WO2020192168A1 - Microfluidic chip and in vitro testing device containing the microfluidic chip - Google Patents

Microfluidic chip and in vitro testing device containing the microfluidic chip Download PDF

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Publication number
WO2020192168A1
WO2020192168A1 PCT/CN2019/122795 CN2019122795W WO2020192168A1 WO 2020192168 A1 WO2020192168 A1 WO 2020192168A1 CN 2019122795 W CN2019122795 W CN 2019122795W WO 2020192168 A1 WO2020192168 A1 WO 2020192168A1
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cavity
microfluidic chip
flow channel
sample
quantitative
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PCT/CN2019/122795
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French (fr)
Chinese (zh)
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蒙玄
刘洋
万惠芳
李文美
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广州万孚生物技术股份有限公司
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    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027Containers 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
    • B01L3/502707Containers 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 manufacture of the container or its components
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/12Specific details about manufacturing devices
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/08Geometry, shape and general structure
    • B01L2300/0803Disc shape
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N2035/00465Separating and mixing arrangements
    • G01N2035/00495Centrifuges

Definitions

  • the invention relates to the technical field of in vitro diagnostics, in particular to a microfluidic chip and an in vitro detection device containing the microfluidic chip.
  • In Vitro Diagnosis refers to the technology of taking samples (blood, body fluids, tissues, etc.) from the human body for detection and analysis to diagnose diseases.
  • the detection process requires corresponding instruments and reagents, and these instruments and reagents It constitutes an in vitro diagnostic system.
  • In-vitro diagnostic systems are roughly divided into two types: one is represented by the testing center laboratory, which has the characteristics of system modularization and automation, and conducts pipeline inspections on samples, which also has high throughput, high efficiency, and high sensitivity.
  • the entire system also has disadvantages such as high cost, large volume, and professional operation.
  • POCT point-of-care testing
  • microfluidic chip with a relatively simple structure and capable of separating and quantifying samples, and an in vitro detection device containing the microfluidic chip.
  • a microfluidic chip is provided with a separation and quantification unit.
  • the separation and quantification unit includes a sample addition cavity, a first microchannel, a precipitation cavity, a capillary channel, and a second microchannel And a quantitative cavity;
  • the sample loading cavity is provided with a sample loading hole, the sample loading cavity and the precipitation cavity are communicated through the first micro flow channel, and the first micro flow channel passes through the capillary
  • the flow channel is in communication with the second micro flow channel, and the second micro flow channel is in communication with the quantitative cavity;
  • the microfluidic chip has a center of rotation, the precipitation chamber is farther from the center of rotation than the sample loading chamber, and the capillary channel is closer to the center of rotation after being connected to the first microchannel. It extends in the direction of and bends and then extends in a direction away from the rotation center to connect with the second micro flow channel, and the quantitative cavity is farther from the rotation center than the capillary flow channel.
  • the separation and quantification unit further includes a waste liquid cavity, the waste liquid cavity is in communication with the second microchannel, and the waste liquid cavity is located on the second microchannel. Downstream of the quantitative cavity, the waste liquid cavity is farther from the rotation center than the capillary flow channel.
  • the sample adding cavity is further provided with a first vent hole.
  • a baffle is provided in the sample loading cavity between the sample loading hole and the first vent hole.
  • the sample adding hole and the first vent hole are both arranged on the sample adding cavity close to the rotation center.
  • the width of the capillary flow channel is 0.1 mm to 0.2 mm, and the depth is 0.1 mm to 0.2 mm; or
  • the width of the capillary flow channel is 0.2mm-0.5mm, and the depth is 0.2mm-0.5mm, and the flow channel wall of the capillary flow channel is surface treated with PEG4000.
  • the second microfluidic channel is provided with a second ventilation hole, the second ventilation hole is located downstream of the cavity structure connected to the second microfluidic channel, and the second ventilation hole Relative to the cavity structure connected to the second micro flow channel, it is closer to the rotation center.
  • the portion of the second micro-channel downstream of the cavity structure to which it is connected is bent and extended in a direction close to the rotation center, and the second vent is provided in the second micro-channel. The end of the runner.
  • the microfluidic chip is in the shape of a disc, a plurality of the separation and quantitative units are evenly distributed on the microfluidic chip, and the middle of the microfluidic chip has a rotating mounting part, so The center of the rotation mounting part is the rotation center.
  • the microfluidic chip includes a chip body and a transparent cover film covering the chip body, and the chip body and the transparent cover film cooperate to form each cavity of the separation and quantitative unit Structure and runner structure.
  • An in vitro detection device includes the microfluidic chip described in any of the above embodiments and a detection unit, and the detection unit is used to detect a sample in the quantitative cavity.
  • the detection unit is a freeze-dried reagent set in the quantitative cavity.
  • the quantitative cavity has a permeation hole, the permeation hole is covered with a water-soluble film, and the sampling port of the detection unit is connected to the permeation hole and is separated by the water-soluble film .
  • the detection unit is a dry chemical test paper.
  • the dry chemical test paper includes a support layer and a reaction indicator layer and a diffusion layer sequentially stacked on the support layer, and the reaction indicator layer contains a substance capable of reacting with the target substance in the sample to be tested. Reaction reagents and indicator reagents, and the diffusion layer faces the water-soluble membrane through the injection port.
  • the microfluidic chip is provided with a mounting groove around the penetration hole, and the detection unit is embedded in the mounting groove.
  • the microfluidic chip is equipped with a sample loading cavity, a sedimentation cavity and a quantitative cavity.
  • the sample to be tested can be added to the sample loading cavity through the sample hole, and the solid precipitate can be separated from the liquid by centrifugal separation.
  • the test solution containing the target substance the test solution in the sample loading chamber and the first microchannel can drive the liquid to flow through the capillary force of the capillary channel, and finally form a siphon effect and external centrifugal action to flow into the quantitative cavity to achieve the Measure the quantification of the solution.
  • the microfluidic chip has a relatively simple structure, is easy to manufacture and shape, and can be widely promoted and used.
  • the in-vitro detection device has a detection unit, which can directly detect the quantitative solution to be tested in the quantitative cavity, and has simple operation and high detection efficiency.
  • FIG. 1 is a schematic diagram of the front structure of a microfluidic chip according to an embodiment of the present invention
  • FIG. 2 is a schematic diagram of the backside structure of the microfluidic chip shown in FIG. 1;
  • Fig. 3 is a side view of the microfluidic chip shown in Fig. 1 with a detection unit;
  • Figures 4-1, 4-2, 4-3 and 4-4 are schematic diagrams of the process of plasma (or serum) separation and quantification of the whole blood sample by the microfluidic chip shown in Figure 1, Figure 4-2-1, 4 -3-1 and 4-4-1 are corresponding partial enlarged schematic diagrams;
  • FIGS. 5-1, 5-2, and 5-3 are schematic diagrams of the process of dissolving the water-soluble membrane into the detection unit by plasma (or serum) of an in vitro detection device according to an embodiment of the present invention
  • FIG. 6 is a schematic diagram of the structure of a dry chemical test paper according to an embodiment.
  • a component when referred to as being “disposed on” or “mounted on” another component, it can be directly on the other component or a centered component may also exist.
  • an element When an element is considered to be “connected” to another element, it can be directly connected to the other element or an intermediate element may be present at the same time.
  • an embodiment of the present invention provides a microfluidic chip 10 on which a separation and quantification unit 100 is provided.
  • the separation and quantification unit 100 includes a sample addition cavity 110, a first micro flow channel 120, a sedimentation cavity 130, a capillary flow channel 140, a second micro flow channel 150 and a quantification cavity 160.
  • the sample adding cavity 110 is used to hold the sample to be tested, and is provided with a sample adding hole 111.
  • the sedimentation chamber 130 is used to collect useless objects in the sample to be tested with a relatively high density of solids.
  • the sample loading cavity 110 and the sedimentation cavity 130 are in communication with each other through the first micro flow channel 120.
  • the first micro channel 120 communicates with the second micro channel 150 through the capillary channel 140.
  • the second micro flow channel 150 is in communication with the quantitative cavity 160.
  • the quantitative cavity 160 is used to quantify the sample solution to be tested.
  • the microfluidic chip 10 has a center of rotation 18.
  • the sedimentation cavity 130 is farther away from the rotation center 18 than the sample addition cavity 110.
  • the capillary flow channel 140 extends from the connection with the first micro flow channel 120 in the direction approaching the rotation center 18 (may be each gradually approaching the rotation center 18, such as but not limited to the radial direction), and then bends away from the rotation center.
  • the direction of 18 (may be a direction gradually away from the rotation center 18, for example, but not limited to a radial direction) extends to connect with the second micro channel 150.
  • the quantitative cavity 160 is farther from the center of rotation than the capillary flow channel 140.
  • the bottom of the end connecting the sample adding cavity 110 and the first micro flow channel 120 is inclined to facilitate the flow of the sample to be tested into the first micro flow channel 120.
  • the size of the first microfluidic channel 120 needs to be able to pass through the useless objects in the sample to be tested. For example, for a whole blood sample, when separating plasma or serum, the first microfluidic channel 120 needs to satisfy Of blood cells can pass through.
  • the first micro flow channel 120 has a branch portion 121 extending in a direction close to the rotation center 18, and the capillary flow channel 140 is connected to the end of the branch portion 121 of the first micro flow channel 120.
  • the separation and quantitative unit 100 further includes a waste liquid cavity 170.
  • the waste liquid chamber 170 is used to collect excess solution to be tested.
  • the waste liquid cavity 170 is in communication with the second micro flow channel 150.
  • the waste liquid cavity 170 is located downstream of the quantitative cavity 160 on the second micro flow channel 150 to receive the excess solution to be tested.
  • the waste liquid cavity 170 is farther away from the rotation center 18 than the capillary flow channel 140.
  • the sample adding cavity 110 is further provided with a first vent 112.
  • the first vent hole 112 is used for ventilation, which can facilitate the addition of the sample, and avoid the influence of the sample entry due to the increase of the air pressure inside the cavity when the sample is added.
  • a baffle 113 is provided between the sample adding hole 111 and the first vent hole 112 in the sample adding cavity 110.
  • the baffle 113 can prevent the sample from reaching one side of the first vent hole 112 after the sample is added, and prevent the sample from flowing out of the first vent hole 112.
  • the sample addition hole 111 and the first vent hole 112 are both arranged on the sample addition cavity 110 close to the rotation center 18, so that when the sample is rotated and centrifuged to make the sample flow to the side of the sample addition cavity 110 far from the rotation center, The sample is placed to flow out from the sample loading hole 111 and the first vent hole 112, so that the sample flows smoothly into the first micro flow channel 120 and the sedimentation cavity 130.
  • the capillary flow channel 140 has a V shape, and its curved part is close to the rotation center 18.
  • the capillary flow channel 140 has a width of 0.1 mm to 0.2 mm and a depth of 0.1 mm to 0.2 mm; or the capillary flow channel 140 has a width of 0.2 mm to 0.5 mm and a depth of 0.2 mm to 0.5 mm.
  • no surface treatment is required.
  • the flow channel wall of the capillary flow channel 140 is preferably surface-treated with PEG4000. Further preferably, the width of the capillary flow channel 140 is 0.2 mm, and the depth is also 0.2 mm. After the capillary channel 140 enters the sample solution, the sample solution can flow to the other end of the sample solution by capillary action, and finally form a siphon effect between the first micro channel 120 and the second micro channel 150.
  • the PEG4000 surface treatment can be, but is not limited to, adding a 1 wt% PEG4000 solution to the capillary flow channel 140 and forming it after natural drying.
  • the surface treatment of PEG4000 is beneficial to increase the capillary force of the capillary channel 140, and PEG4000 is an inert substance in the reaction system, and generally does not react with samples and detection reagents, and thus does not affect the detection results.
  • the second micro flow channel 150 is provided with a second vent hole 151.
  • the second vent hole 151 is located downstream of the cavity structure (for example, the quantitative cavity 160 and the waste liquid cavity 170) connected to the second micro flow channel 150, and the second vent hole 151 is connected to the second micro flow channel 150
  • the cavity structure is closer to the center of rotation 18.
  • the second vent hole 151 also plays a role of ventilating, facilitating the sample solution to be tested to smoothly flow into the second micro flow channel 150 and finally flow into the quantitative cavity 160 and the waste liquid cavity 170.
  • first vent 112 and the second vent 151 can be selected.
  • the part of the second micro-channel 150 downstream of the cavity structure connected to it is bent and extended in a direction close to the rotation center 18, and the second vent hole 151 is provided at the end of the second micro-channel 150, which can effectively The sample solution is prevented from flowing out from the second vent hole 151.
  • the microfluidic chip 10 includes a chip body 11 and a transparent cover film 12 covering the chip body 11.
  • the chip body 11 and the transparent cover film 12 cooperate to form the cavity structure and the flow channel structure of the separation and quantitative unit 100.
  • the grooves of each cavity structure and flow channel structure are pre-formed on the chip body 11.
  • each hole is opened on the back of the chip body, and is subsequently covered by a transparent cover film 12 and sealed in
  • the front surface of the chip body 11 can be formed to complete the packaging of the cavity structure and the flow channel structure, forming a complete cavity structure and the flow channel structure.
  • the transparent cover film 12 can be, but is not limited to, transparent tape or transparent pressure-sensitive adhesive, etc., which cooperates with the chip body 11 to form the entire microfluidic chip 10, which is simple to assemble and does not require complicated and expensive ultrasonic welding technology. , Can significantly reduce production costs. It can be understood that, in other specific examples, the microfluidic chip 10 may also be formed by welding with a relatively high-cost ultrasonic welding technology, or be integrally formed with a 3D printing technology.
  • multiple separation and quantitative units 100 there are multiple separation and quantitative units 100, and the multiple separation and quantitative units 100 are arranged around the same rotation center 18.
  • Multiple quantification units 100 can realize multi-sample single detection, and can also realize single-sample multiple detection, with good consistency and high integration, which significantly improves the throughput of single detection.
  • the microfluidic chip 10 is in the shape of a disc, a plurality of separate quantitative units 100 are evenly distributed on the microfluidic chip 10, and the microfluidic chip 10 has a rotating mounting part 180 in the middle.
  • the center of the rotating mounting portion 180 is the center of rotation 18.
  • the rotating mounting part 180 may be various types of clamping slots or clamping posts.
  • the microfluidic chip 10 is provided with a sample loading cavity 110, a precipitation cavity 130, and a quantitative cavity 160.
  • the sample to be tested can be added to the sample loading cavity 110 through the sample hole 111, and the solid can be precipitated by centrifugal separation.
  • the substance is separated from the liquid to obtain the test solution containing the target substance.
  • the test solution in the sample loading chamber 110 and the first microchannel 120 can drive the liquid to flow through the capillary force of the capillary channel 140, and finally form a siphon effect and the outside world.
  • the centrifugal action flows into the quantification cavity 160 to realize the quantification of the solution to be tested.
  • the microfluidic chip 10 has a relatively simple structure, is easy to manufacture and shape, and can be widely promoted and used.
  • the present invention also provides an in vitro detection device 2, which includes the aforementioned microfluidic chip 10 and a detection unit 20.
  • the detection unit 20 is used to detect the sample in the quantitative cavity 160 of the microfluidic chip 10.
  • the detection unit 20 is a freeze-dried reagent arranged in the quantitative cavity 160.
  • the freeze-dried reagent is placed in the quantitative cavity 160.
  • the freeze-dried reagent can be dissolved and reacted with it, and the result of the reaction can be detected.
  • the detection unit 20 is external. Specifically, the quantitative cavity 160 has a permeation hole 161, and the permeation hole 161 is covered with a water-soluble film 162. The injection port 21 of the detection unit 20 is connected to the permeation hole 161 and is separated by a water-soluble membrane 162.
  • the detection unit 20 is a dry chemical test paper.
  • the dry chemical test paper 20 includes a supporting layer 22 and a reaction indicating layer 23 and a diffusion layer 24 stacked on the supporting layer 22 in sequence.
  • the reaction indicator layer 23 contains a reaction reagent and an indicator reagent that can react with the target substance in the sample to be tested.
  • the reaction indicating layer 23 may be one layer or multiple layers.
  • the reaction indicating layer 23 includes two layers of an indicating layer 231 and a reagent layer 232, and the indicating layer 231 is close to the supporting layer 22.
  • the reagent layer 232 is close to the diffusion layer 24, and contains a reaction reagent capable of reacting with the target substance; in addition, the reagents contained in the indicator layer 232 and the reagent layer 232 can also be exchanged or appropriately mixed.
  • the diffusion layer 24 faces the water-soluble film 162 through the injection port 21.
  • microfluidic chip 10 is provided with a mounting groove around the penetration hole.
  • the detection unit 20 is embedded in the installation groove.
  • the in vitro detection device 2 has a detection unit 20, which can directly detect the quantified sample solution to be tested in the quantitative cavity 160, with simple operation and high detection efficiency.
  • a detection unit 20 which can directly detect the quantified sample solution to be tested in the quantitative cavity 160, with simple operation and high detection efficiency.
  • the specific test process of the in vitro test device 2 using dry chemical test paper can be referred to as follows:
  • the rotary mounting part 180 of the in vitro detection device 2 After the sample is added, install the rotary mounting part 180 of the in vitro detection device 2 in the supporting rotary centrifugal instrument, turn on the instrument to rotate, the whole blood is subjected to centrifugal force, and the red blood cells are deposited in the sedimentation chamber 130, and the serum or plasma is separated To the upper part of the precipitation chamber 130, the second micro flow channel 120, and the sample adding chamber 110;
  • the microfluidic chip 10 When the capillary channel 140 is filled with serum or plasma, the microfluidic chip 10 is controlled to rotate again, because the end 140c is farther from the center of rotation 18 than the liquid level in the sample cavity 110, as shown in Figures 4-3-1 and 4- As shown in 4-1, under the action of siphoning and centrifugal force, serum or plasma enters the quantitative cavity 160 through the second microchannel 150.
  • the sample solution to be tested is completed Quantitatively, the excess serum or plasma enters the back section of the second micro-channel 150 or the waste liquid cavity 170; preferably, the instrument will determine whether there is in the back section of the second micro-channel 150 or the waste liquid cavity 170.
  • the liquid judges whether the quantitative cavity 160 is full of liquid. When it is detected that there is liquid in the back section of the second microfluidic channel 150 or the waste liquid cavity 170, it can be determined that the quantitative cavity 160 is full of liquid. Whether the cavity 160 is full of liquid;
  • the liquid generally does not flow into the detection unit (dry chemical test paper) 20 due to its own weight.
  • the microfluidic chip 10 needs to be rotated under control. It can rotate at a low speed of 1800rpm ⁇ 2000rpm to make the quantitative chamber
  • the quantified sample solution in 160 enters the sample inlet 21 of the detection unit 20;
  • the sample solution is sequentially diffused through the diffusion layer 24 to the reaction indicator layer 23 for color reaction.
  • the depth of the color can reflect the concentration of the test substance.
  • the signal can be collected through the detection hole 25 of the detection unit, and finally converted into the concentration data of the test substance. .
  • the in vitro detection device 2 can solve the problems of low sensitivity and stability of dry chemical test paper whole blood, and has the advantages of high detection throughput and low cost.

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Abstract

A microfluidic chip (10) and an in vitro testing device (2) containing the microfluidic chip (10). The microfluidic chip (10) is provided with a sample adding cavity (110), a precipitation cavity (130) and a quantification cavity (160); a sample to be tested can be added to the sample adding cavity (110) through a sample adding hole (111), a solid precipitate can be separated from a liquid by means of centrifugal separation, so as to obtain a solution to be tested containing a target substance, and said solution in the sample adding cavity (110) and a first microfluidic channel (120) can drive the liquid to flow by means of the capillary force of a capillary flow channel (140), finally forming a siphoning action and an external centrifugal action to enable said solution to flow into the quantification cavity (160), so as to achieve the quantification of said solution. The microfluidic chip (10) has a relatively simple structure, is easy to manufacture and mold, and can be widely promoted and used. The in vitro testing device (2) has a testing unit (20) and can directly test a solution to be tested which has been quantified in a quantification cavity (160), being easy to operate and having high testing efficiency.

Description

微流控芯片及含有该微流控芯片的体外检测装置Microfluidic chip and in vitro detection device containing the microfluidic chip 技术领域Technical field
本发明涉及体外诊断技术领域,尤其是涉及一种微流控芯片及含有该微流控芯片的体外检测装置。The invention relates to the technical field of in vitro diagnostics, in particular to a microfluidic chip and an in vitro detection device containing the microfluidic chip.
背景技术Background technique
体外诊断(In Vitro Diagnosis,IVD)是指从人体中取出样本(血液、体液、组织等)进行检测分析从而对疾病进行诊断的技术,检测过程中需要相应的仪器和试剂,而这些仪器和试剂就组成了体外诊断系统。体外诊断系统大致分为两种:一种是以检测中心实验室为代表,它具有系统模块化、自动化的特点,对样本进行流水线式的检验,从而也具有高通量、高效率、高敏感度的优势,但是整套系统也存在费用昂贵、所占体积大、需要专业人员操作等缺陷,它主要应用于大型医院;另外一种是以即时检测(point-of-care testing,POCT)为代表,它的系统具有集成化、小型化的特点,可随时随地进行样本检验,从而也具有价格实惠、操作简单、结果报告及时的优势。微流控芯片的技术的优势就是功能集成度高、小型化、自动化,因此微流控技术在POCT领域中得到广泛的应用。然而,传统的用于POCT的微流控芯片普遍存在结构复杂的问题,因而价格相对昂贵,限制了其在POCT领域的进一步应用。In Vitro Diagnosis (IVD) refers to the technology of taking samples (blood, body fluids, tissues, etc.) from the human body for detection and analysis to diagnose diseases. The detection process requires corresponding instruments and reagents, and these instruments and reagents It constitutes an in vitro diagnostic system. In-vitro diagnostic systems are roughly divided into two types: one is represented by the testing center laboratory, which has the characteristics of system modularization and automation, and conducts pipeline inspections on samples, which also has high throughput, high efficiency, and high sensitivity. However, the entire system also has disadvantages such as high cost, large volume, and professional operation. It is mainly used in large hospitals; the other is represented by point-of-care testing (POCT) , Its system has the characteristics of integration and miniaturization, which can carry out sample inspection anytime and anywhere, which also has the advantages of affordable price, simple operation, and timely result reporting. The advantages of microfluidic chip technology are high functional integration, miniaturization, and automation. Therefore, microfluidic technology is widely used in the POCT field. However, the traditional microfluidic chip used for POCT generally has the problem of complex structure, and therefore the price is relatively expensive, which limits its further application in the POCT field.
发明内容Summary of the invention
基于此,有必要提供一种结构相对简单且可实现对样本分离和定量的微流 控芯片及含有该微流控芯片的体外检测装置。Based on this, it is necessary to provide a microfluidic chip with a relatively simple structure and capable of separating and quantifying samples, and an in vitro detection device containing the microfluidic chip.
一种微流控芯片,所述微流控芯片上设有分离定量单元,所述分离定量单元包括加样腔体、第一微流道、沉淀腔体、毛细流道、第二微流道和定量腔体;所述加样腔体设有加样孔,所述加样腔体与所述沉淀腔体通过所述第一微流道连通,所述第一微流道通过所述毛细流道与所述第二微流道连通,所述第二微流道与所述定量腔体连通;A microfluidic chip. The microfluidic chip is provided with a separation and quantification unit. The separation and quantification unit includes a sample addition cavity, a first microchannel, a precipitation cavity, a capillary channel, and a second microchannel And a quantitative cavity; the sample loading cavity is provided with a sample loading hole, the sample loading cavity and the precipitation cavity are communicated through the first micro flow channel, and the first micro flow channel passes through the capillary The flow channel is in communication with the second micro flow channel, and the second micro flow channel is in communication with the quantitative cavity;
所述微流控芯片具有旋转中心,所述沉淀腔体较所述加样腔体远离所述旋转中心,所述毛细流道自与所述第一微流道连接后向靠近所述旋转中心的方向延伸并弯曲后向远离所述旋转中心的方向延伸以与所述第二微流道连接,所述定量腔体较所述毛细流道远离所述旋转中心。The microfluidic chip has a center of rotation, the precipitation chamber is farther from the center of rotation than the sample loading chamber, and the capillary channel is closer to the center of rotation after being connected to the first microchannel. It extends in the direction of and bends and then extends in a direction away from the rotation center to connect with the second micro flow channel, and the quantitative cavity is farther from the rotation center than the capillary flow channel.
在其中一个实施例中,所述分离定量单元还包括废液腔体,所述废液腔体与所述第二微流道连通,所述废液腔体在所述第二微流道上位于所述定量腔体的下游,所述废液腔体较所述毛细流道远离所述旋转中心。In one of the embodiments, the separation and quantification unit further includes a waste liquid cavity, the waste liquid cavity is in communication with the second microchannel, and the waste liquid cavity is located on the second microchannel. Downstream of the quantitative cavity, the waste liquid cavity is farther from the rotation center than the capillary flow channel.
在其中一个实施例中,所述加样腔体还设有第一透气孔。In one of the embodiments, the sample adding cavity is further provided with a first vent hole.
在其中一个实施例中,所述加样腔体内在所述加样孔与所述第一透气孔之间设有阻流板。In one of the embodiments, a baffle is provided in the sample loading cavity between the sample loading hole and the first vent hole.
在其中一个实施例中,所述加样孔及所述第一透气孔在所述加样腔体上均靠近所述旋转中心设置。In one of the embodiments, the sample adding hole and the first vent hole are both arranged on the sample adding cavity close to the rotation center.
在其中一个实施例中,所述毛细流道的宽度为0.1mm~0.2mm,深度为0.1mm~0.2mm;或者In one of the embodiments, the width of the capillary flow channel is 0.1 mm to 0.2 mm, and the depth is 0.1 mm to 0.2 mm; or
所述毛细流道的宽度为0.2mm~0.5mm,深度为0.2mm~0.5mm,所述毛细流道的流道壁经PEG4000表面处理。The width of the capillary flow channel is 0.2mm-0.5mm, and the depth is 0.2mm-0.5mm, and the flow channel wall of the capillary flow channel is surface treated with PEG4000.
在其中一个实施例中,所述第二微流道上设有第二透气孔,所述第二透气孔位于所述第二微流道上连接的腔体结构的下游,且所述第二透气孔相对于所述第二微流道上连接的腔体结构更靠近于所述旋转中心。In one of the embodiments, the second microfluidic channel is provided with a second ventilation hole, the second ventilation hole is located downstream of the cavity structure connected to the second microfluidic channel, and the second ventilation hole Relative to the cavity structure connected to the second micro flow channel, it is closer to the rotation center.
在其中一个实施例中,所述第二微流道上位于其连接的腔体结构的下游的部分向靠近所述旋转中心的方向弯折延伸,所述第二透气孔设于所述第二微流道的末端。In one of the embodiments, the portion of the second micro-channel downstream of the cavity structure to which it is connected is bent and extended in a direction close to the rotation center, and the second vent is provided in the second micro-channel. The end of the runner.
在其中一个实施例中,所述分离定量单元有多个,多个所述分离定量单元围绕同一所述旋转中心设置。In one of the embodiments, there are multiple separation and quantitative units, and multiple separation and quantitative units are arranged around the same rotation center.
在其中一个实施例中,所述微流控芯片呈圆盘状,多个所述分离定量单元均匀分布在所述微流控芯片上,所述微流控芯片的中部具有旋转安装部,所述旋转安装部的中心即所述旋转中心。In one of the embodiments, the microfluidic chip is in the shape of a disc, a plurality of the separation and quantitative units are evenly distributed on the microfluidic chip, and the middle of the microfluidic chip has a rotating mounting part, so The center of the rotation mounting part is the rotation center.
在其中一个实施例中,所述微流控芯片包括芯片本体和覆盖在所述芯片本体上的透明盖膜,所述芯片本体与所述透明盖膜配合形成所述分离定量单元的各腔体结构和流道结构。In one of the embodiments, the microfluidic chip includes a chip body and a transparent cover film covering the chip body, and the chip body and the transparent cover film cooperate to form each cavity of the separation and quantitative unit Structure and runner structure.
一种体外检测装置,包括上述任一实施例所述的微流控芯片和检测单元,所述检测单元用于检测所述定量腔体内的样本。An in vitro detection device includes the microfluidic chip described in any of the above embodiments and a detection unit, and the detection unit is used to detect a sample in the quantitative cavity.
在其中一个实施例中,所述检测单元为设置在所述定量腔体内的冻干试剂。In one of the embodiments, the detection unit is a freeze-dried reagent set in the quantitative cavity.
在其中一个实施例中,所述定量腔体具有渗透孔,所述渗透孔上覆盖有水溶性膜,所述检测单元的进样口与所述渗透孔对接且由所述水溶性膜隔开。In one of the embodiments, the quantitative cavity has a permeation hole, the permeation hole is covered with a water-soluble film, and the sampling port of the detection unit is connected to the permeation hole and is separated by the water-soluble film .
在其中一个实施例中,所述检测单元为干化学试纸。In one of the embodiments, the detection unit is a dry chemical test paper.
在其中一个实施例中,所述干化学试纸包括支撑层和在所述支撑层上依次层叠设置的反应指示层和扩散层,所述反应指示层中含有能够与待测样本中目 标物质反应的反应试剂和指示试剂,所述扩散层通过所述进样口面向于所述水溶性膜。In one of the embodiments, the dry chemical test paper includes a support layer and a reaction indicator layer and a diffusion layer sequentially stacked on the support layer, and the reaction indicator layer contains a substance capable of reacting with the target substance in the sample to be tested. Reaction reagents and indicator reagents, and the diffusion layer faces the water-soluble membrane through the injection port.
在其中一个实施例中,所述微流控芯片围绕所述渗透孔设有安装槽,所述检测单元镶嵌在所述安装槽中。In one of the embodiments, the microfluidic chip is provided with a mounting groove around the penetration hole, and the detection unit is embedded in the mounting groove.
上述微流控芯片中设有加样腔体、沉淀腔体和定量腔体,通过加样孔可以向加样腔体中加入待测样本,通过离心分离可以将固体沉淀物与液体分离,得到含有目标物质的待测溶液,加样腔体与第一微流道中的待测溶液可以经毛细流道的毛细力驱动液体流动,最后形成虹吸作用和外界离心作用流至定量腔体中实现待测溶液的定量。该微流控芯片结构相对简单,易于制作成型,可广泛推广使用。The microfluidic chip is equipped with a sample loading cavity, a sedimentation cavity and a quantitative cavity. The sample to be tested can be added to the sample loading cavity through the sample hole, and the solid precipitate can be separated from the liquid by centrifugal separation. For the test solution containing the target substance, the test solution in the sample loading chamber and the first microchannel can drive the liquid to flow through the capillary force of the capillary channel, and finally form a siphon effect and external centrifugal action to flow into the quantitative cavity to achieve the Measure the quantification of the solution. The microfluidic chip has a relatively simple structure, is easy to manufacture and shape, and can be widely promoted and used.
该体外检测装置具有检测单元,可以直接对定量腔体中定量的待测溶液进行检测,操作简单,检测效率高。The in-vitro detection device has a detection unit, which can directly detect the quantitative solution to be tested in the quantitative cavity, and has simple operation and high detection efficiency.
附图说明Description of the drawings
图1为本发明一实施例的微流控芯片的正面结构示意图;FIG. 1 is a schematic diagram of the front structure of a microfluidic chip according to an embodiment of the present invention;
图2为图1所示微流控芯片的背面结构示意图;2 is a schematic diagram of the backside structure of the microfluidic chip shown in FIG. 1;
图3为图1所示微流控芯片配合检测单元的侧视图;Fig. 3 is a side view of the microfluidic chip shown in Fig. 1 with a detection unit;
图4-1、4-2、4-3和4-4为图1所示微流控芯片对全血样本进行血浆(或血清)分离、定量的过程示意图,图4-2-1、4-3-1和4-4-1为相应的局部放大示意图;Figures 4-1, 4-2, 4-3 and 4-4 are schematic diagrams of the process of plasma (or serum) separation and quantification of the whole blood sample by the microfluidic chip shown in Figure 1, Figure 4-2-1, 4 -3-1 and 4-4-1 are corresponding partial enlarged schematic diagrams;
图5-1、5-2和5-3为本发明一实施例的体外检测装置的血浆(或血清)溶解水溶性膜进入检测单元的过程示意图;Figures 5-1, 5-2, and 5-3 are schematic diagrams of the process of dissolving the water-soluble membrane into the detection unit by plasma (or serum) of an in vitro detection device according to an embodiment of the present invention;
图6为一实施例的干化学试纸的结构示意图。FIG. 6 is a schematic diagram of the structure of a dry chemical test paper according to an embodiment.
具体实施方式detailed description
为了便于理解本发明,下面将参照相关附图对本发明进行更全面的描述。附图中给出了本发明的较佳实施例。但是,本发明可以以许多不同的形式来实现,并不限于本文所描述的实施例。相反地,提供这些实施例的目的是使对本发明的公开内容的理解更加透彻全面。In order to facilitate the understanding of the present invention, the present invention will be described more fully below with reference to the relevant drawings. The preferred embodiments of the invention are shown in the drawings. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive.
需要说明的是,当元件被称为“设于”、“安装于”另一个元件,它可以直接在另一个元件上或者也可以存在居中的元件。当一个元件被认为是“连接”另一个元件,它可以是直接连接到另一个元件或者可能同时存在居中元件。It should be noted that when a component is referred to as being "disposed on" or "mounted on" another component, it can be directly on the other component or a centered component may also exist. When an element is considered to be "connected" to another element, it can be directly connected to the other element or an intermediate element may be present at the same time.
除非另有定义,本文所使用的所有的技术和科学术语与属于本发明的技术领域的技术人员通常理解的含义相同。本文中在本发明的说明书中所使用的术语只是为了描述具体的实施例的目的,不是旨在于限制本发明。本文所使用的术语“和/或”包括一个或多个相关的所列项目的任意的和所有的组合。Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the technical field of the present invention. The terms used in the description of the present invention herein are only for the purpose of describing specific embodiments, and are not intended to limit the present invention. The term "and/or" as used herein includes any and all combinations of one or more related listed items.
请结合图1和图2,本发明一实施例提供了一种微流控芯片10,其上设有分离定量单元100。分离定量单元100包括加样腔体110、第一微流道120、沉淀腔体130、毛细流道140、第二微流道150和定量腔体160。加样腔体110用于盛放待测样本,其设有加样孔111。沉淀腔体130用于收集固体等密度较大的待测样本中的无用物。加样腔体110与沉淀腔体130通过第一微流道120连通。第一微流道120通过毛细流道140与第二微流道150连通。第二微流道150与定量腔体160连通。定量腔体160用于定量待测样本溶液。1 and FIG. 2, an embodiment of the present invention provides a microfluidic chip 10 on which a separation and quantification unit 100 is provided. The separation and quantification unit 100 includes a sample addition cavity 110, a first micro flow channel 120, a sedimentation cavity 130, a capillary flow channel 140, a second micro flow channel 150 and a quantification cavity 160. The sample adding cavity 110 is used to hold the sample to be tested, and is provided with a sample adding hole 111. The sedimentation chamber 130 is used to collect useless objects in the sample to be tested with a relatively high density of solids. The sample loading cavity 110 and the sedimentation cavity 130 are in communication with each other through the first micro flow channel 120. The first micro channel 120 communicates with the second micro channel 150 through the capillary channel 140. The second micro flow channel 150 is in communication with the quantitative cavity 160. The quantitative cavity 160 is used to quantify the sample solution to be tested.
在本实施例中,微流控芯片10具有旋转中心18。沉淀腔体130较加样腔体 110远离该旋转中心18。毛细流道140自与第一微流道120连接后向靠近旋转中心18的方向(可以是各逐渐靠近旋转中心18的方向,例如可以是但不限于径向)延伸并弯曲后向远离旋转中心18的方向(可以是各逐渐远离旋转中心18的方向,例如可以是但不限于径向)延伸以与第二微流道150连接。定量腔体160较毛细流道140远离旋转中心。In this embodiment, the microfluidic chip 10 has a center of rotation 18. The sedimentation cavity 130 is farther away from the rotation center 18 than the sample addition cavity 110. The capillary flow channel 140 extends from the connection with the first micro flow channel 120 in the direction approaching the rotation center 18 (may be each gradually approaching the rotation center 18, such as but not limited to the radial direction), and then bends away from the rotation center. The direction of 18 (may be a direction gradually away from the rotation center 18, for example, but not limited to a radial direction) extends to connect with the second micro channel 150. The quantitative cavity 160 is farther from the center of rotation than the capillary flow channel 140.
在图示的具体示例中,加样腔体110与第一微流道120连接的一端的底部倾斜设置,以便于待测样本流至第一微流道120中。In the specific example shown in the figure, the bottom of the end connecting the sample adding cavity 110 and the first micro flow channel 120 is inclined to facilitate the flow of the sample to be tested into the first micro flow channel 120.
第一微流道120的尺寸需满足能够使待测样本中的密度较大的无用物通过,例如对于全血样本,在分离血浆或血清时,第一微流道120需要满足使全血中的血细胞能够通过。在图示的具体示例中,第一微流道120具有向靠近旋转中心18的方向延伸的分支部121,毛细流道140与第一微流道120的该分支部121的末端连接。The size of the first microfluidic channel 120 needs to be able to pass through the useless objects in the sample to be tested. For example, for a whole blood sample, when separating plasma or serum, the first microfluidic channel 120 needs to satisfy Of blood cells can pass through. In the specific example shown in the figure, the first micro flow channel 120 has a branch portion 121 extending in a direction close to the rotation center 18, and the capillary flow channel 140 is connected to the end of the branch portion 121 of the first micro flow channel 120.
在一个具体示例中,该分离定量单元100还包括废液腔体170。废液腔体170用于收集多余的待测溶液。废液腔体170与第二微流道150连通。废液腔体170在第二微流道150上位于定量腔体160的下游,以接收多余的待测溶液。废液腔体170较毛细流道140远离旋转中心18。当废液腔体170中有待测溶液出现时,说明位于其上游的定量腔体160中已装满待测溶液,从而定量腔体160可以实现对待测溶液的定量。In a specific example, the separation and quantitative unit 100 further includes a waste liquid cavity 170. The waste liquid chamber 170 is used to collect excess solution to be tested. The waste liquid cavity 170 is in communication with the second micro flow channel 150. The waste liquid cavity 170 is located downstream of the quantitative cavity 160 on the second micro flow channel 150 to receive the excess solution to be tested. The waste liquid cavity 170 is farther away from the rotation center 18 than the capillary flow channel 140. When there is a solution to be tested in the waste liquid cavity 170, it means that the quantitative cavity 160 located upstream of it is filled with the solution to be tested, so that the quantitative cavity 160 can quantify the solution to be tested.
在一个具体示例中,加样腔体110还设有第一透气孔112。第一透气孔112用于透气,这样可以便于样本的加入,避免因加样时造成腔体内部气压上升而影响样本进入。In a specific example, the sample adding cavity 110 is further provided with a first vent 112. The first vent hole 112 is used for ventilation, which can facilitate the addition of the sample, and avoid the influence of the sample entry due to the increase of the air pressure inside the cavity when the sample is added.
进一步,在一个具体示例中,加样腔体110内在加样孔111与第一透气孔 112之间设有阻流板113。阻流板113在样本加入后可以起到阻挡样本到达第一透气孔112的一侧,防止样本从第一透气孔112流出。Further, in a specific example, a baffle 113 is provided between the sample adding hole 111 and the first vent hole 112 in the sample adding cavity 110. The baffle 113 can prevent the sample from reaching one side of the first vent hole 112 after the sample is added, and prevent the sample from flowing out of the first vent hole 112.
优选地,加样孔111及第一透气孔112在加样腔体110上均靠近旋转中心18设置,这样在转动离心使样本向加样腔体110的远离旋转中心的一侧流动时,可放置样本从加样孔111和第一透气孔112流出,使样本顺利流至第一微流道120和沉淀腔体130中。Preferably, the sample addition hole 111 and the first vent hole 112 are both arranged on the sample addition cavity 110 close to the rotation center 18, so that when the sample is rotated and centrifuged to make the sample flow to the side of the sample addition cavity 110 far from the rotation center, The sample is placed to flow out from the sample loading hole 111 and the first vent hole 112, so that the sample flows smoothly into the first micro flow channel 120 and the sedimentation cavity 130.
毛细流道140呈V字形状,其弯曲部分靠近于旋转中心18。在一个具体示例中,毛细流道140的宽度为0.1mm~0.2mm,深度为0.1mm~0.2mm;或者毛细流道140的宽度为0.2mm~0.5mm,深度为0.2mm~0.5mm。当毛细流道140的宽度为0.1mm~0.2mm,深度为0.1mm~0.2mm时无需进行表面处理,当毛细流道140的宽度为0.2mm~0.5mm,深度为0.2mm~0.5mm时,毛细流道140的流道壁优选经PEG4000表面处理。进一步优选地,毛细流道140的宽度为0.2mm,深度也为0.2mm。毛细流道140在样本溶液进入后,使样本溶液可以借由毛细作用流动至其另一端,并最终在第一微流道120与第二微流道150之间形成虹吸作用。The capillary flow channel 140 has a V shape, and its curved part is close to the rotation center 18. In a specific example, the capillary flow channel 140 has a width of 0.1 mm to 0.2 mm and a depth of 0.1 mm to 0.2 mm; or the capillary flow channel 140 has a width of 0.2 mm to 0.5 mm and a depth of 0.2 mm to 0.5 mm. When the capillary channel 140 has a width of 0.1mm~0.2mm and a depth of 0.1mm~0.2mm, no surface treatment is required. When the capillary channel 140 has a width of 0.2mm~0.5mm and a depth of 0.2mm~0.5mm, The flow channel wall of the capillary flow channel 140 is preferably surface-treated with PEG4000. Further preferably, the width of the capillary flow channel 140 is 0.2 mm, and the depth is also 0.2 mm. After the capillary channel 140 enters the sample solution, the sample solution can flow to the other end of the sample solution by capillary action, and finally form a siphon effect between the first micro channel 120 and the second micro channel 150.
所述PEG4000表面处理可以是但不限于将1wt%的PEG4000溶液加入到毛细流道140中,自然干燥后形成。PEG4000表面处理有利于增加毛细流道140的毛细作用力,并且PEG4000在反应体系中属于惰性物质,一般不会与样本和检测试剂等起反应,因而不会影响检测结果。The PEG4000 surface treatment can be, but is not limited to, adding a 1 wt% PEG4000 solution to the capillary flow channel 140 and forming it after natural drying. The surface treatment of PEG4000 is beneficial to increase the capillary force of the capillary channel 140, and PEG4000 is an inert substance in the reaction system, and generally does not react with samples and detection reagents, and thus does not affect the detection results.
在一个具体示例中,第二微流道150上设有第二透气孔151。第二透气孔151位于第二微流道150上连接的腔体结构(例如定量腔体160和废液腔体170)的下游,且第二透气孔151相对于第二微流道150上连接的腔体结构更靠近于 旋转中心18。第二透气孔151也起到透气的作用,便于待测样本溶液顺利流入第二微流道150,并最终流至定量腔体160和废液腔体170中。In a specific example, the second micro flow channel 150 is provided with a second vent hole 151. The second vent hole 151 is located downstream of the cavity structure (for example, the quantitative cavity 160 and the waste liquid cavity 170) connected to the second micro flow channel 150, and the second vent hole 151 is connected to the second micro flow channel 150 The cavity structure is closer to the center of rotation 18. The second vent hole 151 also plays a role of ventilating, facilitating the sample solution to be tested to smoothly flow into the second micro flow channel 150 and finally flow into the quantitative cavity 160 and the waste liquid cavity 170.
可理解,在其他具体示例中,第一透气孔112和第二透气孔151可以择一,例如可以只有第一透气孔112,也可以只有第二透气孔151,其中,第二透气孔151为择一之优选。It can be understood that in other specific examples, the first vent 112 and the second vent 151 can be selected. For example, there may be only the first vent 112 or the second vent 151, wherein the second vent 151 is Choose one of the best.
进一步,第二微流道150上位于其连接的腔体结构的下游的部分向靠近旋转中心18的方向弯折延伸,第二透气孔151设于第二微流道150的末端,这样可以有效防止样本溶液从第二透气孔151流出。Further, the part of the second micro-channel 150 downstream of the cavity structure connected to it is bent and extended in a direction close to the rotation center 18, and the second vent hole 151 is provided at the end of the second micro-channel 150, which can effectively The sample solution is prevented from flowing out from the second vent hole 151.
请参图3,在一个具体示例中,微流控芯片10包括芯片本体11和覆盖在芯片本体11上的透明盖膜12。芯片本体11与透明盖膜12配合形成分离定量单元100的各腔体结构和流道结构。具体地,各腔体结构和流道结构的沟槽等均预形成在芯片本体11上,如图2所示,各孔均开口在芯片本体的背面,后续通过透明盖膜12覆盖并密封在芯片本体11的正面即可形成完成对腔体结构和流道结构的封装,形成完整的腔体结构和流道结构。Please refer to FIG. 3, in a specific example, the microfluidic chip 10 includes a chip body 11 and a transparent cover film 12 covering the chip body 11. The chip body 11 and the transparent cover film 12 cooperate to form the cavity structure and the flow channel structure of the separation and quantitative unit 100. Specifically, the grooves of each cavity structure and flow channel structure are pre-formed on the chip body 11. As shown in FIG. 2, each hole is opened on the back of the chip body, and is subsequently covered by a transparent cover film 12 and sealed in The front surface of the chip body 11 can be formed to complete the packaging of the cavity structure and the flow channel structure, forming a complete cavity structure and the flow channel structure.
透明盖膜12可以是但不限于透明胶带或者透明压敏胶等,其与芯片本体11配合构成整个微流控芯片10,装配简单,无需使用复杂、昂贵的超声焊接技术,直接粘接即可,可以显著降低制作成本。可理解,在其他具体示例中,微流控芯片10也可以采用成本较高的超声焊接技术焊接形成,或者采用3D打印技术一体成型。The transparent cover film 12 can be, but is not limited to, transparent tape or transparent pressure-sensitive adhesive, etc., which cooperates with the chip body 11 to form the entire microfluidic chip 10, which is simple to assemble and does not require complicated and expensive ultrasonic welding technology. , Can significantly reduce production costs. It can be understood that, in other specific examples, the microfluidic chip 10 may also be formed by welding with a relatively high-cost ultrasonic welding technology, or be integrally formed with a 3D printing technology.
在一个具体示例中,分离定量单元100有多个,多个分离定量单元100围绕同一旋转中心18设置。通过设置多个定量单元100可以实现多样本单项检测,也可以实现单样本多项检测,一致性好,集成度高,显著提高了单次检测的通 量。In a specific example, there are multiple separation and quantitative units 100, and the multiple separation and quantitative units 100 are arranged around the same rotation center 18. Multiple quantification units 100 can realize multi-sample single detection, and can also realize single-sample multiple detection, with good consistency and high integration, which significantly improves the throughput of single detection.
优选地,例如在图示的具体示例中,微流控芯片10呈圆盘状,多个分离定量单元100均匀分布在微流控芯片10上,微流控芯片10的中部具有旋转安装部180,旋转安装部180的中心即旋转中心18。旋转安装部180可以是各类卡槽或者卡柱等结构。Preferably, for example, in the specific example shown in the figure, the microfluidic chip 10 is in the shape of a disc, a plurality of separate quantitative units 100 are evenly distributed on the microfluidic chip 10, and the microfluidic chip 10 has a rotating mounting part 180 in the middle. , The center of the rotating mounting portion 180 is the center of rotation 18. The rotating mounting part 180 may be various types of clamping slots or clamping posts.
上述微流控芯片10中设有加样腔体110、沉淀腔体130和定量腔体160,通过加样孔111可以向加样腔体110中加入待测样本,通过离心分离可以将固体沉淀物与液体分离,得到含有目标物质的待测溶液,加样腔体110与第一微流道120中的待测溶液可以经毛细流道140的毛细力驱动液体流动,最后形成虹吸作用和外界离心作用流至定量腔体160中实现对待测溶液的定量。该微流控芯片10结构相对简单,易于制作成型,可广泛推广使用。The microfluidic chip 10 is provided with a sample loading cavity 110, a precipitation cavity 130, and a quantitative cavity 160. The sample to be tested can be added to the sample loading cavity 110 through the sample hole 111, and the solid can be precipitated by centrifugal separation. The substance is separated from the liquid to obtain the test solution containing the target substance. The test solution in the sample loading chamber 110 and the first microchannel 120 can drive the liquid to flow through the capillary force of the capillary channel 140, and finally form a siphon effect and the outside world. The centrifugal action flows into the quantification cavity 160 to realize the quantification of the solution to be tested. The microfluidic chip 10 has a relatively simple structure, is easy to manufacture and shape, and can be widely promoted and used.
请参图2、3、图5-1、5-2和5-3,本发明还提供了一种体外检测装置2,其包括上述微流控芯片10和检测单元20。检测单元20用于检测微流控芯片10的定量腔体160内的样本。Referring to FIGS. 2, 3, FIGS. 5-1, 5-2 and 5-3, the present invention also provides an in vitro detection device 2, which includes the aforementioned microfluidic chip 10 and a detection unit 20. The detection unit 20 is used to detect the sample in the quantitative cavity 160 of the microfluidic chip 10.
在一个具体示例中,检测单元20为设置在定量腔体160内的冻干试剂。冻干试剂置于定量腔体160内,当样本溶液流至定量腔体160中时,即可溶解该冻干试剂,并与之反应,对反应的结果进行检测即可。In a specific example, the detection unit 20 is a freeze-dried reagent arranged in the quantitative cavity 160. The freeze-dried reagent is placed in the quantitative cavity 160. When the sample solution flows into the quantitative cavity 160, the freeze-dried reagent can be dissolved and reacted with it, and the result of the reaction can be detected.
由于冻干试剂制作过程复杂,成本较高,在另一个具体示例中,检测单元20外置。具体地,定量腔体160具有渗透孔161,渗透孔161上覆盖有水溶性膜162。检测单元20的进样口21与渗透孔161对接且由水溶性膜162隔开。Due to the complicated production process of the freeze-dried reagents and the high cost, in another specific example, the detection unit 20 is external. Specifically, the quantitative cavity 160 has a permeation hole 161, and the permeation hole 161 is covered with a water-soluble film 162. The injection port 21 of the detection unit 20 is connected to the permeation hole 161 and is separated by a water-soluble membrane 162.
进一步,在一个具体示例中,检测单元20为干化学试纸。如图6所示,该干化学试纸20包括支撑层22和在支撑层22上依次层叠设置的反应指示层23 和扩散层24。反应指示层23中含有能够与待测样本中目标物质反应的反应试剂和指示试剂。反应指示层23可以是一层,也可以是多层,例如在图6所示的具体示例中,该反应指示层23包括指示层231和试剂层232两层,指示层231靠近于支撑层22,其中含有显色指示试剂,试剂层232靠近于扩散层24,其中含有能够与目标物质反应的反应试剂;此外,指示层232与试剂层232中所含的试剂也可以对换或适当混合。扩散层24通过进样口21面向于水溶性膜162。Further, in a specific example, the detection unit 20 is a dry chemical test paper. As shown in FIG. 6, the dry chemical test paper 20 includes a supporting layer 22 and a reaction indicating layer 23 and a diffusion layer 24 stacked on the supporting layer 22 in sequence. The reaction indicator layer 23 contains a reaction reagent and an indicator reagent that can react with the target substance in the sample to be tested. The reaction indicating layer 23 may be one layer or multiple layers. For example, in the specific example shown in FIG. 6, the reaction indicating layer 23 includes two layers of an indicating layer 231 and a reagent layer 232, and the indicating layer 231 is close to the supporting layer 22. , Which contains a color indicator reagent, the reagent layer 232 is close to the diffusion layer 24, and contains a reaction reagent capable of reacting with the target substance; in addition, the reagents contained in the indicator layer 232 and the reagent layer 232 can also be exchanged or appropriately mixed. The diffusion layer 24 faces the water-soluble film 162 through the injection port 21.
更进一步,微流控芯片10围绕渗透孔设有安装槽。检测单元20镶嵌在安装槽中。Furthermore, the microfluidic chip 10 is provided with a mounting groove around the penetration hole. The detection unit 20 is embedded in the installation groove.
该体外检测装置2具有检测单元20,可以直接对定量腔体160中定量的待测样本溶液进行检测,操作简单,检测效率高。以全血样本上样检测为例,使用干化学试纸的体外检测装置2的具体检测过程可参考如下:The in vitro detection device 2 has a detection unit 20, which can directly detect the quantified sample solution to be tested in the quantitative cavity 160, with simple operation and high detection efficiency. Taking the whole blood sample loading test as an example, the specific test process of the in vitro test device 2 using dry chemical test paper can be referred to as follows:
请参图4-1、4-2、4-3和4-4,将一定量的全血通过加样孔111加入至加样腔体110中,依次可以加入六人份不同的样本;Please refer to Figures 4-1, 4-2, 4-3 and 4-4, and add a certain amount of whole blood to the sample chamber 110 through the sample hole 111, and six different samples can be added in sequence;
加完样本后,将体外检测装置2的旋转安装部180安装于配套的旋转离心仪器中,开启仪器转动,全血在离心力的作用下,红细胞等沉淀在沉淀腔体130中,血清或血浆分离至沉淀腔体130的上部、第二微流道120以及加样腔体110中;After the sample is added, install the rotary mounting part 180 of the in vitro detection device 2 in the supporting rotary centrifugal instrument, turn on the instrument to rotate, the whole blood is subjected to centrifugal force, and the red blood cells are deposited in the sedimentation chamber 130, and the serum or plasma is separated To the upper part of the precipitation chamber 130, the second micro flow channel 120, and the sample adding chamber 110;
微流控芯片10在转动时,血清或血浆只是部分填充在毛细流道140中,如图4-2-1和4-3-1所示,其液位超过毛细流道140的入口端140a,而不超过毛细流道140的弯曲部位140b;当微流控芯片10停止转动时,在毛细流道140的毛细力作用下,血清或血浆越过弯曲部位140b,到达毛细流道140的末端140c,由于末端140c的点低于加样腔体110中的液位高度(较加样腔体110中的液位 远离旋转中心18),因而可以形成虹吸作用;When the microfluidic chip 10 rotates, serum or plasma is only partially filled in the capillary channel 140, as shown in Figures 4-2-1 and 4-3-1, the liquid level exceeds the inlet end 140a of the capillary channel 140 , Without exceeding the curved part 140b of the capillary flow channel 140; when the microfluidic chip 10 stops rotating, under the capillary force of the capillary flow channel 140, the serum or plasma crosses the curved part 140b and reaches the end 140c of the capillary flow channel 140 , Since the point at the end 140c is lower than the height of the liquid level in the sample loading cavity 110 (which is farther from the rotation center 18 than the liquid level in the sample loading cavity 110), a siphon effect can be formed;
当毛细流道140中填充满血清或血浆后,控制微流控芯片10再次旋转,由于末端140c较加样腔体110中的液位远离旋转中心18,如图4-3-1和4-4-1所示,在虹吸作用和离心力的作用下,血清或血浆通过第二微流道150进入定量腔体160,当定量腔体160中充满血清或血浆后,即完成对待测样本溶液的定量,多余的血清或血浆进入第二微流道150的后段或者进入废液腔体170中;优选地,仪器会根据第二微流道150的后段或者废液腔体170中是否有液体判断定量腔体160是否充满液体,当检测到第二微流道150的后段或者废液腔体170中有液体时,即可确定定量腔体160充满液体,否则报警提醒需要再次检测定量腔体160是否充满液体;When the capillary channel 140 is filled with serum or plasma, the microfluidic chip 10 is controlled to rotate again, because the end 140c is farther from the center of rotation 18 than the liquid level in the sample cavity 110, as shown in Figures 4-3-1 and 4- As shown in 4-1, under the action of siphoning and centrifugal force, serum or plasma enters the quantitative cavity 160 through the second microchannel 150. When the quantitative cavity 160 is filled with serum or plasma, the sample solution to be tested is completed Quantitatively, the excess serum or plasma enters the back section of the second micro-channel 150 or the waste liquid cavity 170; preferably, the instrument will determine whether there is in the back section of the second micro-channel 150 or the waste liquid cavity 170. The liquid judges whether the quantitative cavity 160 is full of liquid. When it is detected that there is liquid in the back section of the second microfluidic channel 150 or the waste liquid cavity 170, it can be determined that the quantitative cavity 160 is full of liquid. Whether the cavity 160 is full of liquid;
从血清或血浆充满毛细流道140时开启转动至定量腔体160中充满液体,一般在10s内即可完成,当定量腔体160中充满液体时,控制微流控芯片10停止转动,静置,液体会暂时封闭在定量腔体160中,随着时间推移,液体会逐渐将渗透孔161处覆盖的水溶性膜162溶解,该溶解过程大约需要1min,如图5-1、5-2和5-3所示;Turn on and turn when the capillary channel 140 is filled with serum or plasma until the quantitative cavity 160 is filled with liquid, generally within 10s. When the quantitative cavity 160 is filled with liquid, control the microfluidic chip 10 to stop rotating and stand still , The liquid will be temporarily enclosed in the quantitative cavity 160. As time goes by, the liquid will gradually dissolve the water-soluble film 162 covered at the permeation hole 161. The dissolution process takes about 1 min, as shown in Figures 5-1, 5-2 and Shown in 5-3;
当水溶性膜162溶解掉之后,液体一般不会因为自重而流入检测单元(干化学试纸)20,需要控制转动微流控芯片10,可以在低速1800rpm~2000rpm的速度下转动,使定量腔体160中定量的样本溶液进入检测单元20的进样口21;After the water-soluble film 162 is dissolved, the liquid generally does not flow into the detection unit (dry chemical test paper) 20 due to its own weight. The microfluidic chip 10 needs to be rotated under control. It can rotate at a low speed of 1800rpm~2000rpm to make the quantitative chamber The quantified sample solution in 160 enters the sample inlet 21 of the detection unit 20;
样本溶液依次经扩散层24扩散至反应指示层23进行显色反应,显色的深浅可反映出检测物的浓度,通过检测单元的检测孔25可进行信号采集,最后转化为检测物的浓度数据。该体外检测装置2可以解决干化学试纸全血灵敏度和稳定性低的问题,具有检测通量高、成本低等优点。The sample solution is sequentially diffused through the diffusion layer 24 to the reaction indicator layer 23 for color reaction. The depth of the color can reflect the concentration of the test substance. The signal can be collected through the detection hole 25 of the detection unit, and finally converted into the concentration data of the test substance. . The in vitro detection device 2 can solve the problems of low sensitivity and stability of dry chemical test paper whole blood, and has the advantages of high detection throughput and low cost.
以上所述实施例的各技术特征可以进行任意的组合,为使描述简洁,未对上述实施例中的各个技术特征所有可能的组合都进行描述,然而,只要这些技术特征的组合不存在矛盾,都应当认为是本说明书记载的范围。The technical features of the above-mentioned embodiments can be combined arbitrarily. In order to make the description concise, all possible combinations of the technical features in the above-mentioned embodiments are not described. However, as long as there is no contradiction in the combination of these technical features, All should be considered as the scope of this specification.
以上所述实施例仅表达了本发明的几种实施方式,其描述较为具体和详细,但并不能因此而理解为对发明专利范围的限制。应当指出的是,对于本领域的普通技术人员来说,在不脱离本发明构思的前提下,还可以做出若干变形和改进,这些都属于本发明的保护范围。因此,本发明专利的保护范围应以所附权利要求为准。The above-mentioned embodiments only express several embodiments of the present invention, and the descriptions are more specific and detailed, but they should not be understood as limiting the scope of the invention patent. It should be pointed out that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can be made, and these all fall within the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention should be subject to the appended claims.

Claims (17)

  1. 一种微流控芯片,其特征在于,所述微流控芯片上设有分离定量单元,所述分离定量单元包括加样腔体、第一微流道、沉淀腔体、毛细流道、第二微流道和定量腔体;所述加样腔体设有加样孔,所述加样腔体与所述沉淀腔体通过所述第一微流道连通,所述第一微流道通过所述毛细流道与所述第二微流道连通,所述第二微流道与所述定量腔体连通;A microfluidic chip, characterized in that a separation and quantification unit is provided on the microfluidic chip, and the separation and quantification unit includes a sample addition chamber, a first micro flow channel, a precipitation chamber, a capillary flow channel, and a second Two micro flow channels and a quantitative cavity; the sample loading cavity is provided with a sample loading hole, the sample loading cavity and the precipitation cavity are communicated through the first micro flow channel, and the first micro flow channel Communicate with the second micro flow channel through the capillary flow channel, and the second micro flow channel communicates with the quantitative cavity;
    所述微流控芯片具有旋转中心,所述沉淀腔体较所述加样腔体远离所述旋转中心,所述毛细流道自与所述第一微流道连接后向靠近所述旋转中心的方向延伸并弯曲后向远离所述旋转中心的方向延伸以与所述第二微流道连接,所述定量腔体较所述毛细流道远离所述旋转中心。The microfluidic chip has a center of rotation, the precipitation chamber is farther from the center of rotation than the sample loading chamber, and the capillary channel is closer to the center of rotation after being connected to the first microchannel. It extends in the direction of and bends and then extends in a direction away from the rotation center to connect with the second micro flow channel, and the quantitative cavity is farther from the rotation center than the capillary flow channel.
  2. 如权利要求1所述的微流控芯片,其特征在于,所述分离定量单元还包括废液腔体,所述废液腔体与所述第二微流道连通,所述废液腔体在所述第二微流道上位于所述定量腔体的下游,所述废液腔体较所述毛细流道远离所述旋转中心。The microfluidic chip according to claim 1, wherein the separation and quantification unit further comprises a waste liquid cavity, the waste liquid cavity is in communication with the second microfluidic channel, and the waste liquid cavity The second micro flow channel is located downstream of the quantitative cavity, and the waste liquid cavity is farther from the rotation center than the capillary flow channel.
  3. 如权利要求1所述的微流控芯片,其特征在于,所述加样腔体还设有第一透气孔。3. The microfluidic chip of claim 1, wherein the sample loading cavity is further provided with a first vent.
  4. 如权利要求3所述的微流控芯片,其特征在于,所述加样腔体内在所述加样孔与所述第一透气孔之间设有阻流板。8. The microfluidic chip of claim 3, wherein a baffle plate is provided in the sample loading cavity between the sample loading hole and the first vent hole.
  5. 如权利要求3所述的微流控芯片,其特征在于,所述加样孔及所述第一透气孔在所述加样腔体上均靠近所述旋转中心设置。8. The microfluidic chip of claim 3, wherein the sample application hole and the first vent hole are both arranged on the sample application cavity close to the rotation center.
  6. 如权利要求1所述的微流控芯片,其特征在于,所述毛细流道的宽度为0.1mm~0.2mm,深度为0.1mm~0.2mm;或者The microfluidic chip of claim 1, wherein the capillary flow channel has a width of 0.1 mm to 0.2 mm and a depth of 0.1 mm to 0.2 mm; or
    所述毛细流道的宽度为0.2mm~0.5mm,深度为0.2mm~0.5mm,所述毛细流道的流道壁经PEG4000表面处理。The width of the capillary flow channel is 0.2mm-0.5mm, and the depth is 0.2mm-0.5mm, and the flow channel wall of the capillary flow channel is surface treated with PEG4000.
  7. 如权利要求1所述的微流控芯片,其特征在于,所述第二微流道上设有第二透气孔,所述第二透气孔位于所述第二微流道上连接的腔体结构的下游,且所述第二透气孔相对于所述第二微流道上连接的腔体结构更靠近于所述旋转中心。The microfluidic chip of claim 1, wherein the second microfluidic channel is provided with a second vent hole, and the second vent hole is located in the cavity structure connected on the second microfluidic channel. Downstream, and the second vent hole is closer to the rotation center than the cavity structure connected to the second micro flow channel.
  8. 如权利要求7所述的微流控芯片,其特征在于,所述第二微流道上位于其连接的腔体结构的下游的部分向靠近所述旋转中心的方向弯折延伸,所述第二透气孔设于所述第二微流道的末端。The microfluidic chip according to claim 7, wherein a part of the second microfluidic channel located downstream of the cavity structure to which it is connected is bent and extends in a direction close to the rotation center, and the second The air vent is provided at the end of the second micro flow channel.
  9. 如权利要求1~8中任一项所述的微流控芯片,其特征在于,所述分离定量单元有多个,多个所述分离定量单元围绕同一所述旋转中心设置。8. The microfluidic chip according to any one of claims 1 to 8, wherein there are multiple separation and quantitative units, and multiple separation and quantitative units are arranged around the same center of rotation.
  10. 如权利要求9所述的微流控芯片,其特征在于,所述微流控芯片呈圆盘状,多个所述分离定量单元均匀分布在所述微流控芯片上,所述微流控芯片的中部具有旋转安装部,所述旋转安装部的中心即所述旋转中心。The microfluidic chip of claim 9, wherein the microfluidic chip is in the shape of a disc, a plurality of the separation and quantification units are evenly distributed on the microfluidic chip, and the microfluidic chip The middle of the chip has a rotating mounting part, and the center of the rotating mounting part is the rotation center.
  11. 如权利要求1~8和10中任一项所述的微流控芯片,其特征在于,所述微流控芯片包括芯片本体和覆盖在所述芯片本体上的透明盖膜,所述芯片本体与所述透明盖膜配合形成所述分离定量单元的各腔体结构和流道结构。The microfluidic chip according to any one of claims 1 to 8 and 10, wherein the microfluidic chip comprises a chip body and a transparent cover film covering the chip body, the chip body Cooperating with the transparent cover film to form each cavity structure and flow channel structure of the separation and quantitative unit.
  12. 一种体外检测装置,其特征在于,包括如权利要求1~11中任一项所述的微流控芯片和检测单元,所述检测单元用于检测所述定量腔体内的样本。An in vitro detection device, characterized by comprising the microfluidic chip according to any one of claims 1 to 11 and a detection unit, the detection unit being used for detecting a sample in the quantitative cavity.
  13. 如权利要求12所述的体外检测装置,其特征在于,所述检测单元为设置在所述定量腔体内的冻干试剂。The in vitro detection device of claim 12, wherein the detection unit is a freeze-dried reagent set in the quantitative cavity.
  14. 如权利要求12所述的体外检测装置,其特征在于,所述定量腔体具有 渗透孔,所述渗透孔上覆盖有水溶性膜,所述检测单元的进样口与所述渗透孔对接且由所述水溶性膜隔开。The in vitro detection device according to claim 12, wherein the quantitative cavity has a permeable hole, the permeable hole is covered with a water-soluble film, and the sampling port of the detection unit is connected to the permeable hole. Separated by the water-soluble membrane.
  15. 如权利要求14所述的体外检测装置,其特征在于,所述检测单元为干化学试纸。The in vitro detection device of claim 14, wherein the detection unit is a dry chemical test paper.
  16. 如权利要求15所述的体外检测装置,其特征在于,所述干化学试纸包括支撑层和在所述支撑层上依次层叠设置的反应指示层和扩散层,所述反应指示层中含有能够与待测样本中目标物质反应的反应试剂和指示试剂,所述扩散层通过所述进样口面向于所述水溶性膜。The in vitro detection device according to claim 15, wherein the dry chemical test paper comprises a support layer and a reaction indicator layer and a diffusion layer stacked on the support layer, and the reaction indicator layer contains The reaction reagent and indicator reagent for the reaction of the target substance in the sample to be tested, and the diffusion layer faces the water-soluble membrane through the sample inlet.
  17. 如权利要求14~16中任一项所述的体外检测装置,其特征在于,所述微流控芯片围绕所述渗透孔设有安装槽,所述检测单元镶嵌在所述安装槽中。The in vitro detection device according to any one of claims 14 to 16, wherein the microfluidic chip is provided with an installation groove around the penetration hole, and the detection unit is embedded in the installation groove.
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