Filling cap and method

Abstract

The purpose of the present invention is to realize an efficient liquid-dispensing cap having a simple construction. This liquid-dispensing cap is to be used by being mounted to a liquid dispensing container, and comprises: a lid body that is detachably mounted to a liquid dispensing container; and a liquid-dispensing tube and an air intake tube that respectively extend upward from the lid body. A stepped portion is provided in a flow path of the air intake tube.

Description

[Technical Field]

[0001] The present invention relates to a liquid filling cap and method. [Background technology]

[0002] In recent years, attempts have been made to transplant various cells to repair damaged tissues, etc. For example, attempts have been made to use fetal cardiomyocytes, skeletal myoblasts, mesenchymal stem cells, cardiac stem cells, ES cells, cardiomyocytes, etc. to repair myocardial tissue damaged by ischemic heart diseases such as angina pectoris and myocardial infarction. As part of these attempts, cell structures formed using scaffolds and sheet-like cell cultures in which cells are formed into a sheet have been developed.

[0003] These cell cultures have traditionally been produced manually by workers with specialized knowledge in clean rooms called cell processing centers (CPCs), and the production costs and labor required for such cell cultures are high, so there is a need for greater efficiency. Therefore, an automated cell culture device has been proposed that uses an articulated robot to perform the tasks related to the culture of these cells (Patent Document 1).

[0004] In order to speed up the liquid injection work in the above-mentioned automatic culture, the present inventors have proposed a device that is attached to a liquid injection container (Patent Document 2). This device includes a lid that can be detachably attached to the liquid injection container, a liquid injection tube that can be fitted into a first through-hole provided in the lid, and an air intake tube that can be fitted into a second through-hole provided in the lid, and is configured to allow air from outside the liquid injection container to pass through but not liquid from inside the liquid injection container by providing a check valve in the air intake tube. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] International Publication No. 2016 / 104666 [Patent Document 2] International Publication No. 2019 / 230922 Summary of the Invention [Problem to be solved by the invention]

[0006] In developing a means for speeding up the liquid injection process, the inventors encountered problems such as unstable injection volume when the liquid injection process was repeated. Therefore, an object of the present invention is to solve such problems and to realize a liquid injection cap with a simple configuration and a rational design. [Means for solving the problem]

[0007] In the course of intensive research to solve the above problems, the inventors of the present invention have noticed that the unstable injection amount is caused by liquid getting into the intake tube (clogging), unstable behavior of the check valve, etc. As a result of further research, they have found that the above problems can be solved by providing a step in the flow path of the intake tube, and have completed the present invention.

[0008] That is, the present invention relates to the following. [1] A liquid injection cap to be attached to a liquid injection container, said liquid injection cap comprising a lid body that can be attached and detached to the liquid injection container, a liquid injection tube and an air intake tube extending upward from the lid body, and a step portion provided in the flow path of the air intake tube. [2] The liquid filling cap according to [1], wherein the inner diameter of the intake hole of the intake tube is smaller than the inner diameter of the intake tube. [3] The liquid filling cap according to [1] or [2], wherein the inner diameter of the intake hole of the intake tube is preferably 0.5 mm to 3.0 mm, more preferably 1.0 mm to 2.0 mm, even more preferably 1.1 mm to 1.6 mm, and most preferably 1.2 mm to 1.4 mm. [4] The liquid filling cap according to any one of [1] to [3], wherein the inner diameter of the intake tube is preferably 2.0 mm to 10.0 mm, more preferably 2.0 mm to 7.0 mm, and even more preferably 2.0 mm to 4.0 mm. [5] The liquid filling cap according to any one of [1] to [4], wherein the length of the intake tube extending from the lid body is preferably 5 to 100 mm, more preferably 10 to 50 mm, and even more preferably 10 to 20 mm.

[0009] [6] The liquid filling cap according to any one of [1] to [5], further comprising an extension tube that extends the intake tube downward from the lid body. [7] The liquid injection cap according to any one of [1] to [6], wherein the inner diameter of the extension tube is preferably 2.0 mm to 10.0 mm, more preferably 2.0 mm to 7.0 mm, and even more preferably 2.0 mm to 4.0 mm. [8] The liquid filling cap according to any one of [1] to [7], wherein the length of the extension tube is preferably 5 to 100 mm, more preferably 10 to 50 mm, and even more preferably 15 to 30 mm. [9] A method for injecting a liquid, comprising: (a) preparing an injection container containing a liquid; (b) attaching an injection cap to the injection container, the injection cap including a lid that can be removably attached to the injection container, an injection tube that extends upward from the lid, and an intake tube, with a step provided in the flow path of the intake tube; (c) tilting the injection container downward to inject the liquid through the injection tube; and (d) releasing the downward tilt of the injection container to terminate the injection.

[10] The method according to [9], wherein the inclination of the infusion container is carried out by rotating the infusion container around a predetermined axis.

[11] The method according to [9] or

[10] , further comprising step (e) of repeating steps (c) and (d) to perform multiple injection operations consecutively.

[12] (f) The method according to any one of [9] to

[11] , further comprising the step of determining the amount of liquid injected from the injection time.

[13] The method according to any one of [9] to

[12] , wherein the liquid injection cap further includes an extension tube that extends the intake tube downward from the lid body. [Effects of the Invention]

[0010] According to the present invention, it is possible to prevent the amount of liquid injected from becoming unstable, and to perform efficient, rapid, and highly accurate continuous liquid injection work. Furthermore, since the device has a simple structure that does not use a check valve, it can be manufactured inexpensively. According to the present invention, the liquid injection operation can be performed by accurately aiming at the storage container, and the occurrence of liquid dripping can be minimized, making it suitable for producing cell cultures in clean rooms, etc. Furthermore, by increasing the liquid injection speed, clogging of the intake tube can be more reliably prevented. [Brief explanation of the drawings]

[0011] [Figure 1] FIG. 1 is a schematic diagram of a liquid injection cap 1 according to a first embodiment of the present invention. [Figure 2] FIG. 2 is a schematic diagram illustrating the liquid injection operation using the liquid injection cap 1 of FIG. [Figure 3] FIG. 3 shows a schematic diagram of a modified example of the liquid injection cap 1. In FIG. [Figure 4] FIG. 4 is a schematic diagram of a liquid injection cap 1' according to a second embodiment of the present invention. [Figure 5] FIG. 5 is a schematic diagram illustrating the liquid injection operation using the liquid injection cap 1' of FIG. [Figure 6] FIG. 6 is a schematic diagram illustrating the principle of the liquid injection shown in FIG. DETAILED DESCRIPTION OF THE INVENTION

[0012] Examples of components constituting the liquid in the present invention include water, physiological saline, physiological buffer solutions (e.g., HBSS, PBS, EBSS, Hepes, sodium bicarbonate, etc.), culture media (e.g., DMEM (DME), MEM, F12, DMEM / F12, RPMI1640, MCDB, L15, SkBM, RITC80-7, IMDM, etc.), sugar solutions (sucrose solution, Ficoll-paque (registered trademark) PLUS, etc.), seawater, serum-containing solutions, Renografin (registered trademark) solution, metrizamide solution, meglumine solution, glycerin, ethylene glycol, ammonia, benzene, toluene, acetone, ethyl alcohol, benzol, oil, mineral oil, animal fat, vegetable oil, olive oil, colloidal solution, liquid paraffin, turpentine oil, linseed oil, and castor oil.

[0013] The storage container in the present invention is not particularly limited, but examples thereof include cell culture containers, cell culture flasks for adherent cells, cell culture flasks for suspension cells, etc. A cell culture flask is a container that has a roughly rectangular main body, at least one flat surface of the main body is subjected to a surface treatment required for cell culture, and multiple flasks can be stacked with the cell culture surface facing downwards to enable multi-stage culture. The liquid-injecting container in the present invention is not particularly limited as long as it is a container that can contain the culture medium to be injected into the storage container, but examples include shaker flasks, Erlenmeyer flasks, roller bottles, liquid-injecting bottles, beakers, culture medium bottles, square culture medium bottles, sterilized bottles, and the like.

[0014] The robot in the present invention is not particularly limited, but examples thereof include linear motion / rotation devices, manipulators, articulated robots, etc. Examples of articulated robots include two-axis articulated robots, three-axis articulated robots, four-axis articulated robots, five-axis articulated robots, six-axis articulated robots, and seven-axis articulated robots.

[0015] In the present invention, the "predetermined axis" refers to the axis that serves as the center of rotation when rotating the infusion container, and in the case where the infusion container is a typical vertically long container, the predetermined axis is set as an axis perpendicular to the long axis of the container. In this invention, "TCP" refers to the Tool Center Point, which is a coordinate system for expressing the position and posture of a controlled object such as a tool, gripper, or workpiece located at the tip of a robot. The TCP can be set to any position or posture (convenient for operation and control) of, for example, an end effector (gripper, tool, etc.) or workpiece (flask, bottle, etc.), and in the case of a six-axis articulated robot, it is usually defined with respect to the coordinate system of the sixth axis of the robot.

[0016] In the present invention, "rotating around a predetermined axis" refers to rotating an object around a predetermined axis. For example, if the predetermined axis is set at one end of the opening of the liquid injection container, the liquid inside the liquid injection container can be discharged simply by rotating around that end. Furthermore, even if the predetermined axis is set at the center (center of gravity) of the liquid injection container, the liquid injection container can be rotated around one end of the opening of the liquid injection container as described above by combining a rotational movement around the central axis of the liquid injection container with a translational movement along an arc orbit.

[0017] Furthermore, for example, if the robot is an articulated robot, the efficiency of robot control can be improved by associating a predetermined axis with the TCP. If the robot is, for example, a six-axis articulated robot, the rotation axis of the sixth axis and the rotation axis of the TCP can be made parallel, so that the liquid injection container can be rotated as described above by rotating the sixth axis and slightly moving the first to fifth axes. Furthermore, if the rotation axis of the sixth axis and the rotation axis of the TCP are made to coincide, the liquid injection container can be rotated as described above by rotating the sixth axis alone.

[0018] Examples of cells in the present invention include, but are not limited to, adherent cells (adherent cells). Adherent cells include, for example, adherent somatic cells (e.g., cardiac myocytes, myoblasts, fibroblasts, epithelial cells, endothelial cells, hepatocytes, pancreatic cells, kidney cells, adrenal cells, periodontal ligament cells, gingival cells, periosteal cells, skin cells, synovial cells, chondrocytes, etc.) and stem cells (e.g., tissue stem cells such as mesenchymal stem cells and cardiac stem cells, embryonic stem cells, pluripotent stem cells such as iPS (induced pluripotent stem) cells, etc.). Somatic cells may be differentiated from stem cells, particularly iPS cells. Non-limiting examples of cells capable of forming sheet-shaped cell cultures include myoblasts (e.g., skeletal myoblasts), mesenchymal stem cells (e.g., derived from bone marrow, adipose tissue, peripheral blood, skin, hair roots, muscle tissue, endometrium, placenta, and umbilical cord blood), cardiomyocytes, fibroblasts, cardiac stem cells, embryonic stem cells, iPS cells, synovial cells, chondrocytes, epithelial cells (e.g., oral mucosal epithelial cells, retinal pigment epithelial cells, nasal mucosal epithelial cells), endothelial cells (e.g., vascular endothelial cells), hepatocytes (e.g., hepatic parenchymal cells), pancreatic cells (e.g., pancreatic islet cells), kidney cells, adrenal cells, periodontal ligament cells, gingival cells, periosteal cells, and skin cells. In the present invention, cells that form monolayer cell cultures, such as myoblasts or cardiomyocytes, are preferred, with skeletal myoblasts or cardiomyocytes derived from iPS cells being particularly preferred.

[0019] Preferred embodiments of the present invention will now be described in detail with reference to the drawings. [First embodiment] One aspect of the present invention relates to a liquid injection cap that is attached to a liquid injection container, and includes a lid body that can be attached and detached to the liquid injection container, and a liquid injection tube and an air intake tube that extend upward from the lid body, and the liquid injection cap has a step portion in the flow path of the air intake tube.

[0020] First, a first embodiment of the present invention will be described. Fig. 1 is a schematic diagram of a liquid injection cap 1 according to a first embodiment of the present invention, Fig. 2 is a schematic diagram illustrating a liquid injection operation using the liquid injection cap 1 of Fig. 1, and Fig. 3 is a schematic diagram of a modified example of the liquid injection cap 1. In this embodiment, the liquid L is a culture medium, the liquid injection container 5 is a liquid injection bottle that contains the culture medium, the storage container 6 is a cell culture flask, and the liquid injection operation will be described as being performed in a clean room. In the drawings in this application, the size of each component is exaggerated as appropriate for ease of explanation, and the illustrated components do not represent their actual size.

[0021] As shown in FIGS. 1 and 2 , a liquid injection cap 1 according to a first embodiment of the present invention includes a lid 2 that can be detachably attached to a neck 52 of a liquid injection container 5 that contains a liquid L (culture medium), a liquid injection tube 3 that extends upward and communicates with a first through-hole 21 provided in a top plate 20 of the lid 2, and an air intake tube 4 that extends upward and communicates with a second through-hole 22 provided in the top plate 20. The air intake tube 4 includes a main body 43 that extends upward from the second through-hole 22 and an air intake hole 42 provided at the tip of the main body 43. The inner diameter of the air intake hole 42 is smaller than the inner diameter of the main body 43, thereby forming a step 41 in the flow path of the air intake tube 4. In other words, the step 41 refers to the portion where the inner diameter of the air intake tube 4 changes abruptly in the longitudinal direction.

[0022] The tip of the injection tube 3 is cut at an angle relative to the vertical axis so that the side that tilts the injection container 5 is longer, minimizing dripping. Furthermore, cutting the tip at an angle minimizes problems such as the formation of a film at the tip after injection. The top plate 20 is disk-shaped and can cover the opening of the injection container 5. It has a cylindrical skirt wall 23 hanging down from its periphery. An internal thread is provided on the inner peripheral surface of the cylindrical skirt wall 23. The injection container 5 includes a cylindrical body 51 with a bottom that contains the liquid L and a neck 52 with an outer diameter smaller than the outer diameter of the body 51. An opening is provided at the tip of the neck 52. An external thread (not shown) on the outer peripheral surface of the neck 52 can be threadedly engaged with the internal thread of the cylindrical skirt wall 23, allowing the injection cap 1 to be screwed onto the injection container 5 and sealed.

[0023] The length of the liquid injection tube 3 from the lid 2 is not particularly limited, but is preferably 5 to 100 mm, more preferably 10 to 80 mm, and even more preferably 20 to 50 mm. The inner diameter is preferably 1 to 10 mm, and even more preferably 3 to 5 mm. The flow rate per unit time of the liquid injection tube 3 is 1.0 ml / s to 20 ml / s, preferably 2.0 ml / s to 15 ml / s, and even more preferably 2.5 ml / s to 10.3 ml / s. The combination of the inner diameter and flow rate of the liquid injection tube 3 can be set so that when the inner diameter is in the range of 3 mm to 5 mm, the flow rate is in the range of 2.0 ml / s to 10.0 ml / s. Furthermore, assuming that the flow rate is proportional to the cross-sectional area of ​​the inner diameter, the flow rate can be set to be in the range of approximately 15 ml / s to 40 ml / s when the inner diameter is in the range of 6 mm to 10 mm. The length, inner diameter, and flow rate of the liquid injection tube 3 can be freely selected and combined as long as the flow rate per unit time is constant.

[0024] The length of the intake tube 4 extending from the lid 2 is not particularly limited, but is preferably 5 to 100 mm, more preferably 10 to 50 mm, and even more preferably 10 to 20 mm. The inner diameter of the intake tube 4 (main body 43) is preferably 2.0 mm to 10.0 mm, more preferably 2.0 mm to 7.0 mm, and even more preferably 2.0 mm to 4.0 mm. The inner diameter of the intake hole 42 is preferably 0.5 mm to 3.0 mm, more preferably 1.0 to 2.0 mm, even more preferably 1.1 mm to 1.6 mm, and most preferably 1.2 mm to 1.4 mm. The lid 2, injection tube 3, and intake tube 4 can be integrally molded by injection molding or the like. In the case of integral molding, the inner diameters of the main body 43 and the intake hole 42 can be gently tapered to form a draft angle.

[0025] As shown in FIG. 2A , when using the liquid injection cap 1, the liquid injection cap 1 is attached to a liquid injection container 5 containing liquid L and tilted downward, and liquid is injected into the storage container 6. Tilting the liquid injection container 5 downward means that the liquid injection container 5, which is normally placed vertically on a workbench, is tilted horizontally and then further tilted (i.e., rotated) so that the liquid injection cap 1 is positioned below the liquid injection container 5. When tilted downward in this manner, the base ends of both the liquid injection tube 3 and the air intake tube 4 are covered with liquid L. However, as shown in FIG. 2A , tilting the liquid injection tube 3 so that it is lower than the air intake tube 4 prompts the liquid L to be discharged from the liquid injection tube 3 first, and then prompts air intake from the air intake tube 4 to allow for the discharge of the liquid L. This allows air to be constantly suctioned into the air intake tube 4 during the liquid injection operation, minimizing problems such as leakage of the liquid L from the air intake hole 42.

[0026] The storage container 6 into which the liquid L is poured can be arranged in various ways. For example, when the storage container 6 is a cell culture flask, it is preferably arranged so that the main surfaces 61 and 62 are parallel to the rotation axis, as shown in FIG. 2A. It is also preferable to place the storage container 6 on an inclined base S or the like so that the liquid L discharged from the liquid pouring tube 3 flows gently along the inclined surface of the storage container 6. This minimizes bubbling of the culture medium, which may inhibit cell culture. Furthermore, as shown in FIG. 2A, by arranging the main surfaces 61 (culture surface) of the main surfaces 61 and 62 so that the main surface 61 (culture surface) is above the main surface 62, it is possible to minimize the problem of the liquid L directly hitting the main surface 61 (culture surface), which inhibits cell culture.

[0027] As shown in FIG. 2B, an air A reservoir (space) with a certain volume is formed inside the intake tube 4 (main body 43), and the buoyancy of this air A reservoir repels the pressure of the liquid L. Furthermore, because the tip of the intake tube 4 is covered with a lid having a small intake hole 42, the air A has no escape route and can continue to resist the pressure of the liquid L. During injection, outside air constantly enters through the intake hole 42, pushing out the air A and forming continuous bubbles, allowing the air A to continue to resist the pressure of the liquid L. In this way, by providing a step 41 within the flow path of the intake tube 4 and configuring an air reservoir space with a large inner diameter (inside the main body 43) and a portion with a small inner diameter that restricts the intake amount (intake hole 42), the possibility of the liquid L entering the intake tube 4 can be minimized while maintaining a constant intake amount per unit time.

[0028] As a modified example of the liquid injection cap 1, as shown in Fig. 3, the portion of the intake tube 4 where the intake hole 42 is located can be extended upward, thereby further increasing the distance between the intake hole 42 and the cover 2. As a result, even if the liquid L enters the intake tube 4, it takes time for the liquid L to reach the intake hole 42, and by performing a further injection operation during this time, outside air can be taken into the intake tube 4 and the entered liquid L can be pushed back. The inner diameter of the intake hole 42 (upper side) can be configured to be the same as or smaller than the inner diameter of the intake hole 42 (lower side).

[0029] The amount of air flowing into the injection container 5 as the liquid L is discharged from the injection tube 3 is limited by the small-diameter air intake hole 42 and remains constant. Furthermore, the air A becomes continuous bubbles within the injection container 5, and when the amount of air intake per unit time becomes constant, the amount of liquid injected per unit time also becomes constant. In this case, since the injection time and the amount of liquid injected are proportional to each other during the injection operation, the relationship between the amount of liquid injected and the time required for injection can be measured in advance, and the amount of liquid injected can be determined from the injection time based on the measurement results, allowing for highly accurate injection operations. To maintain continuous bubbles in the injection container 5 for a longer period of time, the air intake tube 4 can be configured so that it does not extend below the lid 2, i.e., into the injection container 5.

[0030] When the amount of liquid to be poured into the storage container 6 reaches a predetermined amount, the tilt of the liquid pouring container 5 is released (for example, the liquid pouring container 5 is horizontally tilted, then tilted further upward, and then returned to a vertical position (i.e., reverse rotation)), and the liquid L is moved to the opposite side of the lid 2, thereby completing the pouring of the liquid L. By repeatedly tilting the liquid pouring container 5 downward and then releasing the tilt, multiple pouring operations can be performed continuously without interruption. If the inner diameter of the base end side (second through-hole 22) of the intake tube 4 is small, the buoyancy of the air A cannot resist the pressure of the liquid L when the liquid L is moved from the lid 2, and the liquid L covering the base end side (second through-hole 22) of the intake tube 4 forms a film, which can cause clogging. However, the intake tube 4 (main body 43) of the present invention has a relatively large inner diameter (i.e., a large opening area), so that a film is less likely to form, and clogging can be minimized.

[0031] The infusion container 5 can also be tilted by rotating it around a predetermined axis using a robot. For example, by setting the predetermined axis at the tip of the infusion tube 3 attached to the infusion container 5 and using a rotating device or the like to rotate (tilt) it around the axis, continuous infusion operations can be performed automatically. Furthermore, by measuring the desired amount of liquid to be injected into the storage container 6 and the injection time required for the injection in advance, storing the measured injection time in the robot's memory, and repeating the tilting and releasing of the infusion container 5 using a program, highly accurate infusion operations can be automated. Furthermore, by associating the predetermined axis with the TCP and aligning the rotation axis of the sixth axis of a six-axis articulated robot with the rotation axis of the TCP, the rotation (tilting) of the infusion container 5 as described above can be performed simply by rotating the sixth axis.

[0032] In the past, for example, when injecting 75 ml of culture medium into a culture flask, it was necessary to repeatedly perform the cumbersome task of inserting a pipette into the injection bottle, aspirating the culture medium, visually checking the amount of 75 ml, and then moving the pipette to inject the medium into the culture flask. By using the present invention, for example, an injection cap 1 can be attached to an injection container 5 containing approximately 480 ml of culture medium, and the injection operation can be performed for the pre-measured time required to inject 75 ml. This can be performed continuously (six times) while replacing six separate storage containers 6. This replaces the cumbersome task of the past with the simple task of repeatedly tilting the injection container 5. This eliminates the need to operate and move the pipette, visually check the amount, and significantly reduces the time required for the injection operation.

[0033] Furthermore, when using a conventional pipette, the suction and discharge of culture medium were performed through a single flow path, which could result in dripping due to operational errors. However, by using the present invention, the suction of culture medium is no longer necessary, and the discharge of culture medium and the intake of air are performed through separate flow paths, minimizing dripping. Furthermore, by using the present invention, the injection of culture medium is performed through a tube, so the injection direction and injection range of culture medium are limited by the protruding direction and diameter of the tube. This improves the accuracy of injection aiming, allowing the injection to be performed accurately with respect to a storage container 6 with a small opening, such as the neck 63 of a cell culture flask, minimizing dripping.

[0034] As described above, according to the present invention, it is possible to prevent the amount of liquid injected from becoming unstable, and to perform efficient, rapid, and highly accurate continuous liquid injection work. Furthermore, since the device has a simple structure that does not use a check valve, it can be manufactured inexpensively. Furthermore, according to the present invention, the liquid injection operation can be performed by accurately aiming at the storage container, and the occurrence of liquid dripping can be minimized, making it suitable for producing cell cultures in clean rooms, etc.

[0035] [Second embodiment] Next, a second embodiment of the present invention will be described. FIG. 4 is a schematic diagram of a liquid injection cap 1′ according to a second embodiment of the present invention, FIG. 5 is a schematic diagram illustrating the liquid injection operation using the liquid injection cap 1′ of FIG. 4, and FIG. 6 is a schematic diagram illustrating the liquid injection principle of FIG. 5.

[0036] As shown in FIG. 4 , a liquid injection cap 1′ according to a second embodiment of the present invention includes a lid 2, a liquid injection tube 3 extending upward and communicating with a first through-hole 21 provided in a top plate 20 of the lid 2, and an air intake tube 4 extending upward and communicating with a second through-hole 22 provided in the top plate 20. The air intake tube 4 includes a main body 43 extending upward from the second through-hole 22 and an air intake hole 42 provided at the tip of the main body 43. The inner diameter of the air intake hole 42 is smaller than the inner diameter of the main body 43, thereby forming a step 41 in the flow path of the air intake tube 4. In this embodiment, the liquid injection cap 1′ further includes an extension tube 44 extending downward and communicating with the second through-hole 22 ( FIG. 4B ). That is, the air intake tube 4 extends downward from the lid via the extension tube 44.

[0037] The length of the extension tube 44 extending from the lid 2 is not particularly limited, but is preferably 5 to 100 mm, more preferably 10 to 50 mm, and even more preferably 15 to 30 mm. The inner diameter of the extension tube 44 is preferably 2.0 mm to 10.0 mm, more preferably 2.0 mm to 7.0 mm, and even more preferably 2.0 mm to 4.0 mm. The extension tube 44 can be molded integrally with the main body 43 by injection molding or the like. Alternatively, an annular groove may be formed on the inner surface of the lid 2 so as to surround the second through-hole 22, and the intake tube 4 may be extended downward by inserting the extension tube 44 into the annular groove.

[0038] FIG. 5A shows a liquid injection cap 1 according to the first embodiment, and FIG. 5B shows a liquid injection cap 1' according to the second embodiment, both attached to a liquid injection container 5 containing liquid L and tilted (rotated) at the same angle. As shown in FIG. 5A, tilting the liquid injection tube 3 so that it is lower than the air intake tube 4 promotes the discharge of liquid L from the liquid injection tube 3 first, and promotes the intake of air from the air intake tube 4 to discharge the liquid L. This causes air to be constantly sucked into the air intake tube 4 during the liquid injection operation, minimizing problems such as leakage of liquid L from the air intake hole 42.

[0039] That is, in FIG. 5A, the liquid L in the liquid injection container 5 first reaches the base end (first through-hole 21) of the liquid injection tube 3, and when the tube is tilted further, the liquid level L reaches the base end (second through-hole 22) of the air intake tube 4. On the other hand, as shown in FIG. 5B, when the air intake tube 4 is extended (stretched) downward, the liquid L does not reach the base end of the air intake tube 4 (the base end of the extension tube 44) when tilted at the same angle. That is, by extending the air intake tube 4 downward (toward the liquid injection container 5), the time it takes for the liquid L to reach the air intake tube 4 can be delayed. As a result, the liquid L reaches the air intake tube 4 when the discharge speed of the liquid L from the liquid injection tube 3 (the intake speed from the air intake tube 4) has increased to a certain extent, and therefore clogging of the air intake tube 4 can be more reliably prevented.

[0040] FIG. 6 is a schematic diagram illustrating the principle of injection shown in FIG. 5. By applying Bernoulli's theorem, the discharge speed (flow rate) v from the injection tube 3 can be calculated using the difference in height h (i.e., the streamline) between the tip of the tube 3 and the base end of the intake tube 4. The flow rate v1 (FIG. 5A) when the injection cap 1 according to the first embodiment is used and the flow rate v2 (FIG. 5B) when the injection cap 1' according to the second embodiment is used are given by the following equations:

number

[0041] Here, as is clear from FIGS. 5A and 5B, since h'<h'', v1<v2. That is, when the liquid injection cap 1' is used, the flow rate becomes faster, so the clogging of the intake tube 4 can be more reliably suppressed. On the other hand, even when the flow rate becomes slower when using the liquid injection cap 1, a certain amount of liquid L can be slowly injected. Therefore, when using a small-volume culture flask or the like, it is advantageous that the liquid L is not overfilled. Since the flow rate is constant until the liquid level of the liquid L reaches the intake tube 4 (until continuous bubbles continue to be generated) during liquid injection, the length of the extension tube 44 can be set shorter than, for example, the length of the neck portion 52 of the liquid injection container 5, so that a constant flow rate can be maintained for a longer time.

[0042] As described above, according to the present invention, it is possible to prevent the instability of the liquid injection amount, and efficient, rapid, and highly accurate continuous liquid injection operations are possible. In addition, since it has a simple structure without using a check valve, it can be manufactured at a low cost. Further, according to the present invention, the liquid injection operation can be performed by accurately aiming at the storage container, and the occurrence of liquid dripping can be minimized. Therefore, it is suitable for the production of cell cultures in a clean room or the like. Furthermore, by increasing the liquid injection speed, the clogging of the intake tube can be more reliably suppressed.

[0043] Although the liquid injection cap according to the present invention has been described, the present invention is not limited thereto. In the present invention, each component can be replaced with any component that can exhibit the same function, or any component can be added.

[0044] Example 1 The liquid injection cap 1 was attached to the liquid injection container 5, and the liquid injection operation was performed. The inner diameter of the intake tube 4 (main body 43) was set to 3.0 mm, and the inner diameter of the intake hole 42 of the liquid injection cap 1 was varied (0.8 mm to 1.7 mm). The liquid injection operation was performed when a culture medium was used as the liquid L and when water was used as the liquid L, and the occurrence of leakage from the intake hole 42 of the intake tube 4, i.e., clogging of the intake tube 4, was confirmed. As shown in Table 1, when a culture medium was used, no leakage occurred when the inner diameter of the intake hole 42 was 1.2 mm to 1.4 mm. Furthermore, when water was used, no leakage occurred when the inner diameter of the intake hole 42 was 1.1 mm to 1.6 mm. [Table 1] [Explanation of symbols]

[0045] 1 Filling cap 2 Lid 21 First through hole 22 Second through hole 23 Cylindrical skirt wall 3 Infusion tube 4 intake tube 41 Step 42 Air intake 43 Main body 44 Extension tube 5. Injection container 51 Main body 52 Neck 6. Storage container 61 Main surface (culture surface) 62 Main Surface 63 Neck L liquid A. Air S base

Claims

1. A liquid injection cap to be attached to a liquid injection container includes a lid body that can be detachably attached to the liquid injection container, and a liquid injection tube and an air intake tube that extend upward from the lid body, wherein a step portion is provided in the flow path of the air intake tube and no check valve is provided, and the step portion includes a portion where the inner diameter of the air intake hole of the air intake tube is smaller than the inner diameter of the air intake tube, so that the inner diameters of the air intake tube and the air intake hole do not change during liquid injection. The liquid injection cap.

2. 2. The liquid filling cap according to claim 1, wherein the inner diameter of the intake hole of the intake tube is 0.5 mm to 3.0 mm.

3. 3. The liquid filling cap according to claim 1, wherein the inner diameter of the intake tube is 2.0 mm to 10.0 mm.

4. 4. The liquid filling cap according to claim 1, wherein the length of the intake tube extending upward from the lid body is 5 to 100 mm.

5. 5. The liquid filling cap according to claim 1, further comprising an extension tube that extends the intake tube downward from the lid body.

6. 6. The liquid filling cap according to claim 5, wherein the extension tube has an inner diameter of 2.0 mm to 10.0 mm.

7. 7. The liquid filling cap according to claim 5, wherein the extension tube has a length of 5 to 100 mm.

8. 8. The liquid filling cap according to claim 5, wherein the length of the extension tube is shorter than the length of the neck of the liquid filling container.

9. A method for injecting a liquid, the method comprising: (a) the step of preparing an injection container containing a liquid; (b) the step of attaching an injection cap to the injection container, the injection cap including a lid that can be removably attached to the injection container, an injection tube and an intake tube extending upward from the lid, a stepped portion provided in the flow path of the intake tube and not provided with a check valve, the stepped portion including a portion where the inner diameter of the intake hole of the intake tube is smaller than the inner diameter of the intake tube, and configured so that the inner diameters of the intake tube and intake hole do not change during injection; (c) the step of tilting the injection container downward and injecting the liquid through the injection tube; and (d) the step of releasing the downward tilt of the injection container to terminate the injection.

10. The method according to claim 9, wherein the infusion container is tilted by rotating the infusion container around a predetermined axis.

11. 11. The method according to claim 9 or 10, further comprising a step (e) of repeating steps (c) and (d) to perform a plurality of injection operations successively.

12. The method according to any one of claims 9 to 11, further comprising the step of: (f) determining the amount of liquid injected from the injection time.

13. The method according to any one of claims 9 to 12, wherein the filling cap further comprises an extension tube that extends the intake tube downward from the lid body.

14. A method according to any one of claims 9 to 13, wherein step (c) includes tilting the infusion tube so that it is lower than the intake tube.

15. A method as described in any one of claims 9 to 14, wherein step (c) includes tilting the intake tube so that an air pocket is formed within the intake tube.

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