WO2021223150A1 - 用以调节液体分布的容器及其方法 - Google Patents

用以调节液体分布的容器及其方法 Download PDF

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Publication number
WO2021223150A1
WO2021223150A1 PCT/CN2020/088960 CN2020088960W WO2021223150A1 WO 2021223150 A1 WO2021223150 A1 WO 2021223150A1 CN 2020088960 W CN2020088960 W CN 2020088960W WO 2021223150 A1 WO2021223150 A1 WO 2021223150A1
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WIPO (PCT)
Prior art keywords
chamber
connecting portion
sub
container
liquid
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PCT/CN2020/088960
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English (en)
French (fr)
Inventor
张浩祯
刘惟凡
陈弘叡
冯爵敏
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元锦生物科技股份有限公司
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Priority to PCT/CN2020/088960 priority Critical patent/WO2021223150A1/zh
Publication of WO2021223150A1 publication Critical patent/WO2021223150A1/zh

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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B05—SPRAYING OR ATOMISING IN GENERAL; APPLYING FLUENT MATERIALS TO SURFACES, IN GENERAL
    • B05C—APPARATUS FOR APPLYING FLUENT MATERIALS TO SURFACES, IN GENERAL
    • B05C17/00—Hand tools or apparatus using hand held tools, for applying liquids or other fluent materials to, for spreading applied liquids or other fluent materials on, or for partially removing applied liquids or other fluent materials from, surfaces
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M3/00—Tissue, human, animal or plant cell, or virus culture apparatus

Definitions

  • the present invention relates to a container for storing fluid.
  • it refers to a container for adjusting the distribution of liquid and a method for adjusting the distribution of liquid in the container.
  • the traditional cell culture method is mainly to grow in a Multi-Well Microplate under sterile, proper temperature and nutrient conditions by static culture. After about a period of time, each hole The cells in the cell will grow into a colony (Colony) to achieve the purpose of cell proliferation.
  • the porous microdisk In order to ensure that the cells grow in the best environment, the porous microdisk is usually immersed in the culture medium to prevent the culture medium in the pores from being depleted due to cell growth and reproduction, which may cause cell death.
  • the culture medium may be disturbed during the cell extraction process, causing the cells in the holes to detach and cause contamination, which affects the subsequent Experimental results.
  • the culture medium is directly removed, the cells in the holes may be disturbed, and the cells will even be taken away from the holes with the culture medium.
  • the experimental results will be incorrect and the accuracy will be reduced. Sex, even worse, will lead to the failure of the experiment, so it is indeed necessary to improve.
  • the main objective of the present invention is to provide a container for adjusting the distribution of liquid. It only remains in the sub-chamber, and can be further applied to cell culture and other related fields.
  • the container provided by the present invention for adjusting liquid distribution includes: a first body; a sub-chamber recessed on one side of the first body; and a flexible second
  • the main body can be detachably covered on the first body to close the sub-chamber; an accommodating space is formed between the first body and the second body to enable the accommodating space and the sub-chamber to be able to communicate with each other In communication with each other to accommodate the external liquid; wherein, when one end of the second body is driven by an external force, it is lifted from the first end surface of the first body, and gradually moves away from the second body toward the other end of the second body.
  • the liquid in the containing space is taken away as the second body moves away , And make the liquid in the sub-chamber still remain in the sub-chamber.
  • the container of the present invention further includes a recessed chamber, which is recessed on one side of the second body corresponding to the position of the sub-chamber, and the receiving space is formed by the recessed chamber.
  • the container of the present invention further includes a recessed chamber recessed on one side of the first body, and the sub-chamber is recessed on the bottom surface of the recessed chamber and communicates with the recess formed by the recessed chamber. Accommodate space.
  • the container of the present invention further includes a first connecting portion and a second connecting portion that can be separated or combined with each other, which are separately provided on the end surfaces between the first body and the second body facing each other When the first body is attached to the second body, the first connecting portion and the second connecting portion are combined with each other, so that the second body and the first body can be connected to each other.
  • first connecting portion and the second connecting portion are respectively an annular ring groove and an annular convex wall that can be complementarily fitted with each other.
  • the first body further includes a residual liquid storage chamber, which is recessed on one side of the first body apart from the sub-chamber.
  • the container of the present invention further includes: a third connecting portion, which is provided on a side of the first body facing the second body, and is located on the ring groove on the first body At a position outside the surrounding range; a fourth connecting portion, which is separable or combined with the third connecting portion, is provided on one side of the second body, and is located on the convex wall on the second body When the first body is attached to the second body, the third connecting portion and the fourth connecting portion are combined with each other, so that the second body and the The first bodies are connected to each other.
  • the container of the present invention further includes a third body.
  • the third body When the third body is connected to the first body, the second body is sandwiched between the first body and the third body. between.
  • the container further includes a fifth connecting portion and a sixth connecting portion that can be separated or combined with each other, which are separately provided on the third body and the first body, and when the third body is attached to the first body At the same time, the fifth connecting portion and the sixth connecting portion are combined with each other, and the third body is combined with the first body.
  • a method for adjusting liquid distribution uses the container for adjusting liquid distribution disclosed in the present invention to fill the containing space and the sub-chamber with liquid, and apply an external force to make One end of the second body separates from the first body and gradually moves away from the first body toward the other end of the second body.
  • the viscosity is generated by the contact between the liquid surface and the second body
  • the force and the surface tension of the liquid itself cause the liquid in the containing space to be taken away with the distance of the second body, and make the liquid in the sub-chamber remain in the sub-chamber.
  • FIG. 1 is a three-dimensional assembly diagram of a container for adjusting liquid distribution disclosed in the first embodiment of the present invention.
  • Fig. 2 is a perspective exploded view of Fig. 1 according to the first embodiment of the present invention.
  • Fig. 3 is a cross-sectional view taken along line 3-3 of Fig. 1 according to the first embodiment of the present invention.
  • Fig. 4 is an action diagram of the first embodiment of the present invention following Fig. 3, which shows the process of removing the second body from the first body.
  • FIG. 5A is a perspective view of the first embodiment of the present invention continued from FIG. 4, which shows the state that the second body has been removed from the first body.
  • Fig. 5B is a cross-sectional view taken along line 5-5 of Fig. 5A according to the first embodiment of the present invention.
  • Fig. 6A is a partial photomicrograph of the first group test of Test Example 1 of the first embodiment of the present invention, which shows the number of beads in each observation area when the second body has not been removed.
  • Fig. 6B is a partial photomicrograph of the first set of experiments of Test Example 1 of the first embodiment of the present invention, which shows the number of beads in each observation area after the second body is removed.
  • Fig. 7A is a partial photomicrograph of the second group test of Test Example 1 of the first embodiment of the present invention, which shows the number of beads in each observation area when the second body has not been removed.
  • FIG. 7B is a partial photomicrograph of the second group of experiments of Experimental Example 1 of the first embodiment of the present invention, which shows the number of microbeads in each observation area after the second body is removed.
  • Fig. 8A is a partial photomicrograph of the third group test of Test Example 1 of the first embodiment of the present invention, which shows the number of beads in each observation area when the second body has not been removed.
  • Fig. 8B is a partial photomicrograph of the third group test of Test Example 1 of the first embodiment of the present invention, which shows the number of beads in each observation area after the second body is removed.
  • Fig. 9A is a partial photomicrograph of the first group test of Test Example 2 of the first embodiment of the present invention, which shows the number of beads in each observation area when the second body has not been removed.
  • Fig. 9B is a partial photomicrograph of the first group test of Test Example 2 of the first embodiment of the present invention, which shows the number of beads in each observation area after the second body is removed.
  • Fig. 10A is a partial photomicrograph of the second group test of Test Example 2 of the first embodiment of the present invention, which shows the number of beads in each observation area when the second body has not been removed.
  • Fig. 10B is a partial photomicrograph of the second group of experiments of Experimental Example 2 of the first embodiment of the present invention, which shows the number of microbeads in each observation area after the second body is removed.
  • FIG. 11A is a partial photomicrograph of the third group test of Test Example 2 of the first embodiment of the present invention, which shows the number of beads in each observation area when the second body has not been removed.
  • Fig. 11B is a partial photomicrograph of the third group of test of Test Example 2 of the first embodiment of the present invention, which shows the number of beads in each observation area after the second body is removed.
  • Fig. 12A is a partial photomicrograph of the first group test of Test Example 3 of the first embodiment of the present invention, which shows the number of cells in each observation area when the second body has not been removed.
  • Fig. 12B is a partial photomicrograph of the first group test of Test Example 3 of the first embodiment of the present invention, which shows the number of cells in each observation area after the second body is removed.
  • Fig. 12C is a partial photomicrograph of the first group experiment of Test Example 3 of the first embodiment of the present invention, which shows the number of cells in each observation area after the cells in the upper right observation area in the container are sucked out.
  • Fig. 13A is a partial photomicrograph of the second group test of Test Example 3 of the first embodiment of the present invention, which shows the number of cells in each observation area when the second body has not been removed.
  • Fig. 13B is a partial photomicrograph of the second group test of Test Example 3 of the first embodiment of the present invention, which shows the number of cells in each observation area after the second body is removed.
  • Fig. 13C is a partial photomicrograph of the second group of test of Test Example 3 of the first embodiment of the present invention, which shows the number of cells in each observation area after the cells in the upper right observation area in the container are sucked out.
  • Fig. 14A is a partial photomicrograph of the third group test of Test Example 3 of the first embodiment of the present invention, which shows the number of cells in each observation area when the second body has not been removed.
  • Fig. 14B is a partial photomicrograph of the third group test of Test Example 3 of the first embodiment of the present invention, which shows the number of cells in each observation area after the second body is removed.
  • 14C is a partial photomicrograph of the third group test of Test Example 3 of the first embodiment of the present invention, which shows the number of cells in each observation area after the cells in the upper right observation area in the container are sucked out.
  • Fig. 15A is a partial photomicrograph of the first group of test of Test Example 3 of the first embodiment of the present invention, which shows the number of cells in each observation area when the second body has not been removed.
  • Fig. 15B is a partial photomicrograph of the first group test of Test Example 3 of the first embodiment of the present invention, which shows the number of cells in each observation area after the second body is removed.
  • 15C is a partial photomicrograph of the first set of experiments of Test Example 3 of the first embodiment of the present invention, which shows the number of cells in each observation area after the cells in the upper right observation area in the container are sucked out.
  • Fig. 16A is a partial photomicrograph of the second group test of Test Example 3 of the first embodiment of the present invention, which shows the number of cells in each observation area when the second body has not been removed.
  • Fig. 16B is a partial photomicrograph of the second group test of Test Example 3 of the first embodiment of the present invention, which shows the number of cells in each observation area after the second body is removed.
  • Fig. 16C is a partial photomicrograph of the second group test of Test Example 3 of the first embodiment of the present invention, which shows the number of cells in each observation area after the cells in the upper right observation area in the container are sucked out.
  • Fig. 17A is a partial photomicrograph of the third group test of Test Example 3 of the first embodiment of the present invention, which shows the number of cells in each observation area when the second body has not been removed.
  • Fig. 17B is a partial photomicrograph of the third group test of Test Example 3 of the first embodiment of the present invention, which shows the number of cells in each observation area after the second body is removed.
  • Fig. 17C is a partial photomicrograph of the third group test of Test Example 3 of the first embodiment of the present invention, which shows the number of cells in each observation area after the cells in the upper right observation area in the container are sucked out.
  • FIG. 18 is a three-dimensional assembly view of a container for adjusting liquid distribution disclosed in the second embodiment of the present invention.
  • FIG. 19 is a perspective exploded view of FIG. 18 according to the second embodiment of the present invention.
  • Fig. 20 is a cross-sectional view taken along line 20-20 of Fig. 18 according to the second embodiment of the present invention.
  • FIG. 21 is an action diagram of the second embodiment of the present invention continued from FIG. 20, which shows the process of removing the second body from the first body.
  • FIG. 22 is a perspective view of the second embodiment of the present invention continued from FIG. 21, which shows the state that the second body has been removed from the first body.
  • FIG. 23 is a cross-sectional view of the second embodiment of the present invention taken along the line 23-23 in FIG. 22.
  • FIG. 23 is a cross-sectional view of the second embodiment of the present invention taken along the line 23-23 in FIG. 22.
  • FIG. 24 is a three-dimensional assembly view of a container for adjusting liquid distribution disclosed in the third embodiment of the present invention.
  • FIG. 25 is a perspective exploded view of FIG. 24 according to the third embodiment of the present invention.
  • Fig. 26 is a cross-sectional view taken along line 25-25 of Fig. 25 according to the third embodiment of the present invention.
  • 10, 10A, 10B first body; 11: first body; 12, 12A: first end face; 13: ring wall; 14: first connection part; 141, 141A: ring groove; 15: third connection 151, 151A: recess; 16: residual liquid storage chamber; 17: fifth connecting part; 20, 20A: recess; 30, 30A: sub-chamber; 40, 40A, 40B: second body; 41: second Body; 42, 42A: second end surface; 43: second connecting part; 431, 431A: convex wall; 44: fourth connecting part; 441, 441A: convex part; 45: recessed part; 451: clearance; 50 , 50A: containing space; 60: liquid; 70: third body; 71: sixth connecting part; F1: viscosity; F2: surface tension; W: gravity; 80: cells.
  • the container for adjusting the distribution of liquid 60 disclosed in the first embodiment of the present invention includes a first body 10, a recess 20, a sub-chamber 30, and a second body 40 And an accommodation space 50.
  • the first body 10 has a first body 11, which is in the shape of a flat plate, and a first end surface 12 located on one side of the first body 11.
  • the recessed chamber 20 is recessed on one side of the first body 10, and the first body 11 is formed with a ring 13 surrounding the peripheral side of the recessed chamber 20.
  • the recessed chamber 20 is recessed on the first end surface 12 of the first body 10, and the shape of the recessed chamber 20 may be, but not limited to, an elliptical shape or an elongated shape.
  • the accommodating space 50 is formed around the wall of the recessed chamber 20.
  • the sub-chamber 30 is recessed on one side of the first body 10.
  • the sub-chamber 30 is recessed on the bottom surface of the recess 20, wherein the number of the sub-chambers 30 can be configured according to user requirements, and in this example, the number of the sub-chambers 30 is two groups The number of 6*5 arrays.
  • the second body 40 is made of flexible material and has a second body 41, which is detachably covered on the first end surface 12 of the first body 10, and at the same time, is also covered on the top side of the ring wall 13
  • the recess 20 is closed; and a second end surface 42 is opposite to the first end surface 12 and is located on one side of the second body 41.
  • the container further includes a first connecting portion 14 and a second connecting portion 43 that can be separated or combined with each other.
  • first connecting portion 14 and the second connecting portion 43 are combined with each other, so that the second body 40 and the first body 10 can be connected to each other.
  • the first connecting portion 14 and the second connecting portion 43 are respectively a ring-shaped ring groove 141 and a ring-shaped protrusion 431 that can be complementary fitted to each other, and the concave chamber 20
  • the sub-chamber 30 and the sub-chamber 30 are both located in the range surrounded by the ring groove 141 on the first body 11.
  • the protruding wall 431 and the ring groove 141 are joined, thereby preventing the space formed by the recessed chamber 20 and the sub-chamber 30 from communicating with the outside world .
  • the ring groove 141 is separated from the sub-chamber 30 without communicating with each other as a residual liquid storage space.
  • the remaining liquid storage space is different from the ring groove 141, and is separately provided on the first body 11, and is not connected to the recessed chamber 20 and the sub-chamber 30.
  • the container of the present invention further includes: a third connecting portion 15 provided on the side of the first body 10 opposite to the second body 40 and located on the ring groove 141 on the first body 10
  • a fourth connecting portion 44 and the third connecting portion 15 are detachably or combinedly provided on one side of the second body 40, and are located on the convex wall 431 in the first
  • the second body 40 is located at a position outside the surrounding range; wherein, when the first body 10 is attached to the second body 40, the third connecting portion 15 and the fourth connecting portion 44 are combined with each other.
  • the second body 40 and the first body 10 are connected to each other.
  • the third connecting portion 15 and the fourth connecting portion 44 are respectively a concave portion 151 and a convex portion 441 that can be complementarily fitted with each other.
  • the concave portion 151 may be, but not limited to, a straight, T-shaped or cross-shaped protrusion structure
  • the convex portion 441 may be, but not limited to, a straight, T-shaped or cross-shaped concave structure.
  • the liquid 60 is filled in the concave chamber 20 and the sub-chamber 30, as shown in FIG. 3.
  • An external force is applied to separate one end of the second body 40 from the first end surface 12 of the first body 10 and gradually move away from the first body 10 toward the other end of the second body 40.
  • the surface tension F2 overcomes the gravity W of the liquid 60, so that the liquid 60 in the recessed chamber 20 is taken away as the second body 40 moves away, and the liquid 60 in the sub-chamber 30 remains in the cavity. In the subroom 30.
  • the surface tension F2 of the liquid 60 is between 56.5 dyne/cm and 88.9 dyne/cm
  • the viscosity of the liquid 60 is between 1.10 cP and 5.62 cP
  • the height of the sub-chamber 30 is equal to that of the alcove 20.
  • the height ratio is between 12:1 and 3:5.
  • the liquid 60 carried away with the second body 40 can also be removed by the first-rate outlet provided on the first body 10.
  • the second body 40 is completely removed from the first body 10, so that part of the liquid 60 remains in the sub-chamber 30, and the ring groove 141 can store the liquid 60 taken away from the concave chamber 20, As shown in Figure 5A and Figure 5B.
  • the present invention can be applied to the harvesting of cultured cells 80.
  • the operation steps are as follows. First, the sub-chamber 30 of the container contains cultured mature cells 80, as shown in the partial enlarged part of FIG. 3; Disclosure of the implementation steps of the method described, so that the sub-chambers 30 originally communicated with each other through the liquid 60 filled with the recessed chamber 20 are changed to a state of being independent and disconnected (as shown in FIG. 5A and FIG. 5B). As shown), at the same time, the cells 80 contained in the sub-chamber 30 are not disturbed, so as to maintain the number and status of the cells 80 contained in the culture.
  • the CHO-K1 cell is stained with CMFDA (CellTracker TM C7025, Invitrogen) (1:20) to form 4x10 ⁇ 4cell/ml, 1ul is placed in the sub-chambers 30, and then the second body 40 is covered. , Sent to 37 degrees Celsius, 5% CO2 incubator for 24hrs.
  • the second body 40 is peeled from the first body 10, the changes in the number of cells 80 in the four predetermined areas in the container are observed, and the above steps are repeated for 3 sets of tests.
  • test results are as follows As shown in Table 2, Figure 12A, Figure 12B, Figure 13A, Figure 13B, Figure 14A, and Figure 14B, it was found that the number of cells 80 did not change significantly, and the slight changes in the number of cells 80 disclosed in Table 2 may be obscured. As a result, the number and state of the cells 80 in the sub-chamber 30 can be effectively maintained after the second body 40 is removed. In addition, as shown in FIG. 12C, FIG. 13C, and FIG. 14C, after the second body 40 is peeled off, the cells 80 in the upper right observation area in the container are sucked out to serve as a positive control group.
  • FIGS. 18 to 20 are schematic diagrams of the second embodiment of the present invention
  • the main difference between the second embodiment and the first embodiment is that the sub-chamber 30A is recessed in the first body 10A.
  • the first end surface 12A, and the recessed chamber 20A is recessed on one side of the second body 40A corresponding to the position of the sub-chamber 30A, which means that the recessed chamber 20A is recessed on the second end surface 42A of the second body 40A,
  • its wall surface surrounds and forms the accommodating space 50A.
  • the first end surface 12A of the first body 10 is further recessed with a residual liquid storage chamber 16, which is not connected to the sub-chamber 30A, but communicates with the ring groove 141A and the recess 151A, and is located in the The ring groove 141A is at a position outside the range surrounded by the first body 10A.
  • the second body 40A further includes a recessed portion 45, which is recessed on the second end surface 42A of the second body 40A adjacent to the edge of the second body 40A.
  • the concave portion 151A is fitted into the convex portion 441A to further fix the connection relationship between the first body 10A and the second body 40A,
  • the recessed portion 45 is not in contact with the first end surface 12A of the first body 10A, so that there is a clearance 451 between the facing end surfaces of the first body portion 11A and the second body portion 41A, and It is connected to the outside, thereby facilitating the user to hold the second body 40A to facilitate subsequent actions.
  • Figures 24 to 26 are schematic diagrams of the third embodiment of the present invention.
  • the container further includes a third body 70, when the third body 70 When connected with the first body 10B, the second body 40B is sandwiched between the first body 10B and the third body 70.
  • the container further includes a fifth connecting portion 17 and a sixth connecting portion 71 that can be separated or combined with each other, which are separately provided on the third body 70 and the first body 10B, when the third body 70 and the When the first body 10B is attached to each other, the fifth connecting portion 17 and the sixth connecting portion 71 are combined with each other, so that the third body 70 is combined with the first body 10B.
  • the fifth connecting portion 17 and the sixth connecting portion 71 are respectively a convex portion and a concave portion that can be complementarily fitted with each other, and a point-to-point ultrasonic wave can be used between the convex portion and the concave portion. Welding is used to connect at least a part of the convex portion to the concave surface of the concave portion, so that the first body 10B and the third body 70 can be temporarily fixed to each other.
  • a tool such as tweezers
  • tweezers can be used to extend into the gap between the convex portion and the concave portion, and then use the The concave surface of the concave portion is the fulcrum, and the end surface of the convex portion is the focus point, and a slight force can be used to break the welding relationship between the convex portion and the concave portion, and the aforementioned joint structure can be easily pried off.

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Abstract

本发明揭露用以调节液体分布的容器包含有一第一本体；一子室，凹设于该第一本体的一侧；一具挠性的第二本体，可拆卸地覆于该第一本体上，以封闭该子室；一容纳空间，被形成于该第一本体与该第二本体之间，并与该子室彼此间能相互连通地容纳外部的液体；当该第二本体的一端受外力驱使而自该第一本体的第一端面上掀开，并朝着该第二本体的另一端的方向逐步地远离该第一本体时，通过该液体表面与该第二本体间接触所产生的黏滞力，与该液体自身的表面张力，使位于该容纳空间中的液体随该第二本体的远离而被带离，并使位于该子室中的液体仍留存于该子室内。

Description

用以调节液体分布的容器及其方法 技术领域
本发明与贮存流体的容器有关，详而言之，指一种用以调节液体分布的容器，以及一种调节容器内液体的分布的方法。
背景技术
传统的细胞培养方式主要为在多孔微盘(Multi-Well Microplate)中，在无菌、适当温度及营养的条件下，以静态培养(Static Culture)方式使其生长，约略一段时间后，每一孔洞中的细胞会长成一个群落(Colony)，以达到细胞增生的目的。
为了确保细胞在最佳的环境中生长，通常会将多孔微盘浸盈于培养液中，避免孔洞内的培养液会因细胞的生长及繁殖而消耗殆尽，以致于造成细胞死亡。
但是，在细胞培养结束后，因多孔微盘仍浸盈于培养液中，在细胞撷取的过程中可能会扰动培养液，使得该些孔洞中的细胞脱离而发生污染的情形，从而影响后续的实验结果。此外，若直接将培养液抽离，也可能会扰乱该些孔洞中的细胞，甚致使细胞会随着培养液而被带离孔洞，如此一来，将会使得实验结果不正确而降低其准确性，更甚者将导致实验失败，故确有必要改良。
发明内容
因此，本发明的主要目的即在提供一种用以调节液体分布的容器，其主要的技术特征为通过第二本体与第一本体分离时，将容器内的部分液体带离，据以使液体仅残留于子室中，并可进一步应用于细胞培养等相关领域。
缘是，为达成上述的目的，本发明所提供用以调节液体分布的容器包含有：一第一本体；一子室，凹设于该第一本体的一侧；一具挠性的第二本体，可拆卸地覆于该第一本体上，可以封闭该子室；一容纳空间，被形成于该第一本体与该第二本体之间，并使该容纳空间与该子室彼此间能相互连通地容纳外部的液体；其中，当该第二本体的一端受外力驱使而自该第一本体的第一端面上掀开，并朝着该第二本体的另一端的方向逐步地远离该第一本体时，通过该液体表面与该第二本体间接触所产生的黏滞力，与该液体自身的表面张力，使位于该容纳空间中的液体随该第二本体的远离而被带离，并使位于该子室中的液体仍留存于该子室内。
在一实施例中，本发明的该容器更包括一凹室，对应该子室的位置而凹设于该第二本体的一侧，并由该凹室形成该容纳空间。
在另一实施例中，本发明的该容器更包括一凹室，凹设于该第一本体的一侧，并且该子室凹设于该凹室底面，并连通该凹室所形成的该容纳空间。
在一实施例中，本发明的该容器更包括有可彼此分离或结合的一第一连接部与一第二连接部，分设于该第一本体与该第二本体之间彼此相向的端面上，当该第一本体与该第二本体相贴合时，该第一连接部与该第二连接部相互结合，可以使该第二本体与该第一本体彼此相接。
其中，该第一连接部为与该第二连接部分别为可彼此互补嵌合的一环状的环凹槽与一环状的凸垣。
其中，该第一本体更包括有一余液储存室，与该子室相间隔地凹设于该第一本体的一侧。
在一实施例中，本发明的该容器更包括更包括：一第三连接部，设于该第一本体相向于该第二本体的一侧，并且位于该环凹槽在该第一本体上所围绕的范围之外的位置上；一第四连接部，与该第三连接部彼此间可分离或结合地设于该第二本体的一侧，并且位于该凸垣在该第二本体上所围绕的范围之外的位置上；其中，当该第一本体与该第二本体相贴合时，该第三连接部与该第四连接部相互结合，可以使令该第二本体与该第一本体彼此相接。
在一实施例中，本发明的该容器更包括一第三本体，当该第三本体与该第一本体相连接时，将该第二本体夹设于该第一本体与该第三本体之间。
其中，该容器更包括有可彼此分离或结合的一第五连接部与一第六连接部，分设于该第三本体与该第一本体上，当该第三本体与该第一本体相贴合时，使该第五连接部与该第六连接部相互结合，令该第三本体结合于该第一本体上。
在本发明的一实施例中揭露一种用以调节液体分布的方法，其使用本发明所揭用以调节液体分布的容器，使液体充盈于该容纳空间与该子室内，并施加一外力使该第二本体的一端自该第一本体上分离，并朝着该第二本体的另一端的方向逐步地远离该第一本体，通过该液体表面与该第二本体间接触所产生的黏滞力，以及该液体自身的表面张力，使位于该容纳空间中的液体随该第二本体的远离而被带离，并使位于该子室中的液体仍留存于该子室内。
附图说明
图1为本发明第一实施例所揭用以调节液体分布的容器的立体组合图。
图2为本发明第一实施例就图1的立体分解图。
图3为本发明第一实施例就图1的3-3剖面图。
图4为本发明第一实施例接续图3的作动图，其显示第二本体在第一本体上卸除的过程。
图5A为本发明第一实施例接续图4的立体示意图，其显示第二本体已自第一本体上卸除的态样。
图5B为本发明第一实施例就图5A的5-5剖面图。
图6A为本发明第一实施例的试验例1的第1组试验的局部显微相片，其显示第二本体尚未卸除时，各个观察区域内的微珠数量。
图6B为本发明第一实施例的试验例1的第1组试验的局部显微相片，其显示第二本体卸除后，各个观察区域内的微珠数量。
图7A为本发明第一实施例的试验例1的第2组试验的局部显微相片，其显示第二本体尚未卸除时，各个观察区域内的微珠数量。
图7B为本发明第一实施例的试验例1的第2组试验的局部显微相片，其显示第二本体卸除后，各个观察区域内的微珠数量。
图8A为本发明第一实施例的试验例1的第3组试验的局部显微相片，其显示第二本体尚未卸除时，各个观察区域内的微珠数量。
图8B为本发明第一实施例的试验例1的第3组试验的局部显微相片，其显示第二本体卸除后，各个观察区域内的微珠数量。
图9A为本发明第一实施例的试验例2的第1组试验的局部显微相片，其显示第二本体尚未卸除时，各个观察区域内的微珠数量。
图9B为本发明第一实施例的试验例2的第1组试验的局部显微相片，其显示第二本体卸除后，各个观察区域内的微珠数量。
图10A为本发明第一实施例的试验例2的第2组试验的局部显微相片，其显示第二本体尚未卸除时，各个观察区域内的微珠数量。
图10B为本发明第一实施例的试验例2的第2组试验的局部显微相片，其显示第二本体卸除后，各个观察区域内的微珠数量。
图11A为本发明第一实施例的试验例2的第3组试验的局部显微相片，其显示第二本体尚未卸除时，各个观察区域内的微珠数量。
图11B为本发明第一实施例的试验例2的第3组试验的局部显微相片，其显示第二本体卸除后，各个观察区域内的微珠数量。
图12A为本发明第一实施例的试验例3的第1组试验的局部显微相片，其显示第二本体尚未卸除时，各个观察区域内的细胞数量。
图12B为本发明第一实施例的试验例3的第1组试验的局部显微相片，其显 示第二本体卸除后，各个观察区域内的细胞数量。
图12C为本发明第一实施例的试验例3的第1组试验的局部显微相片，其显示将容器内右上方观察区域内的细胞吸除后，各个观察区域内的细胞数量。
图13A为本发明第一实施例的试验例3的第2组试验的局部显微相片，其显示第二本体尚未卸除时，各个观察区域内的细胞数量。
图13B为本发明第一实施例的试验例3的第2组试验的局部显微相片，其显示第二本体卸除后，各个观察区域内的细胞数量。
图13C为本发明第一实施例的试验例3的第2组试验的局部显微相片，其显示将容器内右上方观察区域内的细胞吸除后，各个观察区域内的细胞数量。
图14A为本发明第一实施例的试验例3的第3组试验的局部显微相片，其显示第二本体尚未卸除时，各个观察区域内的细胞数量。
图14B为本发明第一实施例的试验例3的第3组试验的局部显微相片，其显示第二本体卸除后，各个观察区域内的细胞数量。
图14C为本发明第一实施例的试验例3的第3组试验的局部显微相片，其显示将容器内右上方观察区域内的细胞吸除后，各个观察区域内的细胞数量。
图15A为本发明第一实施例的试验例3的第1组试验的局部显微相片，其显示第二本体尚未卸除时，各个观察区域内的细胞数量。
图15B为本发明第一实施例的试验例3的第1组试验的局部显微相片，其显示第二本体卸除后，各个观察区域内的细胞数量。
图15C为本发明第一实施例的试验例3的第1组试验的局部显微相片，其显示将容器内右上方观察区域内的细胞吸除后，各个观察区域内的细胞数量。
图16A为本发明第一实施例的试验例3的第2组试验的局部显微相片，其显示第二本体尚未卸除时，各个观察区域内的细胞数量。
图16B为本发明第一实施例的试验例3的第2组试验的局部显微相片，其显示第二本体卸除后，各个观察区域内的细胞数量。
图16C为本发明第一实施例的试验例3的第2组试验的局部显微相片，其显示将容器内右上方观察区域内的细胞吸除后，各个观察区域内的细胞数量。
图17A为本发明第一实施例的试验例3的第3组试验的局部显微相片，其显示第二本体尚未卸除时，各个观察区域内的细胞数量。
图17B为本发明第一实施例的试验例3的第3组试验的局部显微相片，其显示第二本体卸除后，各个观察区域内的细胞数量。
图17C为本发明第一实施例的试验例3的第3组试验的局部显微相片，其显示将容器内右上方观察区域内的细胞吸除后，各个观察区域内的细胞数量。
图18为本发明第二实施例所揭用以调节液体分布的容器的立体组合图。
图19为本发明第二实施例就图18的立体分解图。
图20为本发明第二实施例就图18的20-20剖面图。
图21为本发明第二实施例接续图20的作动图，其显示第二本体在第一本体上卸除的过程。
图22为本发明第二实施例接续图21的立体示意图，其显示第二本体已自第一本体上卸除的态样。
图23为本发明第二实施例就图22的23-23剖面图。
图24为本发明第三实施例所揭用以调节液体分布的容器的立体组合图。
图25为本发明第三实施例就图24的立体分解图。
图26为本发明第三实施例就图25的25-25剖面图。
附图标记
10、10A、10B：第一本体；11：第一身部；12、12A：第一端面；13：环垣；14：第一连接部；141、141A：环凹槽；15：第三连接部；151、151A：凹部；16：余液储存室；17：第五连接部；20、20A：凹室；30、30A：子室；40、40A、40B：第二本体；41：第二身部；42、42A：第二端面；43：第二连接部；431、431A：凸垣；44：第四连接部；441、441A：凸部；45：凹陷部；451：余隙；50、50A：容纳空间；60：液体；70：第三本体；71：第六连接部；F1：黏滞力；F2：表面张力；W：重力；80：细胞。
具体实施方式
请参阅图1至图3，在本发明的第一实施例所揭用以调节液体60分布的容器，其包含一第一本体10、一凹室20、一子室30、一第二本体40及一容纳空间50。
该第一本体10具有一第一身部11，呈平板状；及一第一端面12，位于该第一身部11的一侧。
该凹室20凹设于该第一本体10的一侧，而在该第一身部11上形成有一环绕该凹室20周侧的环垣13。在本实施例中，该凹室20凹设于该第一本体10的第一端面12，且该凹室20的形状可为但不限于呈椭圆形或长条形。此外，该凹室20的壁面围绕形成该容纳空间50。
该子室30凹设于该第一本体10的一侧。在本实施例中，该子室30凹设于该凹室20底面，其中，该子室30的数量可依据使用者需求来配置，而在本例中，该子室30的数量为两组6*5阵列所构成的数量。
该第二本体40具挠性的材质所构成，并具有一第二身部41，可拆卸地覆于该第一本体10的第一端面12上，同时，也覆于该环垣13顶侧而封闭该凹室20；及一第二端面42，与该第一端面12相向并位于该第二身部41的一侧。
此外，该容器更包括有可彼此分离或结合的一第一连接部14与一第二连接部43，分设于该第一本体10与该第二本体40之间彼此相向的端面上，当该第一本体10与该第二本体40相贴合时，该第一连接部14与该第二连接部43相互结合，可以使该第二本体40与该第一本体10彼此相接。在本实施例中，该第一连接部14为与该第二连接部43分别为可彼此互补嵌合的一环状的环凹槽141与一环状的凸垣431，并且该凹室20与该子室30皆位于该环凹槽141在该第一身部11上所围绕的范围内。当该第一本体10与该第二本体40相贴合时，该凸垣431及该环凹槽141相接合，借此使该凹室20与该子室30所构成的空间不与外界连通。此外，该环凹槽141与该子室30相隔开来而不相连通地作为一余液储存空间。而在其他实施态样，该余液储存空间有别于该环凹槽141，而另设于该第一身部11上，且不连通于该凹室20及该子室30。
本发明的该容器更包括更包括：一第三连接部15，设于该第一本体10相向于该第二本体40的一侧，并且位于该环凹槽141在该第一本体10上所围绕的范围之外的位置上；一第四连接部44，与该第三连接部15彼此间可分离或结合地设于该第二本体40的一侧，并且位于该凸垣431在该第二本体40上所围绕的范围之外的位置上；其中，当该第一本体10与该第二本体40相贴合时，该第三连接部15与该第四连接部44相互结合，可以使令该第二本体40与该第一本体10彼此相接。在本实施例中，该第三连接部15为与该第四连接部44分别为可彼此互补嵌合的一凹部151与一凸部441。其中，该凹部151可为但不限于一字形、T字形或十字形的突起结构，而该凸部441可为但不限于一字形、T字形或十字形的凹陷结构。
通过上述构件的组成，本发明的用以调节液体60分布的方法的具体实施步骤：
使液体60充盈于该凹室20与该子室30内，如图3所示。
施加一外力使该第二本体40的一端受自该第一本体10的第一端面12上分离，并朝着该第二本体40的另一端的方向逐步地远离该第一本体10。
如图4所示，在步骤B的施行期间，通过该液体60与该第二本体40间的接触，以该液体60与该第二本体40间的黏滞力F1，以及该液体60自身的表面张力F2，来克服该液体60的重力W，使位于该凹室20中的液体60随该第二本体40的远离而被带离，并使位于该子室30的液体60仍留存于该子室30内。其中， 该液体60的表面张力F2介于56.5dyne/cm～88.9dyne/cm之间，该液体60的黏度介于1.10cP～5.62cP之间，且该子室30的高度与该凹室20的高度比例介于12：1～3：5之间。在其他的实施方式中，随该第二本体40而被带离的液体60还可以利用于该第一本体10上所设的一流出口来排除。
该第二本体40完全自该第一本体10上卸除，使该子室30内残留有部分的液体60，而该环凹槽141则可以储存自该凹室20所带离的液体60，如图5A及图5B所示。
本发明可应用于培养细胞80的取用，其操作步骤如下，首先，该容器的该子室30中容纳有已培养成熟的细胞80，如图3的局部放大部分所示；接着，利用前揭所述的该方法的实施步骤，使原本通过该凹室20所充盈的液体60而相互连通的该些子室30，改变为各自独立且不相连通的状态(如图5A及图5B所示)，同时能达到不干扰容置于该子室30中的细胞80，以维持其所容纳的细胞80培养数量及状态。
以下将通过若干实例并搭配附图更进一步证实本发明所揭装置及方法的功效如后。
试验例1.液体60为0.1％的甲基纤维素溶液
首先，以0.1％的甲基纤维素配出4x10^4beads/ml(FluoSpheres TM F21010,15um,Invitrogen)，取1ul置入该些子室30中，再盖上该第二本体40后，静置10mins待其沉降。意即在试验例1中以被荧光标记的微珠取代细胞80进行实验。
接着，将0.1％的甲基纤维素溶液灌入该容器的容置空间中，其中，0.1％的甲基纤维素溶液的黏度为1.78cP。而后，在该第二本体40自该第一本体10上剥离的前、后分别观察该容器中四个预定区域内的微珠数量变化，并重复上述步骤以分别进行3组试验，其试验结果如表1、图6A、图6B、图7A、图7B、图8A及图8B所示，发现微珠数量并无明显的改变，且表1所揭的微珠数量些微的变动可能是被遮蔽所造成，故能在该第二本体40移除后仍有效地保持该子室30中的微珠数量及状态。
试验例2.液体60为0.3％的甲基纤维素溶液
与前述试验例1进行相同的试验步骤，而其差异在于灌入该容器的容置空间中的是0.3％的甲基纤维素溶液，其黏度为5.62cP，且其3组试验结果如表1、图9A、图9B、图10A、图10B、图11A及图11B所示，同样能有效地保持该子室30中的微珠培养数量及状态。
表1.第二本体剥离对容器内微珠数量的影响
Figure PCTCN2020088960-appb-000001
试验例3.液体为胰蛋白酶溶液
首先，CHO-K1 cell以CMFDA(CellTracker TM C7025,Invitrogen)进行染色(1:20)，配成4x10^4cell/ml，取1ul置入该些子室30中，再盖上该第二本体40，送入摄氏37度，5％CO2培养箱24hrs。接着，将胰蛋白酶溶液(Trypsin)灌入该容器的容置空间中，其中，胰蛋白酶溶液的黏度为1.1cP。而后，在该第二本体40自该第一本体10上剥离的前、后分别观察该容器内四个预定区域内的细胞80数量变化，并重复上述步骤分别进行3组试验，其试验结果如表2、图12A、图12B、图13A、图13B、图14A及图14B所示，发现细胞80数量并无明显的改变，且表2所揭的细胞80数量些微的变动可能是被遮蔽所造成，故能在该第二本体40移除后仍有效地保持该子室30中的细胞80数量及状态。此外，如图12C、图13C、及图14C所示，其为在该第二本体40剥离后，再将该容器内右上方观察区域内的细胞80吸除，作为阳性对照组。
试验例4.液体60为0.3％的F-12K培养基溶液
与前述试验例3进行相同的试验步骤，而其差异在于灌入该容器的容置空间中的是F-12K培养基溶液(F12k Growth Medium)，其黏度为1.1cP，且其3组试验结果如表2、图15A、图15B、图15C、图16A、图16B、图16C、图17A、图17B及图17C所示，同样能有效地保持该子室30中的细胞数量及状态。
表2.第二本体剥离对容器内细胞数量的影响
Figure PCTCN2020088960-appb-000002
Figure PCTCN2020088960-appb-000003
再者，如图18至图20所示，为本发明的第二实施例的示意图，而第二实施例与第一实施例的主要差异在于该子室30A凹设于该第一本体10A的第一端面12A，且该凹室20A对应该子室30A的位置而凹设于该第二本体40A的一侧，意即该凹室20A凹设于该第二本体40A的第二端面42A，且其壁面围绕形成该容纳空间50A。
接着，该第一本体10的第一端面12A上更凹设有一余液储存室16，其不连通于该子室30A，而分别与该环凹槽141A及该凹部151A连通，并且位于该环凹槽141A在该第一本体10A上所围绕的范围之外的位置上。
此外，该第二本体40A更包括有一凹陷部45，邻近该第二本体40A的端缘凹设于该第二本体40A的第二端面42A。当该第一本体10A与该第二本体40A相贴合时，使该凹部151A嵌合于该凸部441A中，以进一步固定该第一本体10A与该第二本体40A之间的连接关系，同时，该凹陷部45并不与该第一本体10A的第一端面12A相接触，而使该第一身部11A与该第二身部41A的相向端面之间存在有一余隙451，并与外部连通，借此便于使用者握持该第二本体40A，以利后续作动的进行。
另外，由于本例的具体实施步骤(如图21至图23所示)与第一实施例略同，故不再赘述。
如图24至图26所示，为本发明的第三实施例的示意图，而第三实施例与第二实施例的主要差异在于该容器更包括一第三本体70，当该第三本体70与该第一本体10B相连接时，将该第二本体40B夹设于该第一本体10B与该第三本体70之间。
其中，该容器还包括有可彼此分离或结合的一第五连接部17与一第六连接部71，分设于该第三本体70与该第一本体10B上，当该第三本体70与该第一本体10B相贴合时，使该第五连接部17与该第六连接部71相互结合，令该第三本体70结合于该第一本体10B上。在本实施例中，该第五连接部17为与该第六连接部71分别为可彼此互补嵌合的一凸部与一凹部，且该凸部与该凹部之间还能以点对点超音波熔接的方式连接，而使该凸部的至少一部分结合于该凹部的凹面上，可以将该第一本体10B与该第三本体70间暂时性地相互固定。
此外，将该第三本体70自该第一本体10B上卸除相当简便的，举例来说，可利用工具(例如：镊子)伸入该凸部与该凹部之间的缝隙中，再以该凹部的凹面为支点，并使该凸部的端面为着力点，而略微施力扳撬，即可顺势破坏该凸部与该凹部间的熔接关系，就能轻易的将前述接合结构撬开，达到快速简易拆卸的效用。

Claims (10)

  1. 一种用以调节液体分布的容器，其特征在于，包括：
    一第一本体；
    一子室，凹设于该第一本体的一侧；
    一具挠性的第二本体，可拆卸地覆于该第一本体上，可以封闭该子室；
    一容纳空间，被形成于该第一本体与该第二本体之间，并使该容纳空间与该子室彼此间能相互连通地容纳外部的液体；
    其中，当该第二本体的一端受外力驱使而自该第一本体的第一端面上掀开，并朝着该第二本体的另一端的方向逐步地远离该第一本体时，通过该液体表面与该第二本体间接触所产生的黏滞力，与该液体自身的表面张力，使位于该容纳空间中的液体随该第二本体的远离而被带离，并使位于该子室中的液体仍留存于该子室内。
  2. 如权利要求1所述用以调节液体分布的容器，其特征在于，更包括一凹室，对应该子室的位置而凹设于该第二本体的一侧，并由该凹室形成该容纳空间。
  3. 如权利要求1所述用以调节液体分布的容器，其特征在于，更包括一凹室，凹设于该第一本体的一侧，并且该子室凹设于该凹室底面，并连通该凹室所形成的该容纳空间。
  4. 如权利要求1所述用以调节液体分布的容器，其特征在于，更包括有可彼此分离或结合的一第一连接部与一第二连接部，分设于该第一本体与该第二本体之间彼此相向的端面上，当该第一本体与该第二本体相贴合时，该第一连接部与该第二连接部相互结合，可以使该第二本体与该第一本体彼此相接。
  5. 如权利要求4所述用以调节液体分布的容器，其特征在于，该第一连接部为与该第二连接部分别为可彼此互补嵌合的一环状的环凹槽与一环状的凸垣。
  6. 如权利要求5所述用以调节液体分布的容器，其特征在于，该第一本体更包括有一余液储存室，与该子室相间隔地凹设于该第一本体的一侧。
  7. 如权利要求5所述用以调节液体分布的容器，其特征在于，更包括：
    一第三连接部，设于该第一本体相向于该第二本体的一侧，并且位于该环凹槽在该第一本体上所围绕的范围之外的位置上；
    一第四连接部，与该第三连接部彼此间可分离或结合地设于该第二本体的一侧，并且位于该凸垣于该第二本体上所围绕的范围之外的位置上；
    其中，当该第一本体与该第二本体相贴合时，该第三连接部与该第四连接部相互结合，可以使令该第二本体与该第一本体彼此相接。
  8. 如权利要求1所述用以调节液体分布的容器，其特征在于，更包括一第三本体，当该第三本体与该第一本体相连接时，将该第二本体夹设于该第一本体与该第三本体之间。
  9. 如权利要求8所述用以调节液体分布的容器，其特征在于，更包括有可彼此分离或结合的一第五连接部与一第六连接部，分设于该第三本体与该第一本体上，当该第三本体与该第一本体相贴合时，使该第五连接部与该第六连接部相互结合，令该第三本体结合于该第一本体上。
  10. 一种用以调节液体分布的方法，其特征在于，使用权利要求1至9中任一项所述的用以调节液体分布的容器，使液体充盈于该容纳空间与该子室内，并施加一外力使该第二本体的一端自该第一本体上分离，并朝着该第二本体的另一端的方向逐步地远离该第一本体，通过该液体表面与该第二本体间接触所产生的黏滞力，以及该液体自身的表面张力，使位于该容纳空间中的液体随该第二本体的远离而被带离，并使位于该子室中的液体仍留存于该子室内。
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CN102858947A (zh) * 2010-04-15 2013-01-02 弗朗霍夫应用科学研究促进协会 细胞培养体系
CN105039139A (zh) * 2015-09-16 2015-11-11 烟台森森环保科技有限公司 一种微生物培养膜及微生物培养回收系统
WO2017091043A1 (ko) * 2015-11-26 2017-06-01 부산대학교 산학협력단 관류 세포 배양 마이크로 유체 시스템
CN108485964A (zh) * 2018-03-14 2018-09-04 苏杰 一种医用人体血红细胞培养仪
CN109337814A (zh) * 2018-11-07 2019-02-15 深圳大学 细胞孔板

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Publication number Priority date Publication date Assignee Title
CN102858947A (zh) * 2010-04-15 2013-01-02 弗朗霍夫应用科学研究促进协会 细胞培养体系
CN105039139A (zh) * 2015-09-16 2015-11-11 烟台森森环保科技有限公司 一种微生物培养膜及微生物培养回收系统
WO2017091043A1 (ko) * 2015-11-26 2017-06-01 부산대학교 산학협력단 관류 세포 배양 마이크로 유체 시스템
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