WO2021125406A1 - 정자 추출 마이크로 유체칩 및 그의 정자 추출 방법 - Google Patents
정자 추출 마이크로 유체칩 및 그의 정자 추출 방법 Download PDFInfo
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- WO2021125406A1 WO2021125406A1 PCT/KR2019/018197 KR2019018197W WO2021125406A1 WO 2021125406 A1 WO2021125406 A1 WO 2021125406A1 KR 2019018197 W KR2019018197 W KR 2019018197W WO 2021125406 A1 WO2021125406 A1 WO 2021125406A1
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- sperm
- fluid
- micropillars
- injection unit
- microfluidic chip
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502761—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502746—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the means for controlling flow resistance, e.g. flow controllers, baffles
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
- B01L2200/0652—Sorting or classification of particles or molecules
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0848—Specific forms of parts of containers
- B01L2300/0851—Bottom walls
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
- B01L2300/088—Channel loops
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
- B01L2400/0406—Moving fluids with specific forces or mechanical means specific forces capillary forces
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/08—Regulating or influencing the flow resistance
- B01L2400/084—Passive control of flow resistance
- B01L2400/086—Passive control of flow resistance using baffles or other fixed flow obstructions
Definitions
- the present invention relates to a sperm extraction microfluidic chip and a method for extracting sperm thereof, and more particularly, to a unidirectional flow of fluid using capillary effect and evaporation effect, and healthy sperm by using rheotaxis of sperm.
- the present invention relates to a sperm extraction microfluidic chip capable of effectively extracting spermatozoa, and a method for extracting sperm thereof.
- the serious low fertility problem is emerging as a global problem not only in Korea, but also infertility or infertility due to the decrease in fertility and genital diseases due to the aging of women and the increase in stress and environmental pollution are rapidly increasing.
- assisted reproductive techniques such as ovulation induction, intrauterine artificial insemination (IUI), in vitro fertilization, embryo transfer, and intracytoplasmic sperm injection have been developed and applied to actual clinical practice to overcome such infertility and infertility.
- a microfluidic chip was developed to separate healthy sperm, and even in this case, an external power source such as a fluid pump, syringe pump, etc. is required because the fluid flow is formed to separate the sperm, and thus the problem of complicated structure may occur.
- the embodiment of the present invention can form a unidirectional flow of fluid due to the capillary effect and evaporation effect generated in the micropillars without a separate external power source, and also allows sperm to move in the direction of the injection unit using mainstream properties, making it healthy
- a sperm extraction microfluidic chip capable of effectively extracting sperm is provided.
- the sperm extraction microfluidic chip includes a chip body having an injection unit formed on one side for injecting a fluid, disposed on the chip body to be symmetrical with a position of the injection unit, and the fluid injected into the injection unit includes a plurality of micropillars that generate a capillary effect and an evaporation effect so as to maintain a flow in one direction, and semen containing sperm is dropped onto the plurality of micropillars, and Some of the included sperm may be extracted from the injection unit by moving against the flow of the fluid by rheotaxis.
- a plurality of barrier rib channels may be provided between the plurality of micropillars and the injection unit to improve the flow of the sperms dropped onto the plurality of micropillars.
- the plurality of micropillars according to an embodiment of the present invention may be regularly arranged in horizontal and vertical directions to form a grid pattern.
- each of the plurality of micropillars according to an embodiment of the present invention may be formed as a square column, a rectangular column, or a rhombus-shaped column.
- each of the plurality of micropillars according to an embodiment of the present invention may be formed in a cylindrical or elliptical column.
- the plurality of barrier rib channels according to an embodiment of the present invention may be provided in parallel to each other so that a path is formed in the flow direction of the sperm.
- the plurality of barrier rib channels according to an embodiment of the present invention may be provided in a radially converging type or a pillar type in which a path is formed in the flow direction of the sperm and the width of the path becomes narrower toward the injection unit.
- the chip body is detachable from the chip body according to an embodiment of the present invention, and further comprises a cover part for covering the region of the injection part and the plurality of barrier rib channels, and the cover part injects the fluid into the injection part or the injection part A cover hole for extracting the collected sperm may be formed.
- the method for extracting sperm of a sperm extraction microfluidic chip includes: a fluid flow forming step of injecting a fluid into the injection unit to form a flow of the fluid in the one direction; A sperm transfer step of dropping semen containing sperm into a pillar so that some of the sperm included in the dropped semen move against the flow of the fluid by rheotaxis, and the sperm collected in the injection unit It may include a sperm extraction step of extracting them.
- a plurality of barrier rib channels are provided between the plurality of micropillars and the injection unit, and during the sperm movement step, the plurality of barrier rib channels cause the plurality of micropillars to pass through.
- the released sperm can improve the mainstreaming.
- the present invention it is possible to form a unidirectional flow of fluid due to the capillary effect and the evaporation effect generated in the micropillars without a separate external power source, and also the sperm can move in the direction of the injection part using the mainstream. It can effectively extract healthy sperm.
- FIG. 1 is a perspective view schematically illustrating a sperm extraction microfluidic chip according to an embodiment of the present invention.
- FIG. 2 is a perspective view in which a cover part is separated from the microfluidic chip shown in FIG. 1 .
- FIG. 3 is a plan view of the microfluidic chip shown in FIG. 2 excluding the cover.
- FIG. 4 is a flowchart illustrating a method for extracting sperm of the sperm extraction microfluidic chip shown in FIG. 1 .
- FIG. 5 is a plan view schematically illustrating a sperm extraction microfluidic chip according to another embodiment of the present invention.
- FIG. 1 is a perspective view schematically illustrating a sperm extraction microfluidic chip according to an embodiment of the present invention
- FIG. 2 is a perspective view of the microfluidic chip shown in FIG. 1 with a cover removed
- FIG. 3 is shown in FIG. It is a plan view excluding the cover part of the microfluidic chip.
- the sperm extraction microfluidic chip 100 includes a chip body 110 forming a basic frame, and a chip body 110 provided on the chip body 110 to inject fluid.
- the injection unit 120 the plurality of micro pillars 130 provided on the chip body 110 to form a flow of the fluid in one direction (arrow A direction), and the plurality of micro pillars 130 .
- It may include a plurality of partition wall channels 140 to improve rheotaxis so that sperm included in the dropped semen can go toward the injection unit 120 , and a cover unit 150 to cover the chip body 110 . have.
- the chip body 110 of this embodiment has a flat rectangular plate shape and an injection unit ( 120) is formed.
- the injection unit 120 may be provided with a fluid flowing in one direction within the chip body 110 , and as will be described later, sperm to be extracted are gathered in the injection unit 120 to extract sperm from the injection unit 120 . .
- separation prevention walls 121 are formed on the right and both sides of the injection unit 120 , so that the fluid injected into the injection unit 120 or the sperm collected in the injection unit 120 . It is possible to prevent the chip body 110 from being separated to the outside. Since the separation preventing wall 121 has a shape that gathers toward the right, when the fluid is injected into the injection unit 120, the injected fluid can be evenly spread to the partition wall channel 140 to be described later, and also the injection unit 120 direction. Sperms that move to the can also be well collected in the injection unit 120 .
- the fluid injected through the injection unit 120 may continuously flow in one direction (arrow B direction), that is, in the direction in which the plurality of micropillars 130 are provided in the injection unit 120 direction. That is, it is possible to maintain the flowability of the fluid.
- the plurality of micropillars 130 of the present embodiment may be arranged on the upper left side of the chip body 110 so as to be symmetrical to the position of the injection unit 120 as shown in FIGS. 1 to 3 .
- the plurality of micropillars 130 have a capillary effect and an evaporation effect so that the fluid injected into the injection unit 120 can maintain a flow in one direction (arrow A direction), that is, in the direction of the micropillars 130 . It has a structure that can be created.
- the plurality of micropillars 130 may be regularly arranged in horizontal and vertical directions.
- the plurality of micropillars 130 may be densely arranged to have a lattice pattern, and the capillary effect may be maximized due to this arrangement structure, so that the fluid may be drawn in one direction, and thus the flow of the fluid may be reduced. make it possible to keep
- the arrangement structure of the plurality of micropillars 130 has a grid pattern, but the present invention is not limited thereto. If the pattern can maximize the capillary effect and the evaporation effect, an arrangement structure other than the grid pattern may be applied. It is natural to have
- the plurality of micropillars 130 may have a square prism shape as shown in FIG. 3 , and thus each micropillar 130 is disposed close to the adjacent micropillars 130 to maximize the capillary effect. It can attract the fluid through it and make the fluid flow.
- the fluid injected into the injection unit 120 may continue to maintain flowability until the end of the micropillars 130 .
- the evaporation effect may be maximized to further induce evaporation of the fluid.
- the fluid can flow smoothly and continuously in one direction in the chip body 110 .
- the micropillar 130 may be provided in the shape of a square column as described above, but is not limited thereto.
- it may be provided in another shape such as a rectangular column, a rhombus-shaped column, a cylinder, or an elliptical column. is natural
- the flow of the fluid is formed as described above, and the flow of sperm is formed in a direction opposite to the flow of the fluid (arrow B direction) so that the sperm moves in the direction of the injection unit 120 , which will be described.
- the semen discharged from the male is dropped into the plurality of micropillars 130, and some of the sperm included in the dropped semen move in the direction of the injection unit 120 against the flow of the fluid due to mainstreaming. It may be extracted from the injection unit 120 .
- the sperm of the semen dropped onto the plurality of micropillars 130 are mixed with the fluid, and as described above, the fluid flows in the direction of the arrow A due to the capillary effect and evaporation effect generated by the micropillars 130 . and sperm move in the opposite direction (arrow B direction) against the fluid by mainstreaming. To put it simply, healthy sperm will try to move by swimming to the injection unit 120 by the mainstream.
- rheotaxis refers to the property of sperm to move in the opposite direction of the fluid flow when a fluid flows at a specific speed. Unhealthy sperm or white blood cells in semen can be swept away due to the flow of fluid, but healthy sperm can swim against the flow of fluid. This is to extract only healthy sperm.
- this embodiment has a structure for extracting sperm by the mainstream of the sperm, and without requiring an additional external power source for the flow of the fluid as in the prior art, the fluid flow can be maintained due to the capillary effect and the evaporation effect. convenient to have
- the microfluidic chip 100 of this embodiment may further include a plurality of barrier rib channels 140 so that healthy sperm can move well from the micropillars 130 to the injection unit 120 .
- the plurality of barrier rib channels 140 may be provided in parallel to each other to form a path in the flow direction of the sperm.
- healthy sperm has straightness, whereas unhealthy sperm rotates in place or draws a curve.
- the partition wall channel 140 of this embodiment allows healthy sperm to move straight in the direction of the injection unit 120 . ) to extract healthy sperm from In other words, the efficiency of extracting healthy sperm may be higher due to the plurality of partition wall channels 140 .
- the microfluidic chip 100 of this embodiment is detachable from the chip body 110 , and the region of the injection unit 120 and the plurality of partition wall channels 140 . It may further include a covering part 150 .
- a cover hole 151 for injecting a fluid into the injection unit 120 or extracting sperm collected in the injection unit 120 may be formed in the cover unit 150 . Due to the cover part 150 , it is possible to prevent the fluid or sperm moving in the chip body 110 from being affected by external foreign substances.
- FIG. 4 is a flowchart illustrating a method for extracting sperm of the sperm extraction microfluidic chip shown in FIG. 1 .
- the sperm extraction method of the sperm extraction microfluidic chip 100 includes a fluid flow forming step (S100), a sperm moving step (S200), and a sperm extraction step (S300). ) may be included.
- the fluid may be injected into the injection unit 120 to form a fluid flow in one direction.
- a capillary effect and an evaporation effect may occur when the fluid flows, and thus, the one-way flow of the fluid may be maintained.
- semen containing sperm is dropped into the plurality of micropillars 130, and some of the sperm included in the dropped semen are affected by the flow of the fluid due to rheotaxis. You can move backwards and backwards.
- healthy sperm that has been moved to the injection unit 120 due to mainstreaming may be extracted using an extractor (not shown).
- a one-way flow of fluid can be formed due to the capillary effect and evaporation effect generated in the micropillars 130 without a separate external power source, and also the sperm can use the mainstream. Since it can move in the direction of the injection unit 120, it is possible to effectively separate and extract healthy sperm.
- FIG. 5 is a plan view schematically illustrating a sperm extraction microfluidic chip according to another embodiment of the present invention.
- the microfluidic chip 200 according to another embodiment of the present invention is different from the fluid chip 100 (refer to FIG. 1 ) of the above-described embodiment in the structure of the barrier rib channel 240 .
- the barrier rib channel 240 of the present embodiment may be provided in a radially converging type in which a path is formed in the flow direction of the sperm and the width of the path becomes narrower toward the injection unit 220 direction. Therefore, since the sperm moving from the micropillars 230 to the injection unit 220 are moved to the injection unit 220 through a narrowing path, the mainstream of the sperm can be better used.
- the structure of the barrier rib channels 140 and 240 is not limited to the structure of the above-described embodiment or another embodiment, and it is natural that, for example, a pillar-type barrier rib channel may be provided.
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Abstract
Description
Claims (10)
- 일측에는 유체가 주입되는 주입부가 형성되는 칩본체; 및상기 주입부의 위치와 대칭되도록 상기 칩본체에 배치되며, 상기 주입부에 주입된 상기 유체가 일 방향으로의 흐름을 유지할 수 있도록 모세관 효과 및 증발 효과를 발생시키는 복수 개의 마이크로필라(Micro pillars);을 포함하며,정자가 포함된 정액이 상기 복수 개의 마이크로 필라에 투하되며, 투하된 상기 정액에 포함된 정자들의 일부 정자들이 주류성(rheotaxis)에 의해서 상기 유체의 흐름에 거슬러 이동하여 상기 주입부에서 추출되는 것을 특징으로 하는 정자 추출 마이크로 유체칩.
- 제1항에 있어서,상기 칩본체에서 상기 복수 개의 마이크로필라와 상기 주입부의 사이에는 상기 복수 개의 마이크로 필라에 투하된 상기 정자들이 주류성을 향상시키기 위한 복수 개의 격벽 채널이 구비되는 것을 특징으로 하는 정자 추출 마이크로 유체칩.
- 제1항에 있어서,상기 복수 개의 마이크로필라는 가로 및 세로 방향으로 규칙적으로 배치되어 격자 패턴을 형성하는 것을 특징으로 하는 정자 추출 마이크로 유체칩.
- 제3항에 있어서,상기 복수 개의 마이크로필라 각각은 정사각기둥, 직사각기둥 또는 마름모 형태의 기둥으로 형성되는 것을 특징으로 하는 정자 추출 마이크로 유체칩.
- 제3항에 있어서,상기 복수 개의 마이크로필라 각각은 원기둥 또는 타원기둥으로 형성되는 것을 특징으로 하는 정자 추출 마이크로 유체칩.
- 제2항에 있어서,상기 복수 개의 격벽 채널은 상기 정자의 흐름 방향으로 경로가 형성되도록 상호 평행하게 마련되는 것을 특징으로 하는 정자 추출 마이크로 유체칩.
- 제2항에 있어서,상기 복수 개의 격벽 채널은 상기 정자의 흐름 방향으로 경로가 형성되되 상기 주입부 방향으로 갈수록 경로의 폭이 좁아지는 방사 수렴형으로 마련되거나 필라 타입으로 마련되는 것을 특징으로 하는 정자 추출 마이크로 유체칩.
- 제2항에 있어서,상기 칩본체에 착탈 가능하며, 상기 주입부의 영역 및 상기 복수 개의 격벽 채널을 덮는 덮개부를 더 포함하며,상기 덮개부에는 상기 주입부에 상기 유체를 주입하거나 상기 주입부에 모아진 상기 정자들을 추출하기 위한 덮개홀이 형성되는 것을 특징으로 하는 정자 추출 마이크로 유체칩.
- 일측에는 유체가 주입되는 주입부가 형성되는 칩본체와, 상기 주입부의 위치와 대칭되도록 상기 칩본체에 배치되며 상기 주입부에 주입된 상기 유체가 일 방향으로의 흐름을 유지할 수 있도록 모세관 효과 및 증발 효과를 발생시키는 복수 개의 마이크로필라를 포함하는 정자 추출 마이크로 유체칩의 정자 추출 방법에 있어서,상기 주입부에 유체를 주입하여 상기 일 방향으로의 상기 유체의 흐름을 형성하는 유체 흐름 형성 단계;상기 복수 개의 마이크로필라에 정자가 포함된 정액을 투하하여, 투하된 상기 정액에 포함된 정자들의 일부 정자들이 주류성(rheotaxis)에 의해서 상기 유체의 흐름에 거슬러 이동하도록 하는 정자 이동 단계; 및상기 주입부에 모아진 상기 정자들을 추출하는 정자 추출 단계;를 포함하는 것을 특징으로 하는 정자 추출 마이크로 유체칩의 정자 추출 방법.
- 제9항에 있어서,상기 칩본체에서 상기 복수 개의 마이크로필라와 상기 주입부의 사이에는 복수 개의 격벽 채널이 구비되며,상기 정자 이동 단계 시, 상기 복수 개의 격벽 채널로 인해 상기 복수 개의 마이크로필라에 투하된 상기 정자들이 주류성이 향상되는 것을 특징으로 하는 정자 추출 마이크로 유체칩의 정자 추출 방법.
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CN113528310A (zh) * | 2021-06-16 | 2021-10-22 | 复旦大学 | 一种模拟宫颈微环境的仿生微流控芯片及其制备方法 |
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AR107746A1 (es) * | 2017-02-24 | 2018-05-30 | Herberto Ernesto Hector Repetto | Dispositivo y método de separación de células móviles |
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