIG.

An embodiment will be specifically described below with reference to the drawings. Each figure shown below is shown typically. Therefore, the size and shape of each part are appropriately exaggerated for easy understanding. In addition, it is possible to modify and implement as appropriate without departing from the technical idea. In addition, in each figure shown below, the same code|symbol is attached|subjected to the same part and detailed description may be partially abbreviate|omitted. In addition, numerical values such as dimensions and material names of each member described in this specification are examples as an embodiment, and are not limited to these, and can be appropriately selected and used. In this specification, terms specifying shapes and geometrical conditions, such as parallel, orthogonal, and perpendicular terms, not only have strict meanings but also include substantially the same states. Also, for convenience of explanation, the terms "upper" and "lower" may be used, but the up-down direction may be reversed.

As used herein, "upward" refers to the central axis direction of the centrifuge tube and the direction facing away from the first bottom of the first container. Moreover, the term “downward” refers to the central axis direction of the centrifuge tube and the direction facing the first bottom side of the first container.

The present embodiment will be described below. This embodiment is not limited to what is described below.

[Centrifuge tube]
First, referring to FIG. 1, the configuration of the centrifugal tube according to this embodiment will be described. FIG. 1 is a cross-sectional view showing a centrifugal tube 10 according to this embodiment.

As shown in FIG. 1, the centrifugal tube 10 is a device for dropping a solution by centrifugal force to produce a fine particle gel with high size uniformity. This centrifuge tube 10 reduces the risk of contamination from the external environment and requires sterility, such as the preparation of microparticle gels as cell culture carriers for culturing cells on the surface and inside, and microparticle gels used in drug delivery systems. It is suitably used for the preparation of fine particles in the field where it is used. Moreover, it is particularly suitably used, for example, when liquid droplets of a sodium alginate solution are reacted with a calcium chloride solution to produce a microparticle gel of calcium alginate.

A centrifuge tube 10 according to this embodiment includes a first container 20 , a lid member 30 and a second container 40 . Among these, the first container 20 has a first bottom portion 21 , a first body portion 22 and a first opening portion 23 . The lid member 30 is detachably provided on the first opening 23 of the first container 20 . The second container 40 has a second body portion 41 and a second opening 42 , and the second container 40 is held by the lid member 30 . Further, 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.

[First container]
The first container 20 is, for example, a container for containing a solution such as a first gelling solution, and includes a first bottom portion 21 located below, a first trunk portion 22 connected to the first bottom portion 21, and a first trunk portion. and a first opening 23 located above the portion 22 . Among them, the first bottom portion 21 and the first body portion 22 form a space for containing the solution. The first opening 23 constitutes an opening for filling the first container 20 with the solution and taking out the solution from the first container 20 .

The configuration of the periphery of the first opening 23 in the first trunk portion 22 is not particularly limited as long as the lid member 30 can be attached and detached. For example, the first trunk portion 22 and the lid member 30 may be fitted to each other, or the first trunk portion 22 and the lid member 30 may be screw-engaged with each other. .

Also, the structure of the first bottom portion 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 such that the microparticle gel is accumulated at a predetermined position so that the microparticle gel formed in the first container 20 can be easily collected. In addition, the lowest point position where the particulate gel accumulates is not particularly limited, and may be the center position of the first bottom portion 21 (the position passing through the central axis CL), or may be shifted from the center position of the first bottom portion 21. It may be the edge. Further, it is more preferable that the lowest point position is shifted from directly below the second opening 42 of the second container 40 . The reason for this is as follows. The prepared microparticle gel accumulates upward from the lowest position, but when the microparticle gel reaches near the liquid surface of the first gelling solution, the microparticle gel becomes an obstacle to the first gelling solution. may be difficult to drop appropriately into the second gelling solution, making it difficult to produce a uniform microparticle gel. For this reason, the more the position of the lowest point is deviated from directly below the second opening 42, the less likely the microparticle gel will interfere with dropping, and the easier it is to produce a uniform microparticle gel.

In addition, the first body portion 22 is desirably made of a transparent material so that the amount of the solution can be easily seen from the outside. Moreover, it is more desirable that a scale is printed on the first body portion 22 .

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. Combined resin, C-FLEX, polystyrene, polyethylene terephthalate, glass, silicon resin, fluororesin, acrylic resin, urethane resin, etc. are preferably used. Also, the material of the first container 20 is desirably a hard material so as not to be deformed by the centrifugal force applied to the first container 20 .

[Lid material]
The lid member 30 is detachably provided at the first opening 23 of the first container 20 as described above. The lid member 30 has a lid top surface portion 31 that covers the first opening 23 of the first container 20 and a lid peripheral edge portion 32 that is provided on the peripheral edge of the lid top surface portion 31 and extends toward the first bottom portion 21 side. A lid opening 33 is formed in the lid top surface portion 31 , and the second container 40 is positioned in the lid opening 33 . The lid member 30 may have a portion that does not cover the first opening 23 of the first container 20, but it is preferable that the portion that does not cover is as small as possible. Moreover, the lid member 30 and the second container 40 held by the lid member 30 may partially exist inside the first container 20 , but at least a portion thereof may be located inside the first container 20 . It is preferably located above the portion 23 . This makes it possible to remove the lid member 30 and the second container 40 from the first container 20 by grasping the portion of the lid member 30 and the second container 40 positioned in the upward direction when recovering the microparticle gel. . Therefore, it is not necessary to carefully insert sterilized tweezers into the first container 20, and the microparticle gel can be collected with less risk of contamination.

The material of the lid member 30 is not particularly limited, and may be polyolefin resin such as polyethylene or polypropylene, polyvinyl chloride, ethylene-vinyl acetate copolymer resin, C-FLEX, polystyrene, polyethylene terephthalate, glass, silicone resin, fluorine. Resins, acrylic resins, urethane resins, and the like are preferably used. Also, 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 .

[Second container]
The second container 40 is a container that stores a solution such as a second gelling solution, for example, and has a second trunk portion 41 and a second opening portion 42 located below the second trunk portion 41 . Among them, the second body portion 41 is made of a substantially cylindrical member. The second opening 42 faces the first bottom 21 side, 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 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 positioned closer to the first bottom portion 21 of the first container 20 than the lid member 30 and faces the first bottom portion 21 side. In addition, when viewed from the second opening 42 side, it is sufficient that a part of the inside of the first container 20 can be confirmed. may be located on the opposite side of the The width W1 of the second opening 42 is narrowed, specifically, 0.2 μm or more and 2000 μm or less. Therefore, the solution contained in the second body portion 41 does not drop from the second opening portion 42 normally (when no force such as centrifugal force is applied). 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 toward the first container 20 side. The second opening 42 may be a hole formed in the bottom of the second container 40, and has a nozzle-like structure protruding downward from the bottom of the second container 40 so that the solution can be easily dripped. is more desirable.

A tapered portion 43 is preferably provided at the lower portion of the second body portion 41 . As a result, the solution such as the second gelling solution can be discharged from the second opening 42 with as little waste as possible. The shape and size of the second opening 42 can be selected as appropriate, so that the size of the droplet of the solution dropped from the second opening 42 can be adjusted. For example, it is desirable that the shape of the second opening 42 is 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 is, the easier it is to support the cells. Desirably, when the width W1 of the second opening 42 is larger than 10 μm, it is possible to prepare a microparticle gel larger than 10 μm, which is the approximate size of cells, and there is the advantage that cells can be more easily supported. Further, when microparticle gels supporting cells are suspended and cultured by, for example, swirl culture, the smaller the width W1 of the second opening 42, the smaller the microparticle gels produced are, which is advantageous in that they are easier to float. 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 the length of the longest portion on the plane. say. 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 . Further, 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 .

A plurality of second openings 42 may exist. By having a plurality of second openings 42, it is possible to increase the amount of droplets that can be dropped in a certain period of time, which is more desirable from the viewpoint of throughput. Also, in that case, if the interval between the second openings 42 is narrow, the dripping solution is likely to merge near the second openings 42, and as a result, the dripped 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 produce a uniform microparticle gel. Further, when the solutions dripped from the respective second openings 42 merge and gel together, the particulate gel adheres to the second openings 42 and prevents dripping. The longer the interval, the better. Specifically, the interval between the second openings 42 is preferably 1 or more times the width W1 of the second openings 42, more preferably 10 times or more, and 100 times or more. Spacing is even more preferred. Further, when three or more second openings 42 are provided, it is preferable that the intervals between all the second openings 42 and all the other second openings 42 are within the above range. Further, when the interval between the second openings 42 is smaller than 1 time the width W1 of the second openings 42, even if it is assumed that the dripping solution is always fused in the vicinity of the second openings 42 before being dripped. good. In this case, as described above, the sum of the widths W1 of the plurality of second openings 42 adjacent to each other at narrow intervals is preferably 0.2 μm or more and 2000 μm or less, preferably 1 μm or more and 1000 μm or less, and further 10 μm or more and 200 μm. It is more desirable to: This makes it difficult to drop the solution without centrifugation, as in the case where the dropped solution does not coalesce, and makes it easier to support the cells on the prepared microparticle gel and to facilitate rotation culture.

A means for holding the second container 40 on the lid member 30 is not limited. In other words, it is sufficient that the second container 40 does not come off from the lid member 30 when applying centrifugal force to the centrifuge tube 10 . In FIG. 1 , the second container 40 is penetrated by the lid opening 33 of the lid member 30 , and the upper portion 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. Further, the second container 40 and the lid member 30 may be of an integrated structure. Alternatively, a portion of the second container 40 may be sandwiched and fixed between the first container 20 and the lid member 30 at the connecting portion between the first container 20 and the lid member 30 . Furthermore, the second container 40 and the lid member 30 are not completely fixed, and the second container 40 and the lid member 30 are moved so that the position of the second container 40 can be adjusted vertically and horizontally in order to control the dropping conditions. It may be connected as possible. The position where the second container 40 is held is not particularly limited, and may be the central portion of the lid member 30 or the end portion of the lid member 30 .

The second container 40 further has a third opening 44 positioned above the second body 41 . The third opening 44 faces the opposite side of the second opening 42 and is an opening for adding a solution into the second container 40 . A flange portion 45 that holds the second container 40 to the lid member 30 is formed around the third opening portion 44 . In addition, in order to prevent contamination from the third opening 44, it is desirable to further provide a cover member (not shown) covering the third opening 44. FIG.

The material of the second container 40 is preferably transparent so that the amount of liquid in the second container 40 can be easily seen from the outside. Further, it is more desirable that a scale is printed on the second body portion 41 . Materials of the second container 40 include, for example, polyolefin resin such as polyethylene and polypropylene, polyvinyl chloride, ethylene-vinyl acetate copolymer resin, C-FLEX, polystyrene, polyethylene terephthalate, glass, silicon resin, fluororesin, acrylic resin, Urethane resin and the like are preferably used. Also, the material of the second container 40 is desirably a hard material so as not to be deformed by the centrifugal force applied to the second container 40 .

Further, as shown in FIG. 2, a plurality of second containers 40 may be provided. In this case, the plurality of second containers 40 may all contain the same type of solution, or the plurality of second containers 40 may contain different types of solutions. When the same type of solution is stored in a plurality of second containers 40, the amount of droplets that can be dropped in a certain period of time can be increased by simultaneously dropping the solutions from the plurality of second containers 40. possible and more desirable from a throughput standpoint. Also, in this case, the distance between the second opening 42 of the second container 40 and the second opening 42 of the second container 40 different therefrom is 1 times the width W1 of the second opening 42. The interval is preferably 10 times or more, more preferably 10 times or more, and even more preferably 100 times or more. By setting the interval between the second openings 42 within the above range, it is possible to suppress the tendency of the dripping solution to fuse near the second openings 42 and prevent the dripped solution from becoming larger than expected. . For this reason, the dripping of the solution can be suppressed without centrifugation, and it is possible to prevent difficulty in controlling dripping and difficulty in producing a uniform microparticle gel. In addition, when the solutions dripped from the respective second openings 42 are gelled by merging with each other, it is possible to suppress adhesion of fine particle gel to the second openings 42 and hindrance of dripping. When a plurality of second containers 40 contain different types of solutions, at least one of the second containers 40 contains the second gelling solution, and the other second containers 40 contain the cell suspension and the drug. Alternatively, the first gelling solution or the like may be accommodated.

[Entire centrifuge tube]
The centrifugal tube 10 preferably has excellent centrifugal strength. Excellent centrifugal strength means, for example, having strength to the extent that cracks do not occur in a centrifugal test at 1500 rpm. More preferably, the centrifugal tube 10 should not crack or split in the centrifugal test at 3000 rpm, more preferably in the centrifugal test at 4000 rpm. Examples of evaluation conditions for the centrifugation test include the following conditions. Centrifuge: HITACHI CF16RX, rotor used: swing rotor, adapter suitable for 50 mL centrifuge tube, set centrifugal strength: 1500 rpm or more, set temperature: 20°C, time: 10 minutes, content: pure water of 10 mL or more.

[Gelling solution]
The first gelling solution contained in the first container 20 and the first gelling solution contained in the second container 40 are not particularly limited as long as they mix with each other to form a gel. For example, as the first gelling solution, an aqueous solution of a high cation salt such as calcium chloride is preferably used, and as the second gelling solution, a sodium alginate solution is preferably used. When preparing a microparticle gel containing some substance such as cells or drugs, it is desirable to include the substance 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 surface tension such as a surface active substance or a substance capable of adjusting viscosity such as a thickener in order to control dripping. good.

[Action of the present embodiment]
Next, the operation of this embodiment having such a configuration will be described. Specifically, a method for producing a microparticle gel using the centrifugal tube 10 described above will be described.

First, as shown in FIG. 3(a), the aforementioned centrifuge tube 10 is prepared. The centrifugal tube 10 has each member (for example, the first container 20, the lid member 30, and the second container 40) configured separately, and can be assembled to have the structure of the centrifugal tube 10 when necessary. It may be a kit, in which case the centrifuge tube 10 is assembled before use. This embodiment also provides a kit for producing such a centrifuge tube.

Next, as shown in FIG. 3B, the first container 20 of the centrifuge tube 10 is filled with the first gelling solution L1, and the second container 40 is filled with the second gelling solution L2.

Subsequently, as shown in FIG. 3(c), a centrifugal force is applied to the centrifuge tube 10 (see the arrow in FIG. 3(c)). Due to this centrifugal force, the second gelling solution L2 is pushed out and passes through the second opening 42 having a narrow width W1. Thereby, droplets of the second gelling solution L2 can be dripped from the second opening 42 into the first gelling solution L1 accommodated in the first container 20 . The droplets of the second gelling solution L2 undergo a gelation reaction with the first gelling solution L1 in the first container 20 to generate particulate gel. In this case, the generated particulate gel can be accumulated on the first bottom portion 21 of the first container 20 .

When applying centrifugal force to the centrifuge tube 10, it is desirable to install the centrifuge tube 10 in a centrifuge (not shown) for centrifugation. The centrifuge rotor may be an angle rotor or a swing rotor. For example, it is desirable to use a swing rotor to apply centrifugal force directly downward to the centrifuge tube 10, and it is desirable to use an angle rotor to apply centrifugal force obliquely downward to the centrifuge tube 10. The direction in which the centrifugal force is applied can be controlled by adjusting the installation angle of the centrifugal tube 10 and the balance of the center of gravity position, in addition to selecting the rotor described above.

Next, as shown in FIG. 3D, after the dropping of the droplets of the second gelling solution L2 is completed, the second container 40 is removed from the first container 20 together with the lid member 30. Next, as shown in FIG. After that, using a pipette (not shown) or the like, the solution containing the microparticle gel is taken out from the first container 20 .

In this embodiment, the lid member 30 holds the second container 40 . Thereby, the second container 40 can be easily removed from the first container 20 by removing the lid member 30 after producing the microparticle gel. Moreover, the lid member 30 for attaching the second container 40 to the first container 20 does not exist inside the first container 20 . Therefore, it is possible to suppress the generation of foreign matter due to rubbing of the cover member 30 with the first container 20 in the first container 20, thereby reducing the risk of contamination. As a result, using a pipette or the like, the solution containing the microparticle gel can be easily recovered from the first container 20 with little risk of contamination. In addition, since the lid member 30 connects the second container 40 and the first container 20, the centrifugal tube 10 can be easily configured.

In addition, when a plurality of second containers 40 are provided as shown in FIG. Another second container 40 may contain the second gelling solution L2. In this case, by applying centrifugal force to the centrifugal 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. FIG. Further, droplets of the second gelling solution L2 are dropped into the first container 20 from the second container 40 containing the second gelling solution L2. 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 are fused in the first container 20 to prepare a particulate gel. may

As described above, according to the present 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 member 30. there is 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 fine particle gel having a highly uniform size is generated in the first container 20. be able to. Further, by removing the removable lid member 30 from the first container 20, the second container 40 held by the lid member 30 can also be easily removed. As a result, the particulate gel can be recovered from the first container 20 simply and with reduced risk of contamination.

[Modification]
Next, modifications of the present disclosure will be described with reference to FIGS. 4 to 6. FIG. 4-6 each illustrate a centrifuge tube according to a variation of the present disclosure. In FIGS. 4 to 6, the same reference numerals are assigned to the same parts as those in FIGS. 1 to 3, and detailed description thereof will be omitted.

(First modification)
FIG. 4 shows a vertical section through a centrifuge tube 10 according to a first variant. In FIG. 4 , a first communication portion 25 is formed between the first container 20 and the lid member 30 . A second communicating portion 34 is formed between the second container 40 and the lid member 30 . The first communication portion 25 and the second communication portion 34 communicate the space S existing between the first container 20 and the second container 40 with the outside air. Either one of the first communicating portion 25 and the second communicating portion 34 may be provided, or both of them may be provided.

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

The second communication portion 34 is a connecting portion between the second container 40 and the lid member 30 and is provided on the lid member 30 side. That is, the lid member 30 has a second communicating portion 34 . However, not limited to this, the second communication part 34 may be provided on the second container 40 side. That is, the second container 40 may have the second communicating portion 34 . Alternatively, the second communicating portion 34 may be a gap that is created between the second container 40 and the lid member 30 when the second container 40 and the lid member 30 are fitted together. The second communication part 34 may have a structure in which a gap is formed without completely blocking the connecting part between the second container 40 and the lid member 30 by a projection or the like. Alternatively, the second communication part 34 may be formed by providing a through-hole in the second container 40 and the lid member 30 at a position where there is no problem in containing the solution.

According to this modification, when 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 However, this positive pressure can be prevented from interfering with the dripping of the second gelling solution. Therefore, the first communicating portion 25 and the second communicating portion 34 are configured so that the gas inside and outside the space S communicates with each other and the positive pressure can be released.

The first communicating part 25 and/or the second communicating part 34 are not communicated when the second gelling solution is not dropped from the second container 40, and the second gelling solution is dropped by centrifugal load, and the space S is positive. It may have a valve structure that communicates when pressure is reached. Further, the structure may be such that the first communication portion 25 and/or the second communication portion 34 can be formed immediately before applying the centrifugal force. For example, when the lid member 30 and the first container 20 and/or the second container 40 are connected by screwing, the first communicating portion 25 and/or the second communicating portion 34 apply centrifugal force to the centrifugal tube 10. It may be formed by loosening the screw just prior to application. Alternatively, the first communicating portion 25 and/or the second communicating portion 34 may be closed with a cap (not shown), and the cap may be removed immediately before applying the centrifugal force to the centrifuge tube 10 to open the first communicating portion 25 and/or the second communicating portion . In this way, by closing the first communication portion 25 and/or the second communication portion 34 when unnecessary, foreign matter can pass through the first communication portion 25 and/or the second communication portion 34 The risk of intrusion into the container 20 can be reduced.

Also, a plurality of the first communication portion 25 and/or the second communication portion 34 may be provided. The cross-sectional area of the first communication part 25 and/or the second communication part 34 is preferably larger than the cross-sectional area of the second opening 42 of the second container 40 in order to facilitate the discharge of the positive pressure gas. desirable. Here, when a plurality of first communicating portions 25 and/or second communicating portions 34 are provided, the total cross-sectional area of all of them is the cross-sectional area.

In addition, in order to suppress sedimentation of foreign matter such as floating particles inside the first container 20 through the first communication portion 25 and/or the second communication portion 34, the first communication portion 25 and/or It is desirable that the exit of the second communicating portion 34 to the outside faces a direction other than the upward direction, such as the lateral direction or the downward direction of the centrifuge tube 10 . As a result, sedimentation of foreign matter inside the first container 20 through the first communicating portion 25 and/or the second communicating portion 34 can be suppressed. For example, in FIG. 4, the outlet of the first communicating portion 25 and the outlet of the second communicating portion 34 are directed in a direction other than the upward direction (lateral direction). Even if the outlet of the first communication portion 25 and/or the second communication portion 34 faces upward, the route of the first communication portion 25 and/or the second communication portion 34 is halfway. preferably curved. That is, if the cross section of any of the paths of the first communication portion 25 and/or the second communication portion 34 does not face upward, foreign matter may enter the first communication portion 25 and/or the second communication portion 34. can also be suppressed from settling inside the first container 20 .

(Second modification)
FIG. 5 shows a centrifuge tube 10 according to a second variant. In FIG. 5 , a liquid droplet guide section 50 is provided in the first container 20 to guide liquid droplets from the second opening 42 of the second container 40 . In this case, the liquid droplet guide section 50 may be an inclined plate that is inclined with respect to the central axis CL of the centrifugal tube 10 . At least a portion of the droplet guiding portion 50 is provided at a position closer to the first bottom portion 21 than the second opening 42 and spaced apart from the second opening 42 .

By the way, when the second opening 42 comes into contact with the first gelling solution in the first container 20, the second opening 42 may be clogged, preventing droplets from dripping. In addition, when the second opening 42 is completely immersed in the first gelling solution in the first container 20, the second gelling solution is discharged from the second opening 42 in the form of a string rather than droplets. It may interfere with proper dripping. Therefore, 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 when the droplet dropped from the second opening 42 falls into the first gelling solution becomes large. , the droplet may be deformed or collapsed.

On the other hand, in this modified example, a liquid droplet guide section 50 is provided in the first container 20 to guide liquid droplets from the second opening 42 of the second container 40 . The droplet guide section 50 is arranged obliquely with respect to the direction of centrifugal force (the direction of the arrow in FIG. 5). As a result, the droplet dropped from the second opening 42 is allowed to fall into the first gelling solution via the droplet guiding portion 50, and the impact of the droplet dropping into the first gelling solution can be softened. . Also, the risk of the second opening 42 of the second container 40 coming into contact with the first gelling solution can be reduced.

Note that the position and orientation of the liquid droplet guide section 50 are not limited to this. Also, the liquid droplet guide section 50 may or may not be fixed to the first container 20 . It is preferable that the liquid droplet guide section 50 has a structure such that liquid droplets move diagonally with respect to the direction of the centrifugal force when the centrifugal force is applied. Further, in order to soften the impact when the droplets dropped from the second opening 42 fall on the droplet guiding portion 50, it is desirable that the distance between the droplet guiding portion 50 and the second opening 42 is short. More preferably, the droplet guiding portion 50 and the second opening 42 are in contact with each other.

The cross-sectional shape in the width direction of the liquid droplet guide section 50 (the cross-sectional shape cut along a plane P perpendicular to the length direction of the liquid droplet guide section 50 (the direction of arrow A in FIG. 5)) is such that the liquid droplet It is desirable to have a shape that can be induced into a gelling solution. For example, it is desirable that the liquid droplet guiding portion 50 has a slide-like structure in which the central portion in the width direction is lower than the end portions in the width direction. Specifically, the cross-sectional shape in the width direction of the liquid droplet guiding portion 50 is preferably U-shaped or V-shaped, for example. As a result, the droplets pass through the width direction center of the droplet guiding portion 50 and fall into the first gelling solution, which prevents the droplets from jumping out of the droplet guiding portion 50, thereby improving the fine particle gel production. Reproducibility can be improved. In addition, the fact that the central portion in the width direction of the liquid droplet guide portion 50 is lower than the end portions in the width direction means that a plurality of the second containers 40 are provided, and the liquid droplets dropped from the respective second containers 40 are merged. Also desirable (see FIG. 2). In this case, since the droplets gather at the central portion in the width direction of the droplet guiding portion 50, the droplets can be easily fused.

It is desirable that the surface of the droplet guiding portion 50 has high hydrophobicity so that droplets can easily slide. The contact angle of the surface of the droplet guiding portion 50 to water is desirably 60 degrees or more, more desirably 90 degrees or more, and even more desirably 120 degrees or more.

The material of the droplet guiding part 50 is the same material as the first container 20, or a material different from 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. It is desirable that the material is also soft. In addition, in order to prevent the droplet guiding part 50 from being an obstacle when collecting the particulate gel in the first container 20, the particulate gel can be collected with a pipette or the like without removing the droplet guiding part 50. It is desirable that Specifically, it is desirable that the shape and arrangement of the droplet guiding portion 50 be such that the tip of the pipette can be brought close to the first bottom portion 21 of the first container 20 while avoiding the droplet guiding portion 50. .

As shown in FIG. 6 , the liquid droplet guiding portion 50 may be configured from the inner surface of the first body portion 22 of the first container 20 . In this case, by applying a centrifugal force obliquely downward (in the direction of the arrow in FIG. 6) with respect to the central axis CL of the centrifugal tube 10, the droplet dropped from the second opening 42 is first moved to the first container 20. is dropped on the inner surface of the first trunk portion 22 of the. A droplet can then fall through the inner surface of the first body 22 into the first gelling solution. 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 arranged near some portion of the inner surface of the first barrel 22 . Specifically, the second container 40 may be held at a position offset from the center of the lid member 30, the second container 40 may be held at the edge of the lid member 30, or the second container 40 may be held by the lid member 30. You may hold|maintain so that it may become diagonal with respect to. Thereby, the second opening 42 may be brought closer to the inner surface of the first body 22 . Further, when a centrifugal force is applied obliquely downward with respect to the central axis CL of the centrifuge tube 10, when the centrifugal force is removed, gravity causes the flow of the first gelling solution in the direction opposite to the direction of the centrifugal force, The impact may cause the first gelling solution to touch the second opening 42 . In order to reduce such risk, the position of the second opening 42 is deviated from the central axis CL of the centrifuge tube 10 and arranged near any portion of the inner surface of the first barrel 22. is desirable.

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

(Example 1)
A centrifuge tube shown in FIG. 1 was prepared. In this case, first, a 50 ml conical tube manufactured by FALCON (product number 352070) was prepared as the first container and lid. Also, a 2.5 ml syringe manufactured by Terumo Corporation (product number SS-02LZ) was prepared as a second container. Next, a through hole was made in the center of the cover member, and the second container was passed through the through hole, and attached so that the flange portion of the second container was caught and held by the cover member. At this time, it was confirmed that even if the plunger of the second container was pushed to send air into the first container, the positive pressure inside the first container was immediately released. A metal needle manufactured by Musashi Engineering (32G, product number SNA-32G-B) 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 410 G was applied directly below the centrifuge tube for 15 minutes. This resulted in almost all of the sodium alginate solution being added dropwise. As measured by optical microscope observation, a fine particle gel with a diameter of about 200 μm and a high size uniformity was formed on the first bottom of the first container (see FIG. 7). After that, the lid member and the second container were removed with bare hands, and the microparticle gel could be collected with a pipette. In addition, as a lid material, a lid material having a through hole provided in the center in advance without opening a through hole was produced by a 3D printer, and the same test as described above was performed, and similar results were obtained.

(Example 2)
A centrifugal tube having a droplet guiding portion as shown in FIG. 5 was produced. In this case, first, a centrifuge tube was produced in substantially the same manner as in Example 1. On the other hand, as a liquid droplet guiding portion, a polypropylene rectangular plate of 65 mm long×5 mm wide was prepared, and this plate was placed diagonally in the first container. At this time, the distance between the second opening and the droplet guiding portion was approximately 5 mm. Next, 5 ml of 1 w % aqueous calcium chloride 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. In this state, as in Example 1, a centrifuge was used to apply a centrifugal force of 410 G directly below the centrifuge tube for 15 minutes. As a result, almost all of the sodium alginate solution was dropped, and fine particle gel with a diameter of about 200 μm and high uniformity in size was formed on the first bottom of the first container. At this time, no fibrous substance was observed to disperse, as if the fine particle gel had collapsed. Furthermore, almost all of the solution in the first container could be recovered with a pipette without removing the droplet guiding part when recovering the fine particle gel.

It is also possible to appropriately combine a plurality of constituent elements disclosed in the above embodiments and modifications as necessary. Alternatively, some components may be deleted from all the components shown in the above embodiments and modifications.

REFERENCE SIGNS LIST 10 centrifugal tube 20 first container 21 first bottom 22 first body 23 first opening 24 inclined portion 25 first communicating portion 30 lid member 31 lid top surface portion 32 lid peripheral edge portion 33 lid opening portion 34 second communicating portion 40 second container 41 second barrel 42 second opening 43 tapered portion 44 third opening 45 flange 50 droplet guide

Claims (9)

a first container having a first bottom, a first body, and a first opening;
a cover member removably provided at 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 member,
the second opening of the second container faces the first bottom side,
The centrifugal tube, wherein the second opening communicates with the inside of the first container. 2. The centrifugal tube according to claim 1, wherein said first barrel and said lid member are fitted or screw-engaged with each other. The lid member has a lid top surface that covers the first opening of the first container, the lid top surface has a lid opening, and the second container is positioned in the lid opening. 3. The centrifuge tube of claim 1 or 2, comprising: Between the first container and the lid member or between the second container and the lid member, the space existing between the first container and the second container is communicated with the outside air. 4. The centrifugal tube according to any one of claims 1 to 3, wherein a communicating portion is provided. 5. The centrifuge tube according to any one of claims 1 to 4, wherein the first container is provided with a droplet guide section that guides droplets from the second opening of the second container. 6. The centrifuge tube according to any one of claims 1 to 5, wherein a plurality of said second containers are provided. A kit for making the centrifuge tube according to any one of claims 1 to 5,
a first container having a first bottom, a first body, and a first opening;
a cover member detachably provided at the first opening of the first container;
A kit comprising a second body and a second container having a second opening. providing a centrifuge tube according to any one of claims 1 to 5;
housing a first gelling solution in the first container and housing a second gelling solution in the second container;
applying a centrifugal force to the centrifugal tube to drop droplets of the second gelling solution from the second container into the first gelling solution contained in the first container; A method for producing a particulate gel, comprising: providing a centrifuge tube according to claim 6;
a step of accommodating the first gelling solution in one of the plurality of second containers and accommodating the second gelling solution in the other second containers;
By applying centrifugal force to the centrifugal tube, droplets of the first gelling solution are dropped from the second container containing the first gelling solution into the first container, and the second gelling dropping droplets of the second gelling solution from a second container containing the solution into the first container.

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