JP7271974B2 - Separation vessel kit - Google Patents

Description

The present invention relates to a separate container kit.

Centrifugation using a density gradient, for example, is widely used to separate sperm from semen, blood cells from blood, and the like. The density gradient centrifugation method can be performed, for example, as follows. First, a separation liquid such as a Percoll solution is placed in a centrifugation tube, a semen sample is layered on the Percoll liquid, and centrifuged. This creates a density gradient and the sperm sediment and separate into the bottom layer. Therefore, the sperm can be recovered by inserting a pipette into the centrifuge tube, reaching the tip of the pipette to the bottom layer, and aspirating. However, since the pipette is inserted from the top of the centrifuge tube and the tip is moved to the bottom layer, the components existing near the middle layer of the Percoll solution adhere to the pipette. Therefore, there is a problem that unnecessary components are mixed in the collected sperm sample. Therefore, the development of a separation container to avoid such problems is underway (Patent Document 1).

JP 2009-119457 A

The present inventor has developed a new separation container composed of an outer container and an inner container as a separation container for preventing such contamination of unnecessary components. The separation container has a cylindrical inner container with open top and bottom ends, and a bottomed cylindrical outer container with an open top end and a closed bottom end. Further, in a state in which the inner container is inserted into the outer container, the space between the outer container and the inner container serves as a collection part for the sperm, which is the object to be separated. For this separation container, for example, first, the inner container is inserted into the outer container, and the Percoll solution is introduced into the inner container. Then, a semen sample is overlaid on the Percoll solution and centrifuged. As a result, the spermatozoa in the semen layered on the inner container are sedimented, and the spermatozoa are introduced into the outer container through the opening at the lower end of the inner container. After centrifugation, a cap is attached to the upper end opening of the inner container to make the inside of the inner container liquid-tight. You can remove the device. Since the sedimented spermatozoa are present in the outer container, the spermatozoa can be recovered without inserting a pipette into the Percoll solution containing impurities, as described above.

However, when the separation container having the outer container and the inner container is used, the residual amount of the supernatant when the inner container is removed increases from the volume of the recovery unit, resulting in a decrease in sperm concentration. occured. Moreover, in reproductive medicine, it is believed that a higher sperm concentration at the time of collection is better from the viewpoint of fertilization rate or cryopreservation.

Accordingly, the first object of the present invention is to provide a separation container kit capable of suppressing an increase in the remaining amount of supernatant after removal of the inner container.

In addition, in a separation container that is used with the outer container and the inner container in contact, if the outer container and the inner container are stored in contact, the inner container will conform to the outer container, that is, the inner circumference of the outer container The outer peripheral shape of the inner container is deformed according to the shape. In addition, if the outer container and the inner container are stored in contact with each other, stress is applied to the inner container and the outer container during storage, which may cause damage such as cracks during use.

Therefore, it is a second object of the present invention to provide a separation container kit capable of reducing the fitting of the inner container to the outer container or the occurrence of damage such as cracks during use.

In order to achieve the first object, the separation container kit (hereinafter also referred to as "separation kit") for separating an object to be separated from a liquid sample of the present invention includes a separation container and a cap,
The separation container is
having an outer container, an inner container, and a first pressure contact part,
The outer container is
It has a bottomed cylindrical shape with an open upper end and a closed lower end,
The inner container is
It has a cylindrical shape with an upper end and a lower end opened,
The inner container and the outer container are
The inner container can be inserted into the outer container in a state in which the respective vertical directions are aligned,
In a state in which the inner container is inserted into the outer container, the space between the outer container and the inner container is a separation target object recovery unit,
The cap is
Having a fitting portion for the upper end opening of the inner container,
The fitting portion can make the inside of the inner container liquid-tight by fitting into the upper end opening of the inner container,
In a state in which the inner container is inserted into the outer container, the inner peripheral surface of the outer container and the outer peripheral surface of the inner container are pressed together by the first pressure contact portion,
It is characterized in that the recovery portion is brought into a liquid-tight state by the pressure contact.

In order to achieve the second object, a separation container kit for separating an object to be separated from a liquid sample of the present invention comprises a separation container and a cap,
The separation container is
having an outer container, an inner container, and a temporary fixing portion,
The outer container is
It has a bottomed cylindrical shape with an open upper end and a closed lower end,
The inner container is
It has a cylindrical shape with an upper end and a lower end opened,
The inner container and the outer container are
The inner container can be inserted into the outer container in a state in which the respective vertical directions are aligned,
In a state in which the inner container is inserted into the outer container, the inner peripheral surface of the outer container and the outer peripheral surface of the inner container come into contact with each other in the circumferential direction, so that the outer container and the inner container are separated from each other. A space is formed between, and the space is a recovery part for the separation target,
The cap is
Having a fitting portion for the upper end opening of the inner container,
The fitting portion can make the inside of the inner container liquid-tight by fitting into the upper end opening of the inner container,
The temporary fixing portion temporarily fixes the inner container to the outer container, and in a state in which the inner container is temporarily fixed to the inside of the outer container, the inner peripheral surface of the outer container and the outer peripheral surface of the inner container. It is characterized in that contact in the circumferential direction of is inhibited.

ADVANTAGE OF THE INVENTION According to this invention, the increase in the residual amount of the supernatant after removal of the said inner container can be suppressed.

Moreover, according to the present invention, it is possible to reduce the conformability of the inner container to the outer container or the occurrence of damage such as cracks during use.

FIG. 1 is a cross-sectional view showing an example of the separation container in the separation container kit of the present invention, (A) is the outer container, (B) is the inner container, and (C) is the outer container. FIG. 4 is a cross-sectional view showing a separation container with an inner container inserted therein; FIG. 2 is a schematic diagram showing an example of the cap in the separation container kit of the present invention, (A) is a side view, and (B) is a cross-sectional view in the AA direction in FIG. 2 (A). (C) is a perspective view from below. FIG. 3 is a cross-sectional view showing the cap attached to the separation container in the separation container kit of the present invention. FIG. 4 is a cross-sectional view showing another example of the separation container in the separation container kit of the present invention, (A) is the outer container, (B) is the inner container, and (C) is the outer container. FIG. 4 is a cross-sectional view showing a separation container with an inner container inserted inside the container; FIG. 5 is a view showing another example of the separation container kit of the present invention, which is a partially enlarged cross-sectional view around the tip of the outer container. FIG. 6 is a cross-sectional view showing another example of the separation container in the separation container kit of the present invention, (A) is a partially enlarged cross-sectional view around the tip of the outer container, (B) and (C). [Fig. 2] is a partially enlarged cross-sectional view of the periphery of the upper end of the outer container; FIG. 7 shows another example of the separation container kit of the present invention, which is a partially enlarged cross-sectional view around the upper end of the outer container. FIG. 8 is a schematic diagram showing another example of the separation container of the separation container kit of the present invention, (A) is a partially enlarged side view around the opening of the outer container, and (B) is the inner container. is a partially enlarged cross-sectional view of the cover portion of the.

<Separation container kit>
The separation container kit for separating an object to be separated from a liquid sample of the present invention includes a separation container and a cap as described above,
The separation container is
having an outer container, an inner container, and a first pressure contact part,
The outer container is
It has a bottomed cylindrical shape with an open upper end and a closed lower end,
The inner container is
It has a cylindrical shape with an upper end and a lower end opened,
The inner container and the outer container are
The inner container can be inserted into the outer container in a state in which the respective vertical directions are aligned,
In a state in which the inner container is inserted into the outer container, the space between the outer container and the inner container is a separation target object recovery unit,
The cap is
Having a fitting portion for the upper end opening of the inner container,
The fitting portion can make the inside of the inner container liquid-tight by fitting into the upper end opening of the inner container,
In a state in which the inner container is inserted into the outer container, the inner peripheral surface of the outer container and the outer peripheral surface of the inner container are pressed together by the first pressure contact portion,
It is characterized in that the recovery portion is brought into a liquid-tight state by the pressure contact.

As a result of intensive research by the present inventor, it was found that the increase in the remaining amount of the supernatant that occurs when the inner container is removed is due to the following causes. That is, in the separation container, when a cap is attached to the upper end opening of the inner container, the insides of the outer container and the inner container can be made liquid-tight. Movement of the separation liquid is possible within the outer container and the inner container. When introducing the separated liquid into the inner container, the cap is not attached to the inner container. Therefore, when the separated liquid is introduced into the inner container, the separated liquid enters between the inner peripheral surface of the outer container and the outer peripheral surface of the inner container in addition to the collecting portion. Therefore, it was found that the separated liquid that permeated when the inner container was pulled out remained as the supernatant, and as a result, the residual amount of the supernatant increased from the volume of the recovery section. The separation container kit is provided with a first pressure contact portion, and in a state in which the inner container is inserted into the outer container, the inner peripheral surface of the outer container and the outer peripheral surface of the inner container are pressed against each other by the first pressure contact portion. The present inventors have found that an increase in the remaining amount of the supernatant can be suppressed by making the collection portion liquid-tight by pressing the collection portion with the pressure contact portion, and have completed the present invention. Therefore, according to the separation container kit of the present invention, it is possible to suppress an increase in the remaining amount of the supernatant after the removal of the inner container. Therefore, according to the separation container kit of the present invention, for example, the concentration of the separation target after separation can be increased. Further, in the separation container kit of the present invention, for example, after the separation, the collecting section is pressurized to have a positive pressure. Therefore, according to the separation container kit of the present invention, for example, when the inner container is removed, the recovery unit is unlikely to become negative pressure, so the negative pressure causes the transfer of the supernatant from the inner container to the recovery unit (liquid who) can be suppressed. Therefore, according to the separation container kit of the present invention, it is possible to suppress an increase in the remaining amount of the supernatant after the removal of the inner container.

In the present invention, the "axial direction" means the axial direction of the inner container, the outer container and the cap, and means the vertical direction, and the "vertical direction" means the direction perpendicular to the axial direction. , "height" refers to the length in the axial direction, and "diameter" refers to the diameter in the vertical cross section.

In the present invention, "pressure contact" means, for example, a state in which two members are in direct or indirect contact, and pressure is generated between the two members from one to the other and from the other to the other. do.

In the present invention, "fitting" includes, for example, fitting by inserting (inserting) and fitting by screwing (screwed), and may have any meaning.

In the separation container kit of the present invention, the separation container has the outer container and the inner container. The outer container and the inner container may each have a cylindrical shape, but a cylindrical shape is particularly preferable.

The shape and the like of the outer container and the inner container are not particularly limited as long as the inner container can be inserted into the outer container.

It is preferable that the inner container and the outer container each have a corresponding locking portion. With such a configuration, for example, when the inner container is inserted into the outer container, the outer container and the inner container can be locked by bringing the locking portions into contact with each other. . Moreover, when the inner container is inserted into the outer container, the position of the inner container in the outer container can be kept constant. When the engaging portion is formed on the inner peripheral surface of the outer container, the engaging portion of the inner container is formed on the outer peripheral surface of the inner container. Further, when the locking portion is formed on the outer peripheral surface of the outer container, the inner container has a cover portion that covers a part of the outer peripheral portion of the outer container, and the locking portion of the inner container comprises: It is formed on the inner peripheral surface of the cover portion. The locking portion may be, for example, a fitting portion. In this case, since the locking portion also serves as a moving means, which will be described later, for example, it is preferable to use a fitting portion by screwing, that is, a screwing portion.

The first pressure contact portion presses an inner peripheral surface of the outer container and an outer peripheral surface of the inner container in a state in which the inner container is inserted into the outer container, and the pressure contact causes the recovery portion to move. Make it liquid-tight. The first pressure-contact portion, for example, directly or indirectly presses the inner peripheral surface of the outer container and the outer peripheral surface of the inner container. The first pressure contact portion may be a separate member arranged between the inner peripheral surface of the outer container and the outer peripheral surface of the inner container, or may be a separate member arranged between the inner peripheral surface of the outer container and the outer peripheral surface of the inner container. It may be formed as a part. Examples of the separate member include a sealing member such as an O-ring. The sealing member preferably has elasticity. By adopting the sealing member, the separation container kit of the present invention can achieve wider dimensional accuracy and design for the outer container and the inner container compared to, for example, the case of adopting a pair of tapered portions described later. Even with a tolerance of , pressure contact between the inner peripheral surface of the outer container and the outer peripheral surface of the inner container can be achieved. When the first pressure contact portion is part of the outer container and the inner container, the first pressure contact portion may be, for example, a pair of corresponding tapered portions of the outer container and the inner container.

The position of the first pressure contact portion in the outer container and the inner container is, for example, the lower end of the inner container, the upper end of the inner container, the end surface of the opening of the inner container, or a position facing the outer container. , a portion adjacent to the collecting portion, and the like. The position of the first pressure contact portion is preferably adjacent to the recovery portion. As a result, the separation container kit of the present invention can more reliably prevent the separation liquid from permeating between the inner peripheral surface of the outer container and the outer peripheral surface of the inner container, for example.

When the first pressure contact portion is a pair of corresponding tapered portions of the outer container and the inner container, the outer container has, for example, the first inner peripheral tapered portion on the inner peripheral surface, and the inner container has a first outer peripheral taper on its outer peripheral surface. In this case, the inner peripheral surface of the first inner peripheral tapered portion and the outer peripheral surface of the first outer peripheral tapered portion are in direct contact with each other, so that the inner peripheral surface of the outer container and the outer peripheral surface of the inner container are pressed against each other. By the pressure contact, the collecting portion becomes liquid-tight. Thus, in the separation container kit of the present invention, the outer container and the inner container are brought into contact with each other by a taper, that is, they are brought into pressure contact with each other, so that the liquid-tight state of the recovery section can be achieved more reliably. Further, in the separation container kit of the present invention, for example, by changing the shape of the outer container and the inner container, the inner peripheral surface of the outer container and the outer peripheral surface of the inner container can be press-contacted, and a separate member is used for the press-contact. Since it is not necessary, the manufacturing cost can be reduced and it is excellent in terms of cost. For the outer container, for example, as the first inner peripheral tapered portion, only the inner peripheral surface may be tapered, or both the inner peripheral surface and the outer peripheral surface may be tapered. Further, the inner container may be formed with a taper only on the outer peripheral surface as the first outer peripheral tapered portion, or may be formed with a taper on the inner peripheral surface and the outer peripheral surface. The first inner peripheral tapered portion and the first outer peripheral tapered portion have, for example, a reverse tapered shape in which the inner diameter narrows from the top opening of the outer container toward the bottom surface. Moreover, it is preferable that the taper angle of the first inner peripheral taper portion and the taper angle of the first outer peripheral taper portion are substantially the same. Accordingly, in the separation container kit of the present invention, the outer container and the inner container can be brought into pressure contact with each other over a wider surface, so that, for example, the liquid-tight state of the recovery section can be more reliably achieved. When the first press-contact portion is a tapered portion, the separation container kit of the present invention is preferably configured in combination with, for example, moving means described later. Thus, in the separation container kit of the present invention, by applying an axial force to the inner container by the moving means, the first inner peripheral tapered portion and the first outer peripheral tapered portion are pressed against each other. Since a larger pressure can be applied to the surface, the liquid-tight state of the recovery section can be achieved more reliably.

The ratio (I:O) between the inner diameter (I) of the first inner peripheral taper portion and the outer diameter (O) of the first outer peripheral taper portion is, for example, 1:1.0003 or more, or 1:1. 0.0047 or greater. Said I:O ratio is preferably 1:1.0003 to 1:1.003, 1:1.0047 to 1:1.023, 1:1.0047 to 1:1.017, 1:1. 0047 to 1:1.005. Thereby, in the separation container kit of the present invention, the inner peripheral surface of the first inner peripheral taper portion and the outer peripheral surface of the first outer peripheral taper portion are formed in a state where the inner container is inserted into the outer container. The liquid-tight state of the recovery section can be more reliably achieved by the pressure contact with the . The inner diameter of the first inner peripheral tapered portion means the diameter of the inner peripheral surface of the outer container. Also, the outer diameter of the first outer peripheral tapered portion means the diameter of the outer peripheral surface of the inner container. The outer diameter and inner diameter are the outer diameter and inner diameter when the inner container is not inserted into the outer container. The I:O ratio is, for example, the ratio of corresponding portions of the first inner tapered portion and the first outer tapered portion. The corresponding portion is, for example, a portion where the inner container is inserted into the outer container and the inner peripheral surface of the first inner peripheral taper portion and the outer peripheral surface of the first outer peripheral taper portion are in pressure contact. , the inner circumference of the first inner circumference taper and the outer circumference of the first outer circumference taper having the same height. When the height of the first inner peripheral taper portion and the height of the first outer peripheral taper portion are the same, the state where the upper end and the lower end of the first inner peripheral taper portion and the first outer peripheral taper portion are aligned 3, the inner diameter of the first inner peripheral tapered portion and the outer diameter of the first outer peripheral tapered portion at the same height preferably satisfy the above ratio. When the height of the first inner circumference taper portion and the height of the first outer circumference taper portion are different, the inner diameter of the lower end of the first inner circumference taper portion and the lower end of the first outer circumference taper portion It is preferable that the outer diameter satisfies the above ratio.

The height (L _T1 ) of the first inner peripheral tapered portion is not particularly limited. The lower limit of _LT1 is, for example, 1 mm or more, preferably 5 mm or more, and more preferably 8 mm or more. The range of L _T1 is, for example, in the range of 1-5 mm, preferably in the range of 3-5 mm.

The height (L _T2 ) of the first outer peripheral tapered portion is not particularly limited. The lower limit of _LT2 is, for example, 1 mm or more, preferably 3 mm or more, and more preferably 5 mm or more. The range of _LT2 is, for example, in the range of 1-5 mm, preferably in the range of 3-5 mm.

The separation container kit of the present invention may have, for example, a plurality of pressure contacts. In this case, the separation container kit of the present invention has, for example, a second pressure contact portion, and in a state in which the inner container is inserted into the outer container, the inner peripheral surface of the outer container and the inner container are separated from each other. When the outer peripheral surface is in pressure contact with the first pressure contact portion, the inner peripheral surface of the outer container and the outer peripheral surface of the inner container are not in pressure contact with the second pressure contact portion. Further, when the inner container moves toward the bottom surface of the outer container and the pressure contact between the inner peripheral surface of the outer container and the outer peripheral surface of the inner container by the first pressure contact portion is released, the outer container It is preferable that the inner peripheral surface and the outer peripheral surface of the inner container are brought into pressure contact with each other by, for example, the second pressure contact portion, and the recovery portion is brought into a liquid-tight state by the pressure contact of the second pressure contact portion. Thus, in the separation container kit of the present invention, for example, even if the pressure contact by the first pressure contact portion is released by moving the inner container too far toward the bottom surface of the outer container, the second The pressure contact portion can make the collection portion liquid-tight. Therefore, according to the separation container kit of the present invention, for example, since the liquid-tight state of the recovery section can be more reliably achieved, an increase in the remaining amount of the supernatant after removal of the inner container can be more reliably suppressed. . For the specific example of the second pressure contact portion and the description of its position, for example, the description of the first pressure contact portion can be used.

It is preferable that the second pressure contact portions are a pair of corresponding tapered portions of the outer container and the inner container. In this case, the outer container has, for example, a second inner peripheral tapered portion on the inner peripheral surface, and the inner container has a second outer peripheral tapered portion on the outer peripheral surface. In this case, the inner peripheral surface of the second inner peripheral tapered portion and the outer peripheral surface of the second outer peripheral tapered portion are in direct contact with each other, so that the inner peripheral surface of the outer container and the outer peripheral surface of the inner container are pressed against each other. By the pressure contact, the collecting portion becomes liquid-tight. Thus, in the separation container kit of the present invention, the outer container and the inner container are brought into contact with each other by a taper, that is, they are brought into pressure contact with each other, so that the liquid-tight state of the recovery section can be achieved more reliably. Further, in the separation container kit of the present invention, for example, by changing the shape of the outer container and the inner container, the inner peripheral surface of the outer container and the outer peripheral surface of the inner container can be press-contacted, and a separate member is used for the press-contact. Since it is not necessary, the manufacturing cost can be reduced and it is excellent in terms of cost. For the outer container, for example, as the second inner peripheral tapered portion, only the inner peripheral surface may be tapered, or both the inner peripheral surface and the outer peripheral surface may be tapered. Further, the inner container may have, for example, a taper formed only on the outer peripheral surface as the second outer peripheral tapered portion, or may have a taper formed on the inner peripheral surface and the outer peripheral surface.

When the separation container kit of the present invention has the first pressure contact portion and the second pressure contact portion, the first pressure contact portion is formed adjacent to the recovery portion, and the second pressure contact portion is formed, for example. Preferably, the portion is formed on at least one of the inner peripheral surface of the upper end of the inner container and the outer peripheral surface of the outer container facing the inner peripheral surface of the upper end of the inner container. Thus, in the separation container kit of the present invention, by inserting the inner container into the outer container, the recovery unit can be made liquid-tight by the first pressure contact portion. Further, according to the separation container kit of the present invention, for example, even if the pressure contact by the first pressure contact part is released by moving the inner container too much toward the bottom surface of the outer container, the second 2, the recovery section can be liquid-tight. Therefore, according to the separation container kit of the present invention, for example, it is possible to more reliably suppress an increase in the remaining amount of the supernatant after the removal of the inner container.

The inner container may, for example, have a tip portion at its lower end. In this case, the tip portion is cylindrical, and the outer peripheral surface thereof does not contact the inner peripheral surface of the outer container when the inner container is inserted into the outer container. For example, the tip portion can also be referred to as a protruding portion that protrudes from the inner container toward the collecting portion when the inner container is inserted into the outer container. When the tip portion is projected toward the collecting portion and a space is provided between the outer peripheral surface of the tip portion and the outer container, the lower end of the tip portion is positioned, for example, in the supernatant. As a result, the separation container kit of the present invention can, for example, suppress the inflow of air into the inner container when the inner container is removed. Therefore, the separation container kit of the present invention can suppress the movement of the supernatant from the inner container to the collection unit caused by the inflow of air into the inner container, for example, so that the upper side of the inner container after removal of the inner container can be suppressed. It is possible to more reliably suppress an increase in the remaining amount of clean liquid.

The separation container of the present invention preferably has, for example, moving means capable of moving the inner container toward the bottom surface of the outer container, and the inner container is inserted into the outer container by the moving means. . The moving means, for example, is operated by the user of the separation container kit of the present invention to move the inner container toward the bottom surface of the outer container. Accordingly, in the separation container kit of the present invention, for example, the inner container can be easily inserted into the outer container, so that the recovery section can be easily brought into a liquid-tight state. It is preferable that the moving means can adjust, for example, the moving distance of the inner container in the axial direction.

The moving means includes, for example, corresponding threaded portions of the outer container and the inner container. As a specific example, the outer container may have a first threaded portion for the inner container, and the inner container may have a second threaded portion for the first threaded portion. . In this case, when the first screwing portion and the second screwing portion are screwed together and the inner container is inserted into the outer container, the inner peripheral surface of the outer container and the inner container is brought into pressure contact with the first pressure contact portion, and the recovery portion becomes liquid-tight due to the pressure contact. Thus, in the separation container kit of the present invention, for example, since the axial moving distance of the inner container can be adjusted, by adjusting the moving distance, the inner peripheral surface of the outer container can be moved by the first pressure contact portion. and the outer peripheral surface of the inner container. Therefore, in the separation container kit of the present invention, for example, by gradually opening the liquid-tight state of the recovery section, it is possible to suppress disturbance of the separated layer interface due to rapid withdrawal of the inner container. Further, when the inner container is suddenly removed from the outer container, for example, the collecting section becomes negative pressure, and the supernatant moves from the inner container to the collecting section. The separation container kit of the present invention can prevent the inner container from being abruptly removed from the outer container when the inner container is removed. movement of the supernatant from to the collection unit can be suppressed. Therefore, according to the separation container kit of the present invention, for example, it is possible to more reliably suppress an increase in the remaining amount of the supernatant after the removal of the inner container.

The cap has a fitting portion for the upper end opening of the inner container. Further, the fitting portion can be fitted into the upper end opening of the inner container to make the inside of the inner container liquid-tight. The number of caps may be one or plural. The cap may, for example, fit inside or outside the upper end opening of the inner container. For example, when the cap is attached to the upper end opening of the inner container, the cap can more reliably achieve a liquid-tight state of the inner container. is preferred. In this case, at least one of the inner fitting and the outer fitting is preferably threaded. As a specific example, the cap has, for example, a circular substrate, a first annular wall and a second annular wall. In this case, the first annular wall and the second annular wall are arranged concentrically on one surface of the circular substrate, the first annular wall being arranged inside and the second annular wall being arranged outside. are placed in The cap is fitted, for example, with the outer peripheral surface of the first annular wall and the inner peripheral surface of the inner container. Further, the cap is fitted with, for example, the inner peripheral surface of the second annular wall and the outer peripheral surface of the inner container.

It is preferable that the fitting portion of the cap is fitted into the upper end opening of the outer container so that the inside of the outer container can be kept liquid-tight. In this case, it can also be said that at least one of the inner diameter and outer diameter of the upper end opening of the outer container is the same as at least one of the inner diameter and outer diameter of the upper end opening of the inner container.

In the separation container kit of the present invention, the liquid-tight state of the collecting portion is achieved by the first press-contact portion, but is not limited to this. , a connecting (fixing) member that connects (fixes) the contact portions of the outer container and the inner container in a closely contacting state. As a specific example, the connecting member is, for example, an elastic member that covers contact portions of the locking portions in a state where the locking portions of the outer container and the inner container are locked and presses them in the contact surface direction. etc.

Hereinafter, as examples of the present invention, Embodiment 1 in which the first pressure contact portion is the tapered portion of the outer container and the inner container, Embodiment 2 in which the first pressure contact portion is a separate member, and the connecting member Embodiment 3 will be described with reference to FIGS. 1 to 7. FIG. In each figure, the same reference numerals are assigned to the same parts. In addition, in the drawings, for convenience of explanation, the structure of each part may be simplified as appropriate, and the dimensional ratio of each part may be schematically shown unlike the actual one. In addition, unless otherwise indicated, each description can be used for each embodiment.

(1) Embodiment 1
Embodiment 1 is an example of tapered portions of the outer container and the inner container, and will be described with reference to FIGS. 1 to 3. FIG. FIG. 1 is a schematic diagram showing an example of a separation container, (A) is a cross-sectional view of the outer container, (B) is a cross-sectional view of the inner container, and (C) is an inner container inside the outer container. FIG. 4 is a cross-sectional view of the inserted separation container; 2, (A) is a side view of the cap, (B) is a cross-sectional view of the cap in (A) taken along line AA, and (C) is a perspective view from below. FIG. 3 is a cross-sectional view of the separation container with the cap fitted to the upper end opening.

As shown in FIG. 1A, the outer container 10 has a bottomed cylindrical shape with an opening 106 at the upper end. , a connecting portion 103, a first inner peripheral tapered portion 104, and a tip portion 105, and a first threaded portion 108 is formed on the outer peripheral surface of the second inner peripheral tapered portion 101. As shown in FIG. As shown in FIG. 1B, the inner container 11 has a cylindrical shape with an upper end opening 117 and a lower end opening 118. From the top downward, the opening connecting portion 111, the second outer periphery It is composed of a tapered portion 112, a body portion 113, a connecting portion 114, a first outer peripheral taper portion 115, and a tip portion 116, and has a third threaded portion 119 on the outer peripheral surface of the opening connecting portion 111, and a second threaded portion 119. A cover portion 120 protrudes in an inverted L-shape from the outer peripheral surface of the outer peripheral taper portion 112 of , and a second threaded portion is formed on the inner peripheral surface of the cover portion 120 . The taper angle of the inner peripheral surface of the first inner peripheral tapered portion 104 and the taper angle of the outer peripheral surface of the first outer peripheral tapered portion 115 are substantially the same. The inner diameter of the lower end of the first inner peripheral tapered portion 104 is set smaller than the outer diameter of the lower end of the first outer peripheral tapered portion 115 . Also, the taper angle of the inner peripheral surface of the second inner peripheral tapered portion 101 and the taper angle of the outer peripheral surface of the second outer peripheral tapered portion 112 are substantially the same. The inner diameter of the lower end of the second inner peripheral tapered portion 101 is set smaller than the outer diameter of the lower end of the second outer peripheral tapered portion 112 . Then, as shown in FIG. 1C, by screwing together the first threaded portion 108 and the second threaded portion on the inner peripheral surface of the cover portion 120, the inner container is inserted into the outer container 10. 11 is inserted. In FIG. 1(C), when the inner container 11 is inserted into the outer container 10, the tip portion 105 of the outer container, the region below the lower end of the tip portion 116 of the inner container 11 and the inner portion of the tip portion 105 A region between the peripheral surface and the outer peripheral surface of the tip portion 116 serves as the collection portion 109 . The recovery unit 109 can recover, for example, sediments (separated substances) from inside the inner container 11 as will be described later. Further, when the inner container 11 is inserted into the outer container 10 , the outer peripheral surface of the first outer peripheral tapered portion 115 of the inner container 11 is aligned with the inner peripheral surface of the first inner peripheral tapered portion 104 of the outer container 10 . , and thereby the recovery portion 109 is brought into a liquid-tight state. Further, when the inner container 11 moves toward the bottom surface of the outer container 10 and the pressure contact between the inner peripheral surface of the first inner peripheral tapered portion 104 and the outer peripheral surface of the first outer peripheral tapered portion 115 is released, the inner container The outer peripheral surface of the second outer peripheral tapered portion 112 of 11 is in pressure contact with the inner peripheral surface of the second inner peripheral tapered portion 101 of the outer container 10 . As a result, the movement of gas between main body portion 102 and main body portion 113, between connection portion 103 and connection portion 114, and between first inner peripheral tapered portion 104 and first outer peripheral tapered portion 115 is prevented. Since it is regulated (airtight state), the recovery unit 109 is in a liquid-tight state.

As shown in FIGS. 2(A)-(C), the cap 20 has a circular substrate 21, a first annular wall 22 and a second annular wall 23. As shown in FIG. A first annular wall 22 is formed on the inner side and a second annular wall 23 is formed on the outer side concentrically on the lower surface of the circular substrate 21 . An annular groove is formed between the first annular wall 22 and the second annular wall 23 on the lower surface of the circular substrate 21 . The inner peripheral surface of the second annular wall 23 is formed with a fourth threaded portion to be threaded with the third threaded portion 119 of the inner container 11 . The first annular wall 22 is a fitting portion 22 that internally fits into the inner container 11 , and the second annular wall 23 is a fourth fitting portion 23 that externally fits (screws) into the inner container 11 . .

Attachment of the cap 20 to the separation container 1 comprising the inner container 11 and the outer container 10 will be described below. FIG. 3 shows the attachment of the cap to the separation container with the inner container 11 inserted inside the outer container 10 . A fourth threaded portion on the inner peripheral surface of the second annular wall 23 of the cap 20 is threaded with a third threaded portion 119 formed on the outer periphery of the opening 117 of the inner container 11 . At the same time, the outer peripheral surface of the first annular wall 22 of the cap 20 and the inner peripheral surface of the upper end portion of the inner container 11 are brought into contact to fit the first annular wall 22 of the cap 20 into the inner container 11 . Thereby, the cap 20 can be attached to the separation container 1, and the inside of the inner container 11 becomes liquid-tight.

In this embodiment, when the inner container 11 is inserted into the outer container 10, the inner peripheral surface of the first inner peripheral tapered portion 104 of the outer container 10 and the outer peripheral surface of the first outer peripheral tapered portion 115 of the inner container 11 is pressed against. As a result, the recovery unit 109 becomes liquid-tight. For this reason, when the separated liquid is introduced into the inner container 11, the recovery portion 109 will move between the inner peripheral surface of the outer container 10 and the outer peripheral surface of the inner container 11 (for example, between the main body portion 102 and the main body portion). 113, and between the connecting portion 103 and the connecting portion 114). Further, the outer container 10 and the inner container 11 are screwed together by the first screwing portion 108 and the second screwing portion of the cover portion 120 . Thus, by applying an axial force to the inner container 11, a larger pressure can be applied to the press-contact surfaces between the first inner peripheral tapered portion 104 and the first outer peripheral tapered portion 115. Therefore, the liquid-tight state of the recovery unit 109 can be achieved more reliably. Furthermore, when removing the inner container 11, it is possible to prevent the inner container 11 from being rapidly removed from the outer container 10. Therefore, the pressure from the inner container 11 to the collecting unit 109 caused by the negative pressure of the collecting unit 109 can be prevented. It can suppress the movement of the supernatant. As described above, according to the separation container kit of the present embodiment, it is possible to more reliably suppress an increase in the remaining amount of supernatant after removal of the inner container 11 .

The size of each member is not particularly limited. It suffices if pressure contact with the portion 115 is possible. The size of each member is exemplified below.

As for the size of the outer container 10, for example, the following conditions can be exemplified. The total volume is, for example, 0.5-100 mL, 1-50 mL. The internal height of the entire outer container 10 is, for example, 30 to 200 mm, 50 to 120 mm. The height of the second inner peripheral tapered portion 101 is, for example, 1 to 5 mm and 2 to 4 mm. The height of the body portion 102 is, for example, 80 to 120 mm and 105 to 115 mm. The height of the connecting portion 103 is, for example, 0 to 15 mm, 1 to 15 mm, 10 to 12 mm. The height of the first inner peripheral tapered portion 104 is, for example, 5 to 20 mm, 10 to 15 mm. The height of the tip portion 105 is, for example, 5 to 15 mm, 8 to 12 mm. The inner circumference diameter (inner diameter) of the opening 106 is, for example, 10 to 20 mm, 13 to 15 mm. The inner circumference diameter (inner diameter) of the upper end of the second inner circumference tapered portion 101 is, for example, 10 to 20 mm, 13 to 15 mm, 1 to 8 mm, and 5 to 6 mm. The inner circumference diameter (inner diameter) of the lower end of the second inner circumference tapered portion 101 is, for example, 10 to 20 mm, 14 to 16 mm. The inner circumference diameter (inner diameter) of the upper end of the first inner circumference tapered portion 104 is, for example, 8 to 18 mm, 10 to 13 mm. The inner circumference diameter (inner diameter) of the lower end of the first inner circumference tapered portion 104 is, for example, 2 to 10 mm, 5 to 7 mm. The diameter (inner diameter) of the inner periphery of the upper end of the collecting portion 109 is, for example, 5 to 15 mm, 6 to 7 mm. The capacity of the recovery unit 109 is, for example, 0.1-0.5 mL, 0.2-0.4 mL. The internal height of the recovery unit 109 is, for example, 4 to 10 mm and 5 to 8 mm.

As for the size of the inner container 11, for example, the following conditions can be exemplified. The total volume is, for example, 0.3-100 mL, 0.5-50 mL. The overall height of the inner container 11 is, for example, 60-100 mm, 80-90 mm. The height of the opening connecting portion 111 is, for example, 1 to 20 mm, 10 to 15 mm. The height of the second outer peripheral tapered portion 112 is, for example, 0 to 20 mm, 1 to 20 mm, or 10 to 15 mm. The height of the body portion 113 is, for example, 50-90 mm, 60-70 mm. The height of the connecting portion 114 is, for example, 0-20 mm, 1-20 mm, or 9-11 mm. The height of the first outer peripheral tapered portion 115 is, for example, 5 to 30 mm, 10 to 20 mm. The diameter of the inner periphery of the upper end opening 117 is, for example, 5 to 20 mm, 10 to 14 mm. The outer diameter (outer diameter) of the lower end of the connecting portion of the second inner peripheral tapered portion 112 to the cover portion 120 is, for example, 10 to 20 mm or 14 to 16 mm. The outer diameter (outer diameter) of the lower end of the second inner peripheral tapered portion 112 is, for example, 10 to 20 mm, 13 to 15 mm. The outer circumference diameter (outer diameter) of the upper end of the first outer circumference tapered portion 115 is, for example, 2 to 10 mm, 6 to 7 mm. The outer diameter (outer diameter) of the lower end of the first outer peripheral tapered portion 115 is, for example, 2 to 10 mm, 5 to 7 mm. The outer diameter (outer diameter) of the upper end of the tip portion 116 is, for example, 2 to 10 mm, 5 to 7 mm. The outer diameter (outer diameter) of the lower end of the tip portion 116 is, for example, 2 to 10 mm, 4 to 6 mm. The inner diameter (inner diameter) of the opening 118 at the lower end is, for example, 2 to 7 mm, 3 to 6 mm.

As for the size of the cap 20, for example, the following conditions can be exemplified. The outer diameter (outer diameter) of the circular substrate 21 is, for example, 10 to 25 mm, 15 to 22 mm. The thickness of the circular substrate 21 is, for example, 0.1 to 5 mm, 0.5 to 3 mm. The outer diameter (outer diameter) of the first annular wall 22 is, for example, 5 to 21 mm, 10 to 15 mm. The inner peripheral diameter (inner diameter) of the first annular wall 22 is, for example, 4 to 20 mm, 9 to 14 mm. The height of the first annular wall 22 is, for example, 1-10 mm, 2-8 mm. The outer diameter (outer diameter) of the second annular wall 23 is, for example, 11 to 25 mm, 15 to 22 mm. The inner peripheral diameter (inner diameter) of the second annular wall 23 is, for example, 10 to 22 mm, 13 to 16 mm. The height of the second annular wall 23 is, for example, 1-10 mm, 2-8 mm. The width of the annular groove between the first annular wall 22 and the second annular wall 23, that is, the distance between the outer peripheral surface of the first annular wall 22 and the inner peripheral surface of the second annular wall 23 is, for example, 0.5. ~5 mm, 0.8-3 mm.

A material for forming each member is not particularly limited. Examples of the material include synthetic resin materials, metal materials, and glass materials. Among these, for example, moldability, assemblability, processability of materials such as adhesiveness as necessary; sanitary aspects such as elution of additives into samples; functionality such as visibility; , said synthetic resin material is preferred. Examples of the material include fluororesin, polypropylene (PP), polyethylene (PE), polycarbonate (PE), polyester, polyurethane, polymethylpentene, methacryl, ABS (acrylonitrile-butadiene-styrene copolymer), polyethylene terephthalate (PET). ), polyvinyl chloride, silicone, ethylene vinyl acetate copolymer, synthetic rubber, synthetic resins such as various elastomers, among these, fluororesin, polypropylene (PP), polyethylene terephthalate (PET), polyethylene (PE) , polycarbonate (PC) are preferred. The fluororesin is preferably, for example, a hydrophobic fluororesin, particularly preferably polytetrafluoroethylene (PTFE).

Next, a method for separating sperm from a sperm-containing sample by centrifugation using the separation container kit of the present embodiment will be described with reference to FIGS. 1 to 3. FIG.

First, as shown in FIG. 1(C), the inner container 11 is arranged inside the outer container 10 . At this time, the inner peripheral surface of the first inner peripheral tapered portion 104 of the outer container 10 and the outer peripheral surface of the first outer peripheral tapered portion 115 of the inner container 11 are in pressure contact. As a result, the recovery unit 109 is in a liquid-tight state.

Next, a separation liquid is injected into the inner container 11 . A lower end of the inner container 11 has an opening 118 . Therefore, the separated liquid injected into the inner container 11 is introduced into the collecting portion 109 of the outer container 10 through the opening 118 . The amount of the separation liquid is not particularly limited, and can be appropriately determined according to the sizes of the outer container 10 and the inner container 11 . The separation liquid is, for example, 1-10 mL, 2-5 mL.

Next, a sample containing sperm is introduced into the inner container 11 . The sample is preferably layered on the separated liquid inside the inner container 11 .

Then, as shown in FIG. 3 , the fourth threaded portion on the inner peripheral surface of the second annular wall 23 of the cap 20 is connected to the third threaded portion 119 formed on the outer periphery of the opening 117 of the inner container 11 . screw together. At the same time, the outer peripheral surface of the first annular wall 22 of the cap 20 and the inner peripheral surface of the upper end portion of the inner container 11 are brought into contact to fit the first annular wall 22 of the cap 20 into the inner container 11 . This makes the inside of the inner container 11 liquid-tight. In addition, liquids such as a separation liquid are not shown in the figure (the same applies hereinafter). Then, the separation container 1 is subjected to centrifugation. Centrifugation is usually performed by setting the separation container in a centrifugal rotor. At this time, if there is a gap between the set separation container 1 of this embodiment and the centrifugal rotor, an adjuster may be attached to the separation container 1 . Conditions for centrifugation are not particularly limited, and are, for example, 1000 G (1000×9.80665 m/s ² ) for 20 to 30 minutes. Sperm are separated from the sample by centrifugation, and the sperm are precipitated (separated) from the opening 118 of the inner container 11 to the collecting portion 109 of the outer container 10 .

After centrifugation, the inner container 11 is removed from the outer container 10 with the cap 20 attached. The interior of the inner container 11 contains unnecessary matter of the sample. However, by attaching the cap 20, the inside of the inner container 11 is in a liquid-tight state. The outer container 10 and the inner container 11 are screwed together by the first screwing portion 108 and the second screwing portion of the cover portion 120 . Therefore, the inner container 11 can be gently removed from the outer container 10 without leakage of liquid from the inside of the inner container 11 . Then, the separation liquid containing the sedimented sperm is recovered in the recovery section 109 of the outer container 10 .

The separation liquid is not particularly limited, and can be determined as appropriate according to, for example, the types of the sample and separation target. Examples of the separation liquid include a density gradient carrier, a medium, and a buffer solution. The density gradient carrier can also be called, for example, a specific gravity adjuster. The density gradient carrier is not particularly limited, and examples thereof include percoll, modified colloidal silica, sucrose polymer, ficoll and the like. The Percoll is usually a colloidal silica sol with a polyvinylpyrrolidone coating. The Percoll is preferably, for example, Percoll that has been made isotonic by adding the medium after removing endotoxin. The concentration of Percoll is not particularly limited, and is preferably 90 to 98%, for example. For example, when the object to be separated is sperm, the medium may be a HEPES-containing liquid or the like.

The heights of the outer container 10 and the inner container 11 are not particularly limited, and can be appropriately set according to their respective capacities. For the outer container 10 and the inner container 11, for example, the height of the outer container 10 relative to the height of the inner container 11 is compared with the height of the outer container 10 and the inner container 11 shown in FIGS. , can be higher or lower. As a specific example, the height of the outer container 10 is compared with the height of the outer container 10 shown in FIGS. 1A to 1C, as shown in FIGS. The height with respect to the vessel 11 may be lowered. As a result, the separation container 1 is temporarily press-contacted between the inner peripheral surface of the first inner peripheral tapered portion 104 and the outer peripheral surface of the first outer peripheral taper portion 115, which are the first press-contact portion, and the second press-contact portion. Even if pressure contact between the second inner peripheral tapered portion 101 and the outer peripheral surface of the second outer peripheral tapered portion 112 is released, the volume of the space between the outer container 10 and the inner container 11 can be reduced. Therefore, it is possible to suppress an increase in the remaining amount of the supernatant after the removal of the inner container.

In the separation container of Embodiment 1, the lower end of the inner container 11 is an opening, but as shown in FIG. The one-way valve 124 is, for example, a valve that allows liquid to move only from the inside to the outside (downward) of the inner container 11 due to pressure changes such as centrifugal separation. When the inner container 11 is provided with the one-way valve 124, the outer container 10 is recovered from the inner container 11 via the one-way valve 124 only while the centrifugal separation is being performed with the inner container 11 inserted into the outer container 10. A liquid can be introduced into the portion 109 . Even if the inner container 11 is removed from the outer container 10 after centrifugation, the one-way valve 124 prevents the liquid from moving from the opening 118 of the inner container 11 to the collecting portion 119 . For this reason, it is possible to more reliably suppress an increase in the remaining amount of the supernatant after the removal of the inner container.

Note that the above-described additional mode can also be applied, for example, to Embodiments 2 and 3 to be described later.

(2) Embodiment 2
Embodiment 2 is an example using the separate member, and will be described with reference to FIG. FIG. 6 is an example of a separation container using the separate member, (A) is a partially enlarged cross-sectional view around the tip portion 105 of the outer container 10, and (B) and (C) are the outer container 10. It is a partially enlarged cross-sectional view around the upper end.

As shown in FIG. 6A, in the separation container 1c of Embodiment 2, the inner container 11 does not have the first outer peripheral tapered portion 115 and the tip portion 116, and the lower end of the connection portion 114 has a cylindrical tip portion. 121 are formed. Further, an O-ring 122a, which is a sealing member, is arranged on the outer peripheral surface of the lower end of the distal end portion 121. As shown in FIG. Except for this point, the separation container 1c of Embodiment 2 has the same configuration as the separation container 1 of Embodiment 1, and the description thereof can be used. The outer peripheral surface of the tip portion 121 may or may not contact the inner peripheral surface of the outer container 10, for example. In the separation container 1c, when the inner container 11 is inserted into the outer container 10, the O-ring 122a is positioned between the outer peripheral surface of the lower end of the tip portion 121 and the inner peripheral surface of the outer container 10 (for example, the first inner peripheral taper). inner peripheral surface of the portion 104). Therefore, the inner peripheral surface of the outer container 10 and the outer peripheral surface of the inner container 11 are indirectly pressure-contacted by the O-ring 122a. Thus, the recovery unit 109 becomes liquid-tight. Therefore, according to the separation container kit of the present embodiment, it is possible to suppress an increase in the remaining amount of supernatant after removal of the inner container 11 .

In the separation container 1c of Embodiment 2, the O-ring 122a is arranged on the outer peripheral surface of the lower end of the tip portion 121, but the arrangement of the sealing member such as the O-ring 122a is not limited to this. The sealing member is, for example, a contact portion between the end surface of the opening 106 of the outer container 10 and the inner container 11, that is, the outer peripheral surface of the inner container 11 and the cover portion, as indicated by reference numeral 122b in FIG. 120 may be located in the groove. Also, the sealing member may be arranged on the outer peripheral surface of the inner container 11 corresponding to the open end of the outer container 10, as indicated by reference numeral 122c in FIG. 6(C), for example.

(3) Embodiment 3
Embodiment 3 is an example using a connecting member, and will be described with reference to FIG. FIG. 7 is an example of a separation container using the connecting member, and is a partially enlarged cross-sectional view around the upper end of the outer container 10. As shown in FIG.

As shown in FIG. 7, in the separation container 1f of Embodiment 3, the outer container 10 has a first engaging portion 110 projecting radially from the outer peripheral surface in place of the first threaded portion 108. , the inner container 11 has a second locking portion 123 that protrudes radially from the outer peripheral surface and is locked by the first locking portion 110 instead of the cover portion. In addition, the contact surfaces of the first locking portion 110 and the second locking portion 123 are controlled by the connecting member 30 that covers the projecting portions of the first locking portion 110 and the second locking portion 123 . It is fixed in a pressed state. Except for this point, the separation container 1f of Embodiment 3 has the same configuration as the separation container 1 of Embodiment 1, and the description thereof can be used. The connecting member 30 may cover the whole of the first locking portion 110 and the second locking portion 123, or may cover a part of them. In the separation container 1f, the contact surfaces of the first locking portion 110 and the second locking portion 123 are fixed by the connecting member 30 in a state of being pressed against each other. As a result, between the second inner peripheral tapered portion 101 and the second outer peripheral tapered portion 112, between the main body portion 102 and the main body portion 113, between the connecting portion 103 and the connecting portion 114, and between the first inner peripheral taper portion Since movement of gas between the peripheral tapered portion 104 and the first peripheral tapered portion 115 is restricted (airtight state), the collection portion 109 is liquid-tight. Therefore, according to the separation container kit of the present embodiment, it is possible to suppress an increase in the remaining amount of supernatant after removal of the inner container 11 .

(4) Embodiment 4
Embodiment 4 is an example provided with a temporary fixing portion, and will be described with reference to FIG. 8 . FIG. 8 is an example of the separation container provided with the temporary fixing portion, (A) is a partially enlarged side view of the periphery of the opening 106 of the outer container 10g, and (B) is the cover portion 120 of the inner container 11g. 1 is a partially enlarged cross-sectional view of FIG.

As shown in FIG. 8(A), a separation container 1g of Embodiment 4 has a first groove formed by ribs 125a and 125b, which are two convex portions, in a groove of a first screw-engagement portion 108 of an outer container 10g. has a temporary fixing portion 125 of . As shown in FIG. 8A, ribs 125a and 125b are spaced apart in the groove. Further, as shown in FIG. 8(B), in the separation container 1g of the fourth embodiment, the cover portion 120 of the inner container 11g is formed as a threaded portion that can move through the groove of the first threaded portion 108. 2 temporary fixing portions 126 are provided. The circumferential length of the second temporary fixing portion 126 (screw portion) is set narrower than the circumferential interval between the ribs 125a and 125b. Except for these points, the separation container 1g of Embodiment 4 has the same configuration as the separation container 1 of Embodiment 1, and the description thereof can be used.

In the separation container 1g, the first threaded portion 108 of the outer container 10g and the second threaded portion of the cover portion 120 of the inner container 11g are screwed together, and the second temporary fixing portion 126 is connected to the first threaded portion. When moved along the groove of the threaded portion 108, the second temporary fixing portion 126 comes into contact with the rib 125b. In this state, when the inner container 11g is further screwed into the outer container 10g, the second temporary fixing portion 126 climbs over the rib 125b and is arranged between the rib 125a and the rib 125b. Then, in order for the second temporary fixing portion 126 to separate from the ribs 125a and 125b that are the first temporary fixing portion 125, a force is required to overcome the ribs 125a and 125b. For this reason, the second temporary fixing portion 126 is not attached to the first temporary fixing portion 125 unless a force for detaching the inner container 11g from the outer container 10g or a force for fastening the inner container 11g to the outer container 10g is applied. It will be in a held state, that is, a temporarily fixed state. When a force is applied to fasten the inner container 11g to the outer container 10g in the temporarily fixed state, the second temporary fixing portion 126 climbs over the rib 125a, and the inner container 11g moves toward the bottom surface of the outer container 10g. It moves and the 1st threaded part 108 and the 2nd threaded part fasten. As a result, the first outer tapered portion 115 and the first inner tapered portion 104 are brought into pressure contact.

In the separation container 1g, when the inner container 11g and the outer container 10g are temporarily fixed, the inner container 11g is not fastened to the outer container 10g. The inner container 11g is temporarily fixed in a relatively upward direction (opening 106 side) as compared with the case where the inner peripheral tapered portion 104 of 1 is in pressure contact. Therefore, in a state where the first temporary fixing portion 125 and the second temporary fixing portion 126 are engaged and the inner container 11g is temporarily fixed inside the outer container 10g, the first outer peripheral tapered portion 115, Pressure contact with the first inner peripheral tapered portion 104 does not occur, that is, the entire contact surfaces of both are not in contact. Therefore, according to the separation container 1g, when the separation container 1g is stored in a temporarily fixed state, the first outer peripheral tapered portion 115 and the first inner peripheral tapered portion 104 are pressed against each other. , it is possible to prevent the inner container 11g from conforming to the outer container 10g, that is, from deforming the outer peripheral shape of the inner container 11g to match the inner peripheral shape of the outer container 10g. Therefore, according to the separation container 1g, since the pressure contact between the first outer peripheral tapered portion 115 and the first inner peripheral tapered portion 104 can be ensured during use, the liquid-tight state of the recovery portion 109 can be further improved. can be assured. Further, according to the separation container 1g, by storing the separation container 1g in a temporarily fixed state, the first outer peripheral tapered portion 115 and the first inner peripheral tapered portion 104 are pressed against each other. In comparison, since the stress applied to the inner container 11g and the outer container 10g during storage can be suppressed, the possibility of damage such as cracks occurring during storage or use can be reduced.

In the separation container of Embodiment 4, one set of the first temporary fixing portion 125 and the second temporary fixing portion 126 is formed, but a plurality of sets may be formed.

In the separation container 1g of Embodiment 4, the temporary fixing portions are formed in the first threaded portion 108 and the second threaded portion, but may be arranged in other portions of the separation container 1g. As a specific example, the temporary fixing portion may be formed, for example, around the opening 106 of the outer container 11g.

In the separation container 1g of Embodiment 4, ribs 125a and 125b are formed as the first temporary fixing portion 125, and screw portions are formed as the second temporary fixing portion 126. However, the first temporary fixing portion The configuration of 125 and the configuration of the second temporary fixing portion 126 may be reversed. That is, a threaded portion may be formed as the first temporary fixing portion 125 and two ribs may be formed as the second temporary fixing portion 126 . This point is the same for other configurations of the temporary fixing portions to be described later.

In the separation container 1g of Embodiment 4, the form of the temporary fixing portion may be another form. As a specific example, the first temporary fixing portion 125 is unevenness formed in the groove of the first screwing portion 108, and the second temporary fixing portion 126 is a corresponding groove formed in the second screwing portion. It is uneven. In this case, the concave portion of the first temporary fixing portion 125 is fitted with the convex portion of the second temporary fixing portion 126, and the convex portion of the first temporary fixing portion 125 is fitted with the concave portion of the second temporary fixing portion 126. Mates with One or a plurality of the uneven structures may be provided. Further, the unevenness may be, for example, a wave shape.

The first temporary fixing portion 125 is a concave portion formed in the groove of the first threaded portion 108, and the second temporary fixing portion 126 is a corresponding convex portion formed in the second threaded portion. Department. In this case, the concave portion of the first temporary fixing portion 125 fits with the convex portion of the second temporary fixing portion 126 . The number of the concave portion and the convex portion may be one, or a plurality thereof may be provided.

In each embodiment, separation of sperm from a sample is exemplified as an application of the separation container kit of the present invention, but this is an example and the present invention is not limited to this application. When the separation container kit of the present invention is used to separate sperm from a sample, the separation method is not particularly limited, and can be applied to, for example, density gradient centrifugation, swim-up method, and the like. In addition, it can be used, for example, for separation of cell organelles, separation of DNA or RNA, separation of specific components of blood (eg, red blood cells, mononuclear cells, plasma), and the like. The sample is not particularly limited, and may be a sample containing the object to be separated, and specific examples thereof include semen, blood, and the like.

Next, examples of the present invention will be described. However, the present invention is not limited by the following examples.

(1) Production of Separation Container Kit As the separation container kit of the example, the separation container kit of Embodiment 1 was produced by injection molding. In the separation container kit of the example, the first inner peripheral tapered portion 104 had an inner diameter of 6.4 mm at the lower end and was formed with a 6% taper. The first outer peripheral tapered portion 115 has an inner diameter of 6.4 mm at the lower end and is tapered by 6%. The outer diameter of the lower end of the tip portion 116 was set to 6 mm, and the outer peripheral surface was formed so as not to come into contact with the outer container 10 . The volume of the collection part 109 of the separation kit of the example is 0.25±0.05 mL.

In the separation container kit of the comparative example, the outer container 10 did not form the first inner peripheral tapered portion 104 and the inner container 11 did not form the first outer peripheral tapered portion 115 and the tip portion 116 . In the separation container kit of the comparative example, the first engaging portion is formed instead of the first threaded portion 108 of the outer container 10, and the first engaging portion is formed instead of the cover portion 120 of the inner container 11. A second engaging portion corresponding to the stop portion was formed, and the second engaging portion of the inner container 11 was engaged with the first engaging portion of the outer container 10 . Except for this point, it was the same as the separation container kit 1 of the above example. The volume of the collection part 109 of the separation container kit of the comparative example is the same as that of the separation kit 1 of the example.

(2) Measurement of Remaining Volume of Supernatant It was placed by inserting the inner container 11 into the outer container 10 of the separation container kit of Examples (n=3, Examples 1 to 3). As a result, the first inner peripheral tapered portion 104 and the first outer peripheral tapered portion 115 are brought into pressure contact, and the recovery portion 109 is liquid-tight. Next, 8 mL of water was introduced into the inner container 11 and the cap 20 was attached to the upper opening of the inner container 11 . After the mounting, the separation container kit 1 was centrifuged twice under conditions of 700×g for 30 minutes. Then, the inner container 11 was removed, and the remaining amount of the supernatant in the outer container 10 was measured. In addition, using the separation container kit of the comparative example (n=3, Comparative Examples 1 to 3), the remaining amount of the supernatant was similarly measured. These results are shown in Table 1 below.

As shown in Table 1 above, the separation container kit of the example was able to reduce the remaining amount of the supernatant to about ⅓ of that of the separation container kit of the comparative example. From these results, it was found that the separation container kit of the present invention can suppress an increase in the remaining amount of the supernatant after removal of the inner container. Therefore, according to the separation container kit of the present invention, it was found that the concentration of the substance to be separated after separation can be increased.

Although the present invention has been described with reference to the embodiments and examples, the present invention is not limited to the above embodiments and examples. Various changes that can be understood by those skilled in the art can be made to the configuration and details of the present invention within the scope of the present invention.

According to the separation container kit of the present invention, it is possible to suppress an increase in the remaining amount of supernatant after removal of the inner container. Therefore, according to the separation container kit of the present invention, for example, the concentration of the separation target after separation can be increased. Therefore, the separation container kit of the present invention can be said to be a very useful tool in the separation of objects to be separated such as sperm.

1, 1a, 1b, 1c, 1d, 1e, 1f separation container 10, 10a, 10b, 10c, 10d, 10e, 10f, 10g outer container 11, 11a, 11b, 11c, 11d, 11e, 11f, 11g inner container 101 Second inner peripheral tapered portions 102, 113 Body portions 103, 114 Connecting portion 104 First inner peripheral tapered portions 105, 116, 121 Tip portions 106, 117, 118 Opening portion 108 First threaded portion 109 Recovery portion 110 First locking portion 111 Opening connection portion 112 Second outer tapered portion 115 First outer tapered portion 119 Third threaded portion 120 Cover portions 122a, 122b, 122c O-ring 123 Second locking portion 124 One side Valve 125 First temporary fixing portion 125a, b Rib 126 Second temporary fixing portion (screw portion)
20 cap 21 circular substrate 22 first annular wall 23 second annular wall 30 connecting portion

Claims (13)

including a separate container and cap;
The separation container is
having an outer container, an inner container, and a first pressure contact part,
The outer container is
It has a bottomed cylindrical shape with an open upper end and a closed lower end,
The inner container is
It has a cylindrical shape with an upper end and a lower end opened,
The inner container and the outer container are
The inner container can be inserted into the outer container in a state in which the respective vertical directions are aligned,
A space between the lower end of the outer container and the lower end of the inner container in a state where the inner container is inserted into the outer container is a separation target object recovery section,
The first pressure contact portion is
arranged or formed between the inner peripheral surface of the outer container and the outer peripheral surface of the inner container,
The cap is
Having a fitting portion for the upper end opening of the inner container,
The fitting portion can make the inside of the inner container liquid-tight by fitting into the upper end opening of the inner container,
In a state in which the inner container is inserted into the outer container, the inner peripheral surface of the outer container and the outer peripheral surface of the inner container are pressed together by the first pressure contact portion,
A separation container kit for separating an object to be separated from a liquid sample, characterized in that the recovery section is brought into a liquid-tight state by the pressure contact. The separation container is
a moving means capable of moving the inner container toward the bottom surface of the outer container;
2. The separation container kit according to claim 1, wherein said moving means inserts said inner container into said outer container. The outer container is
Having a first threaded portion for the inner container,
The inner container is
having a second threaded portion for the first threaded portion;
When the first screwing portion and the second screwing portion are screwed together and the inner container is inserted into the outer container, the inner peripheral surface of the outer container and the outer peripheral surface of the inner container 3. The separation container kit according to claim 1 or 2, wherein and are press-contacted by said first press-contact portion. The outer container is
Having a first temporary fixing portion for temporarily fixing the inner container,
The inner container is
Having a second temporary fixing portion for the first temporary fixing portion,
In a state where the first temporary fixing portion and the second temporary fixing portion are engaged and the inner container is temporarily fixed inside the outer container, the inner peripheral surface of the outer container and the inner container 4. The separation container kit according to any one of claims 1 to 3, wherein the outer peripheral surface is not press-contacted by the first press-contact portion. The outer container is
Having a first temporary fixing portion for temporarily fixing the inner container,
The first temporary fixing portion is formed in the first threaded portion,
The inner container is
Having a second temporary fixing portion for the first temporary fixing portion,
The second temporary fixing portion is formed in the second threaded portion,
When the first screwing portion and the second screwing portion are screwed together and the inner container is inserted into the outer container, the first temporary fixing portion and the second temporary fixing portion are engaged. are engaged and the inner container is temporarily fixed inside the outer container, the inner peripheral surface of the outer container and the outer peripheral surface of the inner container are pressed against each other by the first pressure contact portion. figure,
When the first threaded portion and the second threaded portion are further screwed together, the engagement between the first temporary fixing portion and the second temporary fixing portion is released, and the first and said second threaded portion are engaged, and the inner peripheral surface of said outer container and the outer peripheral surface of said inner container are press-contacted by said first press-contact portion. Separation container kit. One of the first temporary fixing portion and the second temporary fixing portion is two convex portions spaced apart from each other in the groove of the first threaded portion or the second threaded portion,
6. The separation container kit according to claim 5, wherein the other is a protrusion that can be locked between the two protrusions. the first pressure contact portion is a pair of corresponding tapered portions of the outer container and the inner container;
The outer container has a first inner peripheral tapered portion on the inner peripheral surface,
The inner container has a first outer peripheral tapered portion on the outer peripheral surface,
Direct contact between the inner peripheral surface of the first inner peripheral tapered portion and the outer peripheral surface of the first outer peripheral tapered portion causes the inner peripheral surface of the outer container and the outer peripheral surface of the inner container to , are crimped together. At a corresponding portion between the first inner peripheral tapered portion and the first outer peripheral tapered portion,
The ratio of the inner diameter (I) of the first inner peripheral tapered portion to the outer diameter (O) of the first outer peripheral tapered portion is in the range of 1:1.0003 to 1:1.03. Item 8. The separation container kit according to item 7. Separation container kit according to claim 7 or 8, wherein the height (LT1) of said first inner circumferential taper is in the range of 1 to 5 mm. The first pressure contact portion is
10. The separation container kit according to any one of claims 1 to 9, arranged so as to be adjacent to the collection part in a state in which the inner container is inserted into the outer container. The inner container is
It has a cylindrical tip at the lower end,
The tip is
The separation container kit according to any one of claims 1 to 10, wherein the outer peripheral surface of the inner container does not contact the inner peripheral surface of the outer container when the inner container is inserted into the outer container. having a second pressure contact portion;
The second pressure contact portion is arranged or formed between the inner peripheral surface of the outer container and the outer peripheral surface of the inner container,
When the inner peripheral surface of the outer container and the outer peripheral surface of the inner container are in pressure contact with the first pressure contact portion in a state in which the inner container is inserted into the outer container, The inner peripheral surface and the outer peripheral surface of the inner container are not press-contacted by the second press-contact portion,
When the inner container moves toward the bottom surface of the outer container, and furthermore, the pressure contact between the inner peripheral surface of the outer container and the outer peripheral surface of the inner container by the first pressure contact portion is released, the outer container The inner peripheral surface and the outer peripheral surface of the inner container are press-contacted by the second press-contact portion,
12. The separation container kit according to any one of claims 1 to 11, wherein the recovery part is brought into a liquid-tight state by the pressure contact of the second pressure contact part. 13. The separation container kit according to any one of claims 1 to 12, wherein the fitting part can make the inside of the outer container liquid-tight by fitting into the upper end opening of the outer container.

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