CN106824317A - A kind of method that drop is manipulated using micro- electromagnetic wand - Google Patents
A kind of method that drop is manipulated using micro- electromagnetic wand Download PDFInfo
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
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Abstract
本发明涉及一种利用微电磁棒操控液滴的方法，所述方法基于磁力、表面张力和重力的竞争，以经过超疏水表面处理的微电磁棒和基片为平台，通过开关微电磁棒实现对含磁珠液滴在超疏水基片表面的吸、放操作，同时结合电磁棒的位移或者液滴座落平台的位移，完成对含磁珠液滴的输运、融合、混合操作以及完成含磁珠液滴中固相磁珠与液相的分离操控。本发明提供的利用微电磁棒操控液滴的方法，能够快速、灵活地实现液滴操控，有望促进数字微流控技术的实际应用和快速发展。
The invention relates to a method for manipulating liquid droplets using a micro-electromagnetic rod. The method is based on the competition of magnetism, surface tension and gravity, and uses a micro-electromagnetic rod and a substrate treated with a super-hydrophobic surface as a platform, and is realized by switching the micro-electromagnetic rod. The suction and release operation of the droplet containing magnetic beads on the surface of the superhydrophobic substrate, combined with the displacement of the electromagnetic rod or the displacement of the droplet seating platform, completes the transportation, fusion, mixing operations and completion of the droplet containing magnetic beads Separation and manipulation of solid-phase magnetic beads and liquid phase in droplets containing magnetic beads. The method for manipulating liquid droplets by using a micro-electromagnetic rod provided by the present invention can quickly and flexibly realize liquid droplet manipulation, and is expected to promote the practical application and rapid development of digital microfluidic technology.
Description
技术领域technical field
本发明属于微流控技术领域，涉及一种利用微电磁棒操控液滴的方法。The invention belongs to the technical field of microfluidics, and relates to a method for manipulating liquid droplets by using a micro-electromagnetic rod.
背景技术Background technique
基于液滴操控的数字微流控技术(Digital Microfludics)具有控制单个液滴的能力，可控性好，能耗低，无需复杂的微泵、微阀结构，可应用于高集成度、多步骤复杂操作的微生化分析过程，近年来在微型生物和化学分析领域受到广泛关注，成为当前微流控技术发展最活跃的领域之一。Digital Microfluidics based on droplet manipulation has the ability to control a single droplet, with good controllability, low energy consumption, no need for complex micropumps and microvalve structures, and can be applied to high integration, multi-step The micro-biochemical analysis process with complex operations has received extensive attention in the field of micro-biological and chemical analysis in recent years, and has become one of the most active fields in the development of microfluidic technology.
目前数字微流控技术主要采用电润湿方式(Electrowetting)进行样品液滴的操控，电润湿驱动方式是通过在介质膜下面的微电极阵列上施加电势来改变介质膜与表面液体的浸润特性,即通过局部改变液滴和固体表面的三相接触角，造成液滴两端不对称形变，使液滴内部产生压强差，来实现对液滴传输的操作和控制，包括液滴的取样、输运、混合、分裂等。但是基于电润湿方式数字微流控技术需要在芯片上制作复杂的微电极阵列，工艺加工难度大，芯片成本较高，限制了该类芯片的广泛应用；且该类芯片往往受制于其集成微电极阵列的几何尺寸、排布等因素，在可操控液滴的体积、输运路径和范围等方面受到很大限制，同时基片表面金属电极的存在也影响了透射式光学观测的应用。At present, digital microfluidic technology mainly uses electrowetting to control the sample droplets. The electrowetting driving method is to change the wetting characteristics of the dielectric film and the surface liquid by applying a potential to the microelectrode array under the dielectric film. , That is, by locally changing the three-phase contact angle between the droplet and the solid surface, causing asymmetric deformation at both ends of the droplet, causing a pressure difference inside the droplet to realize the operation and control of droplet transmission, including droplet sampling, Transportation, mixing, splitting, etc. However, digital microfluidic technology based on electrowetting requires complex microelectrode arrays to be fabricated on the chip, which is difficult to process and high in chip cost, which limits the wide application of this type of chip; and this type of chip is often limited by its integration. The geometric size, arrangement and other factors of the microelectrode array are greatly limited in the volume, transport path and range of the droplet that can be manipulated. At the same time, the existence of metal electrodes on the substrate surface also affects the application of transmission optical observation.
虽然近年来也有一些利用光学、热毛细作用和声表面波等技术进行液滴操控的报道，但是这些方法在液体操控的灵活性、应用的可靠性方面均具有较大的局限。而基于磁力的微液滴操控，由于其操作简便，且兼容成熟的磁分离技术，因而近年来受到广泛关注。但是，目前基于磁力的微液滴操控技术，要么采用永磁体，要么采用集成于芯片表面的微磁圈，实现包含磁珠液滴在芯片表面的输运和混合等操控。两种方式均有各自的局限，永磁体方法由于磁体体积相对较大，难以实现多个密集排布磁性液滴中单一目标液滴的高分辨率操控，且难以实现显微光学实时观测；而集成式微磁圈方法则需要复杂昂贵的微加工过程制作芯片，成本较高，不利于广泛应用。因此，需要发展同时具备操作简便、灵活、可控性好、成本低廉、可实现复杂操控的液滴操控平台。Although there have been some reports on liquid droplet manipulation using optics, thermocapillary action, and surface acoustic wave technologies in recent years, these methods have limitations in the flexibility of liquid manipulation and application reliability. Magnetic-based micro-droplet manipulation has attracted extensive attention in recent years due to its ease of operation and compatibility with mature magnetic separation techniques. However, the current micro-droplet manipulation technology based on magnetic force either uses permanent magnets or micro-magnetic coils integrated on the chip surface to realize the transportation and mixing of droplets containing magnetic beads on the chip surface. Both methods have their own limitations. Due to the relatively large volume of the magnet, the permanent magnet method is difficult to achieve high-resolution manipulation of a single target droplet in multiple densely arranged magnetic droplets, and it is difficult to achieve real-time microscopic optical observation; and The integrated micro-magnetic coil method requires a complex and expensive micro-processing process to make chips, and the cost is high, which is not conducive to wide application. Therefore, it is necessary to develop a droplet control platform that is easy to operate, flexible, good controllability, low cost, and capable of complex manipulation.
发明内容Contents of the invention
有鉴于此，本发明的目的在于提供一种利用微电磁棒操控液滴的方法，能够快速、灵活地实现液滴操控，有望促进数字微流控技术的实际应用和快速发展。In view of this, the purpose of the present invention is to provide a method for manipulating liquid droplets using a micro-electromagnetic rod, which can quickly and flexibly realize liquid droplet manipulation, and is expected to promote the practical application and rapid development of digital microfluidic technology.
为达到上述目的，本发明提供如下技术方案：To achieve the above object, the present invention provides the following technical solutions:
一种利用微电磁棒操控液滴的方法，所述方法基于磁力、表面张力和重力的竞争，以经过超疏水表面处理的微电磁棒和基片为平台，通过开关微电磁棒实现对含磁珠液滴在超疏水基片表面的吸、放操作，同时结合电磁棒的位移或者液滴座落平台的位移，完成对含磁珠液滴的输运、融合、混合操作以及完成含磁珠液滴中固相磁珠与液相的分离操控。A method for manipulating liquid droplets using a micro-electromagnetic rod. The method is based on the competition of magnetism, surface tension and gravity. The micro-electromagnetic rod and the substrate have been treated with a super-hydrophobic surface as a platform, and the magnetic field is controlled by switching the micro-electromagnetic rod. The suction and release operation of the bead droplets on the surface of the superhydrophobic substrate, combined with the displacement of the electromagnetic rod or the displacement of the droplet seating platform, completes the transportation, fusion, and mixing of the droplets containing magnetic beads Separation and manipulation of solid-phase magnetic beads and liquid phase in droplets.
进一步地，所用微电磁棒由磁芯微棒缠绕通电线圈制成；所述磁芯微棒的截面为圆形，所述磁芯微棒与含磁珠液滴接触的一端为锥形端，且所述锥形端预先经超疏水表面处理。Further, the micro-rod used is made of a magnetic core microrod wound with an energized coil; the cross section of the magnetic core microrod is circular, and the end of the magnetic core microrod in contact with the droplet containing magnetic beads is a tapered end, And the tapered end is pre-treated with a super-hydrophobic surface.
进一步地，所用磁芯的材料为铁、锰-锌铁氧体或镍-锌铁氧体，所述磁芯微棒的直径范围为1～10mm。Further, the material of the magnetic core used is iron, manganese-zinc ferrite or nickel-zinc ferrite, and the diameter of the magnetic core microrod ranges from 1 to 10 mm.
进一步地，所述磁珠为顺磁性微球，直径范围为1～100μm。Further, the magnetic beads are paramagnetic microspheres with a diameter ranging from 1 to 100 μm.
进一步地，所述超疏水基片的表面包含凹形局域结构或者亲水局域结构。Further, the surface of the superhydrophobic substrate includes a concave localized structure or a hydrophilic localized structure.
进一步地，所述含磁珠液滴的输运操作步骤如下：Further, the transport operation steps of the droplet containing magnetic beads are as follows:
将所述微电磁棒的尖端接触位于所述超疏水基片表面的含磁珠液滴；The tip of the micro-electromagnetic rod is contacted with the droplet containing magnetic beads on the surface of the superhydrophobic substrate;
开通所述微电磁棒的电源，使其尖端产生磁场，通过磁力作用吸住所述含磁珠液滴中的磁珠；Turn on the power supply of the micro-electromagnetic rod, make its tip generate a magnetic field, and absorb the magnetic beads in the droplet containing magnetic beads through magnetic force;
上移所述微电磁棒，使所述微电磁棒利用吸住的磁珠通过表面张力提起所述含磁珠液滴；Moving up the micro-electromagnetic rod, so that the micro-electromagnetic rod utilizes the magnetic beads absorbed to lift the droplet containing magnetic beads through surface tension;
平移携带所述含磁珠液滴的微电磁棒至目标位置上方，下移所述微电磁棒，使所述含磁珠液滴接触目标位置基片表面；Translating the micro-electromagnetic rod carrying the magnetic bead-containing droplet above the target position, and moving down the micro-electromagnetic rod so that the magnetic bead-containing droplet contacts the surface of the substrate at the target position;
关闭所述微电磁棒的电源，消除磁力作用，使所述含磁珠液滴与所述微电磁棒脱离，在重力作用下坐落于目标位置，完成液滴输运。Turning off the power supply of the micro-electromagnetic rod, eliminating the magnetic force, so that the droplet containing magnetic beads is separated from the micro-electromagnetic rod, and is located at the target position under the action of gravity to complete the droplet transportation.
进一步地，所述含磁珠液滴的融合操作步骤如下：Further, the fusion operation steps of the droplet containing magnetic beads are as follows:
将所述微电磁棒的尖端接触位于所述超疏水基片表面的含磁珠液滴；The tip of the micro-electromagnetic rod is contacted with the droplet containing magnetic beads on the surface of the superhydrophobic substrate;
开通所述微电磁棒的电源，使其尖端产生磁场，通过磁力作用吸住所述含磁珠液滴中的磁珠；Turn on the power supply of the micro-electromagnetic rod, make its tip generate a magnetic field, and absorb the magnetic beads in the droplet containing magnetic beads through magnetic force;
上移所述微电磁棒，使所述微电磁棒利用吸住的磁珠通过表面张力提起所述含磁珠液滴；Moving up the micro-electromagnetic rod, so that the micro-electromagnetic rod utilizes the magnetic beads absorbed to lift the droplet containing magnetic beads through surface tension;
平移携带所述含磁珠液滴的微电磁棒至目标液滴上方，下移微电磁棒，使所述含磁珠液滴与目标液滴接触，通过表面张力作用实现融合；Translating the micro-electromagnetic rod carrying the magnetic bead-containing droplet above the target droplet, moving the micro-electromagnetic rod downward to make the magnetic-bead-containing droplet contact the target droplet, and achieve fusion through surface tension;
关闭所述微电磁棒的电源，消除磁力作用，使融合的液滴与微电磁棒脱离，完成液滴融合。Turn off the power supply of the micro-electromagnetic rod, eliminate the magnetic force, and separate the fused droplets from the micro-electromagnetic rod to complete the droplet fusion.
进一步地，所述含磁珠液滴的混合操作步骤如下：Further, the mixing operation steps of the magnetic bead-containing droplets are as follows:
将微电磁棒尖端接触包含磁珠的融合液滴；Touch the tip of the microelectromagnetic rod to the fused droplet containing the magnetic beads;
开通微电磁棒电源，使其尖端产生磁场，通过磁力作用吸住所述融合液滴中的磁珠；Turn on the power supply of the micro-electromagnetic rod, make its tip generate a magnetic field, and absorb the magnetic beads in the fusion droplet through magnetic force;
使微电磁棒沿所述融合液滴表面往复运动或绕行，带动所述融合液滴中的磁珠相对于液滴水相运动，使融合液滴内产生混沌流，加速混合；Make the micro-electromagnetic rod reciprocate or circle along the surface of the fusion droplet to drive the magnetic beads in the fusion droplet to move relative to the water phase of the droplet, so that a chaotic flow is generated in the fusion droplet and the mixing is accelerated;
关闭微电磁棒电源，消除磁力作用，使融合液滴与微电磁棒脱离，完成融合液滴混合。Turn off the power of the micro-electromagnetic rod, eliminate the magnetic force, and separate the fusion droplet from the micro-electromagnetic rod to complete the mixing of the fusion droplet.
进一步地，所述含磁珠液滴的固相磁珠与液相的分离操作步骤如下：Further, the separation operation steps of the solid-phase magnetic beads containing magnetic bead droplets and the liquid phase are as follows:
将微电磁棒尖端接触位于超疏水基片表面的含磁珠液滴；The tip of the micro-electromagnetic rod is contacted with the droplet containing magnetic beads on the surface of the superhydrophobic substrate;
开通微电磁棒电源，使其尖端产生磁场，通过磁力作用吸住含磁珠液滴中的磁珠；Turn on the power supply of the micro-electromagnetic rod, make its tip generate a magnetic field, and absorb the magnetic beads in the droplet containing magnetic beads through magnetic force;
上移微电磁棒，使微电磁棒利用吸住的磁珠通过表面张力提起含磁珠液滴；Move the micro-electromagnetic rod up, so that the micro-electromagnetic rod can use the magnetic beads to lift the droplet containing magnetic beads through surface tension;
平移携带含磁珠液滴的微电磁棒至超疏水基片亲水局域位置上方，下移微电磁棒，使含磁珠液滴接触超疏水基片表面亲水局域位置；Translating the micro-electromagnetic rod carrying the droplet containing the magnetic bead to above the hydrophilic local position of the superhydrophobic substrate, and moving the micro-electromagnetic rod downward so that the droplet containing the magnetic bead contacts the hydrophilic local position on the surface of the super-hydrophobic substrate;
再次上移微电磁棒，使微电磁棒吸住的固相磁珠克服液体表面张力作用，从含磁珠液滴中分离出来，完成固相磁珠与液相的分离。Move the micro-electromagnetic rod up again, so that the solid-phase magnetic beads absorbed by the micro-magnetic rod overcome the surface tension of the liquid and separate from the droplet containing magnetic beads, completing the separation of the solid-phase magnetic beads and the liquid phase.
本发明的有益效果为：The beneficial effects of the present invention are:
本发明提供的一种利用微电磁棒操控液滴的方法，无需复杂昂贵的微加工制作过程，也避免了现有永磁体操控液滴方法存在的操控灵活性差、精密度低、实时光学观测困难等缺陷，可准确、灵活、低成本地实现微液滴的多步操控，有望促进数字式液滴微流控技术的发展和应用。The present invention provides a method for manipulating liquid droplets using micro-electromagnetic rods, which does not require complex and expensive micro-machining processes, and also avoids poor control flexibility, low precision, and difficulties in real-time optical observation that exist in the existing methods of manipulating liquid droplets with permanent magnets And other defects, the multi-step manipulation of micro-droplets can be realized accurately, flexibly and at low cost, which is expected to promote the development and application of digital droplet microfluidics technology.
附图说明Description of drawings
为了使本发明的目的、技术方案和有益效果更加清楚，本发明提供如下附图进行说明：In order to make the purpose, technical scheme and beneficial effect of the present invention clearer, the present invention provides the following drawings for illustration:
图1为本发明方法用于输运含磁珠液滴的操作流程示意图。Fig. 1 is a schematic diagram of the operation flow of the method of the present invention for transporting droplets containing magnetic beads.
图2为本发明方法用于融合含磁珠液滴与另一液滴的操作流程示意图。Fig. 2 is a schematic diagram of the operation flow of the method of the present invention for fusing a droplet containing magnetic beads with another droplet.
图3为本发明方法用于促进融合液滴混合的操作流程示意图。Fig. 3 is a schematic diagram of the operation flow for promoting the mixing of fused droplets in the method of the present invention.
图4为本发明方法用于分离含磁珠液滴中固相磁珠与液相的操作流程示意图。Fig. 4 is a schematic diagram of the operation flow for separating the solid-phase magnetic beads and the liquid phase in the droplet containing magnetic beads by the method of the present invention.
具体实施方式detailed description
下面将结合附图，对本发明的优选实施例进行详细的描述。The preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings.
本申请提供的一种利用微电磁棒操控液滴的方法，可以基于磁力、表面张力和重力的竞争，以经过超疏水表面处理的微电磁棒和基片为平台，通过开关微电磁棒实现对含磁珠液滴在超疏水基片表面的吸、放操作，同时结合电磁棒的位移或者液滴座落平台的位移，完成对含磁珠液滴的输运、融合、混合操作以及完成含磁珠液滴中固相磁珠与液相的分离操控。The application provides a method for manipulating liquid droplets by using a micro-electromagnetic rod, which can be based on the competition of magnetism, surface tension and gravity, using the micro-electromagnetic rod and substrate treated with a super-hydrophobic surface as a platform, and realizing the control by switching the micro-electromagnetic rod. The suction and release operation of the droplet containing magnetic beads on the surface of the superhydrophobic substrate, combined with the displacement of the electromagnetic rod or the displacement of the droplet seating platform, completes the transportation, fusion, and mixing operations of the droplet containing magnetic beads Separation and manipulation of solid-phase magnetic beads and liquid phase in magnetic bead droplets.
在本实施方式中，所用微电磁棒由磁芯微棒缠绕通电线圈制成；所述磁芯微棒的截面为圆形，所述磁芯微棒与含磁珠液滴接触的一端为锥形端，且所述锥形端预先经超疏水表面处理。In this embodiment, the micro-rod used is made of a magnetic core microrod wound with an energized coil; shaped end, and the tapered end is pre-treated with a super-hydrophobic surface.
在本实施方式中，所用磁芯的材料为铁、锰-锌铁氧体或镍-锌铁氧体，所述磁芯微棒的直径范围为1～10mm。In this embodiment, the material of the magnetic core is iron, manganese-zinc ferrite or nickel-zinc ferrite, and the diameter of the microrod of the magnetic core is 1-10 mm.
在本实施方式中，所述磁珠为顺磁性微球，直径范围为1～100μm。In this embodiment, the magnetic beads are paramagnetic microspheres with a diameter ranging from 1 to 100 μm.
在本实施方式中，所述超疏水基片的表面包含凹形局域结构或者亲水局域结构。In this embodiment, the surface of the superhydrophobic substrate includes a concave localized structure or a hydrophilic localized structure.
请参阅图1，在本实施方式中，所述含磁珠液滴的输运操作步骤如下：Please refer to Figure 1, in this embodiment, the transport operation steps of the droplet containing magnetic beads are as follows:
将所述微电磁棒的尖端接触位于所述超疏水基片表面的含磁珠液滴；The tip of the micro-electromagnetic rod is contacted with the droplet containing magnetic beads on the surface of the superhydrophobic substrate;
开通所述微电磁棒的电源，使其尖端产生磁场，通过磁力作用吸住所述含磁珠液滴中的磁珠；Turn on the power supply of the micro-electromagnetic rod, make its tip generate a magnetic field, and absorb the magnetic beads in the droplet containing magnetic beads through magnetic force;
上移所述微电磁棒，使所述微电磁棒利用吸住的磁珠通过表面张力提起所述含磁珠液滴；Moving up the micro-electromagnetic rod, so that the micro-electromagnetic rod utilizes the magnetic beads absorbed to lift the droplet containing magnetic beads through surface tension;
平移携带所述含磁珠液滴的微电磁棒至目标位置上方，下移所述微电磁棒，使所述含磁珠液滴接触目标位置基片表面；Translating the micro-electromagnetic rod carrying the magnetic bead-containing droplet above the target position, and moving down the micro-electromagnetic rod so that the magnetic bead-containing droplet contacts the surface of the substrate at the target position;
关闭所述微电磁棒的电源，消除磁力作用，使所述含磁珠液滴与所述微电磁棒脱离，在重力作用下坐落于目标位置，完成液滴输运。Turning off the power supply of the micro-electromagnetic rod, eliminating the magnetic force, so that the droplet containing magnetic beads is separated from the micro-electromagnetic rod, and is located at the target position under the action of gravity to complete the droplet transportation.
请参阅图2，在本实施方式中，所述含磁珠液滴的融合操作步骤如下：Please refer to Figure 2, in this embodiment, the fusion operation steps of the droplet containing magnetic beads are as follows:
将所述微电磁棒的尖端接触位于所述超疏水基片表面的含磁珠液滴；The tip of the micro-electromagnetic rod is contacted with the droplet containing magnetic beads on the surface of the superhydrophobic substrate;
开通所述微电磁棒的电源，使其尖端产生磁场，通过磁力作用吸住所述含磁珠液滴中的磁珠；Turn on the power supply of the micro-electromagnetic rod, make its tip generate a magnetic field, and absorb the magnetic beads in the droplet containing magnetic beads through magnetic force;
上移所述微电磁棒，使所述微电磁棒利用吸住的磁珠通过表面张力提起所述含磁珠液滴；Moving up the micro-electromagnetic rod, so that the micro-electromagnetic rod utilizes the magnetic beads absorbed to lift the droplet containing magnetic beads through surface tension;
平移携带所述含磁珠液滴的微电磁棒至目标液滴上方，下移微电磁棒，使所述含磁珠液滴与目标液滴接触，通过表面张力作用实现融合；Translating the micro-electromagnetic rod carrying the magnetic bead-containing droplet above the target droplet, moving the micro-electromagnetic rod downward to make the magnetic-bead-containing droplet contact the target droplet, and achieve fusion through surface tension;
关闭所述微电磁棒的电源，消除磁力作用，使融合的液滴与微电磁棒脱离，完成液滴融合。Turn off the power supply of the micro-electromagnetic rod, eliminate the magnetic force, and separate the fused droplets from the micro-electromagnetic rod to complete the droplet fusion.
请参阅图3，在本实施方式中，所述含磁珠液滴的混合操作步骤如下：Please refer to Fig. 3, in this embodiment, the mixing operation steps of the droplet containing magnetic beads are as follows:
将微电磁棒尖端接触包含磁珠的融合液滴；Touch the tip of the microelectromagnetic rod to the fused droplet containing the magnetic beads;
开通微电磁棒电源，使其尖端产生磁场，通过磁力作用吸住所述融合液滴中的磁珠；Turn on the power supply of the micro-electromagnetic rod, make its tip generate a magnetic field, and absorb the magnetic beads in the fusion droplet through magnetic force;
使微电磁棒沿所述融合液滴表面往复运动或绕行，带动所述融合液滴中的磁珠相对于液滴水相运动，使融合液滴内产生混沌流，加速混合；Make the micro-electromagnetic rod reciprocate or circle along the surface of the fusion droplet to drive the magnetic beads in the fusion droplet to move relative to the water phase of the droplet, so that a chaotic flow is generated in the fusion droplet and the mixing is accelerated;
关闭微电磁棒电源，消除磁力作用，使融合液滴与微电磁棒脱离，完成融合液滴混合。Turn off the power of the micro-electromagnetic rod, eliminate the magnetic force, and separate the fusion droplet from the micro-electromagnetic rod to complete the mixing of the fusion droplet.
请参阅图4，在本实施方式中，所述含磁珠液滴的固相磁珠与液相的分离操作步骤如下：Please refer to Fig. 4, in this embodiment, the separation operation steps of the solid-phase magnetic beads containing the magnetic bead droplets and the liquid phase are as follows:
将微电磁棒尖端接触位于超疏水基片表面的含磁珠液滴；The tip of the micro-electromagnetic rod is contacted with the droplet containing magnetic beads on the surface of the superhydrophobic substrate;
开通微电磁棒电源，使其尖端产生磁场，通过磁力作用吸住含磁珠液滴中的磁珠；Turn on the power supply of the micro-electromagnetic rod, make its tip generate a magnetic field, and absorb the magnetic beads in the droplet containing magnetic beads through magnetic force;
上移微电磁棒，使微电磁棒利用吸住的磁珠通过表面张力提起含磁珠液滴；Move the micro-electromagnetic rod up, so that the micro-electromagnetic rod can use the magnetic beads to lift the droplet containing magnetic beads through surface tension;
平移携带含磁珠液滴的微电磁棒至超疏水基片亲水局域位置上方，下移微电磁棒，使含磁珠液滴接触超疏水基片表面亲水局域位置；Translating the micro-electromagnetic rod carrying the droplet containing the magnetic bead to above the hydrophilic local position of the superhydrophobic substrate, and moving the micro-electromagnetic rod downward so that the droplet containing the magnetic bead contacts the hydrophilic local position on the surface of the super-hydrophobic substrate;
再次上移微电磁棒，使微电磁棒吸住的固相磁珠克服液体表面张力作用，从含磁珠液滴中分离出来，完成固相磁珠与液相的分离。Move the micro-electromagnetic rod up again, so that the solid-phase magnetic beads absorbed by the micro-magnetic rod overcome the surface tension of the liquid and separate from the droplet containing magnetic beads, completing the separation of the solid-phase magnetic beads and the liquid phase.
具体地，利用本发明提供的利用微电磁棒操控液滴的方法实现微量样品的酶联免疫检测，具体步骤是：Specifically, using the method of using the micro-electromagnetic rod to manipulate the liquid drop provided by the present invention to realize the enzyme-linked immunoassay detection of micro-sample, the specific steps are:
(1)将微电磁棒尖端靠近并接触座落于超疏水基片亲水微区的含磁珠液滴，磁珠表面已预先包被一抗(即捕获抗体)，接通微电磁棒，使液滴中磁珠在微电磁棒的磁场作用下聚集成团，并吸附在微电磁棒尖端。(1) Bring the tip of the micro-electromagnetic rod close to and contact the droplet containing magnetic beads located in the hydrophilic micro-region of the super-hydrophobic substrate. The magnetic beads in the droplet are aggregated into clusters under the action of the magnetic field of the micro-electromagnetic rod, and are adsorbed on the tip of the micro-electromagnetic rod.
(2)上移微电磁棒，使微电磁棒吸住的固相磁珠团克服表面张力作用，从液滴中分离出来。(2) Move the micro-electromagnetic rod up, so that the solid-phase magnetic beads absorbed by the micro-electromagnetic rod overcome the effect of surface tension and separate from the droplet.
(3)平移携带磁珠团的微电磁棒至座落于超疏水基片另一亲水微区的样品液滴上方，下移微电磁棒，使磁珠团与样品液滴接触，通过表面张力作用，磁珠团与样品液融合。(3) Translate the micro-electromagnetic rod carrying the magnetic beads to the top of the sample droplet located on another hydrophilic micro-region of the superhydrophobic substrate, and move the micro-electromagnetic rod down to make the magnetic beads contact with the sample droplet and pass through the surface. Under tension, the magnetic beads fuse with the sample solution.
(4)关闭微电磁棒，使微电磁棒尖端在液滴中往复搅动，分散团聚的磁珠，使磁珠表面捕获抗体与样品中目标分子充分接触，并将液滴放置于37℃温育10～30min。(4) Turn off the micro-electromagnetic rod, make the tip of the micro-electromagnetic rod reciprocate in the droplet to disperse the agglomerated magnetic beads, make the capture antibody on the surface of the magnetic bead fully contact with the target molecule in the sample, and place the droplet for incubation at 37°C 10-30min.
(5)重复类似步骤(1)～(4)的操作流程，将反应后的磁珠从液滴水相中分离，并与座落于超疏水基片另一亲水微区的含二抗(即酶标抗体)液滴融合反应，反应条件为37℃温育10～30min。(5) Repeat the operation process similar to steps (1) to (4), separate the reacted magnetic beads from the droplet water phase, and combine them with the secondary antibody ( (i.e., enzyme-labeled antibody) droplet fusion reaction, the reaction condition is to incubate at 37°C for 10-30min.
(6)重复类似步骤(1)～(4)的操作流程三次，执行三次磁珠清洗，即将反应后的磁珠从液滴水相中分离，并分别与座落于超疏水基片亲水微区的三个洗涤缓冲液液滴融合，搅动使磁珠重悬，每次搅动重悬过程持续30s～1min。(6) Repeat the operation process similar to steps (1) to (4) three times, perform magnetic bead washing three times, that is, separate the reacted magnetic beads from the droplet water phase, and separate them with the hydrophilic microbeads located on the superhydrophobic substrate. The three washing buffer droplets in the zone are fused, agitated to resuspend the magnetic beads, and each agitation and resuspension process lasts for 30s to 1min.
(7)重复类似步骤(1)～(4)的操作流程，将清洗后的磁珠从液滴水相中分离，并与座落于超疏水基片另一亲水微区的含底物液滴融合，执行显色反应，反应条件为37℃温育5～20min。(7) Repeat the operation process similar to steps (1) to (4), separate the washed magnetic beads from the droplet water phase, and mix them with the substrate-containing solution located in another hydrophilic micro-region of the superhydrophobic substrate. The droplets are fused, and the color reaction is performed, and the reaction condition is to incubate at 37°C for 5-20min.
(8)通过移液枪在上述显色反应液滴中加入终止反应液，并重复类似步骤(1)～(2)的操作流程，将磁珠从液滴水相中分离，最后将基片上液滴置于酶标仪中进行信号检测。(8) Add the termination reaction solution into the above-mentioned chromogenic reaction droplet through a pipette gun, and repeat the operation process similar to steps (1) to (2) to separate the magnetic beads from the droplet water phase, and finally put the liquid on the substrate Place the drop in a microplate reader for signal detection.
此外，还可以利用本发明提供的微电磁棒操控液滴的方法实现微量核酸提取，具体步骤是：In addition, the method of manipulating liquid droplets with a micro-electromagnetic rod provided by the present invention can also be used to extract trace amounts of nucleic acid, and the specific steps are:
(1)将微电磁棒尖端靠近并接触座落于超疏水基片亲水微区的含硅基磁珠液滴，接通微电磁棒，使液滴中硅基磁珠在微电磁棒的磁场作用下聚集成团，并吸附在微电磁棒尖端。(1) Bring the tip of the micro-electromagnetic rod close to and contact the droplet containing silicon-based magnetic beads located in the hydrophilic micro-region of the superhydrophobic substrate, and connect the micro-electromagnetic rod so that the silicon-based magnetic beads in the droplet are placed on the surface of the micro-electromagnetic rod. Under the action of a magnetic field, it gathers into clusters and is adsorbed on the tip of the micro-electromagnetic rod.
(2)上移微电磁棒，使微电磁棒吸住的固相硅基磁珠团克服表面张力作用，从液滴中分离出来。(2) Move the micro-electromagnetic rod up, so that the solid-phase silicon-based magnetic beads absorbed by the micro-electromagnetic rod overcome the effect of surface tension and separate from the droplet.
(3)平移携带硅基磁珠团的微电磁棒至座落于超疏水基片另一亲水微区的细胞裂解液液滴上方，下移微电磁棒，使磁珠团与液滴接触，通过表面张力作用，磁珠团与液滴融合。(3) Translate the micro-electromagnetic rod carrying the silicon-based magnetic beads to the top of the cell lysate droplet located on another hydrophilic micro-region of the super-hydrophobic substrate, and move the micro-electromagnetic rod down to make the magnetic beads contact with the droplet , the magnetic beads are fused with the droplets through the action of surface tension.
(4)关闭微电磁棒，使微电磁棒尖端在液滴中往复搅动，分散团聚的硅基磁珠，使磁珠硅基表面与样品中核酸分子充分接触，吸附核酸分子。(4) Turn off the micro-electromagnetic rod, make the tip of the micro-electromagnetic rod reciprocate and stir in the droplet, disperse the agglomerated silicon-based magnetic beads, make the silicon-based surface of the magnetic beads fully contact with the nucleic acid molecules in the sample, and adsorb the nucleic acid molecules.
(5)重复类似步骤(1)～(4)的操作流程三次，执行三次磁珠清洗，即将吸附核酸分子的磁珠从液滴水相中分离，并分别与座落于超疏水基片三个亲水微区的洗涤缓冲液液滴融合，搅动使磁珠重悬，每次搅动重悬过程持续30s～1min。(5) Repeat the operation process similar to steps (1) to (4) three times, perform magnetic bead washing three times, that is, separate the magnetic beads that adsorb nucleic acid molecules from the droplet water phase, and separate them with three beads that are located on the superhydrophobic substrate. The droplets of washing buffer in the hydrophilic micro-area are fused, and stirred to resuspend the magnetic beads, and the resuspension process lasts for 30s to 1min each time.
(6)重复类似步骤(1)～(4)的操作流程，将清洗后的磁珠从液滴水相中分离，并与座落于超疏水基片另一亲水微区的洗脱缓冲液液滴融合，执行洗脱反应，将吸附在磁珠表面的核酸分子洗脱至液滴水相中。(6) Repeat the operation process similar to steps (1) to (4), separate the washed magnetic beads from the droplet water phase, and mix them with the elution buffer located in another hydrophilic micro-region of the superhydrophobic substrate. The droplets are fused, the elution reaction is performed, and the nucleic acid molecules adsorbed on the surface of the magnetic beads are eluted into the aqueous phase of the droplets.
(7)重复类似步骤(1)～(2)的操作流程，将执行洗脱后的磁珠从液滴水相中分离出来，完成核酸提取过程。(7) Repeat the operation process similar to steps (1)-(2), separate the eluted magnetic beads from the droplet water phase, and complete the nucleic acid extraction process.
由上可见，本发明的有益效果为：As can be seen from the above, the beneficial effects of the present invention are:
本发明提供的一种利用微电磁棒操控液滴的方法，无需复杂昂贵的微加工制作过程，也避免了现有永磁体操控液滴方法存在的操控灵活性差、精密度低、实时光学观测困难等缺陷，可准确、灵活、低成本地实现微液滴的多步操控，有望促进数字式液滴微流控技术的发展和应用。The present invention provides a method for manipulating liquid droplets using micro-electromagnetic rods, which does not require complex and expensive micro-machining processes, and also avoids poor control flexibility, low precision, and difficulties in real-time optical observation that exist in the existing methods of manipulating liquid droplets with permanent magnets And other defects, the multi-step manipulation of micro-droplets can be realized accurately, flexibly and at low cost, which is expected to promote the development and application of digital droplet microfluidics technology.
最后说明的是，以上优选实施例仅用以说明本发明的技术方案而非限制，尽管通过上述优选实施例已经对本发明进行了详细的描述，但本领域技术人员应当理解，可以在形式上和细节上对其作出各种各样的改变，而不偏离本发明权利要求书所限定的范围。Finally, it should be noted that the above preferred embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail through the above preferred embodiments, those skilled in the art should understand that it can be described in terms of form and Various changes may be made in the details without departing from the scope of the invention defined by the claims.
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| CN113908897A (en) * | 2021-11-16 | 2022-01-11 | 中山大学 | A microfluidic device for magnetic excitation to realize droplet manipulation and manipulation method thereof |
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| CN115722284A (en) * | 2022-11-21 | 2023-03-03 | 西南科技大学 | Structure for oriented transportation and large-area collection of micro-droplets and preparation method |
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