一种油水分离的微流控芯片及其制备方法技术领域 本发明涉及一种油水分离的微流控芯片及其制备方法，该微流控芯片表面有微结构和微通道，在微通道的左右两侧分别进行亲水和亲油修饰，在电场的作用下，油水混合溶液发生破乳，水往亲水侧移动，油往亲油侧移动，最后实现油水分离，主要应用于变压器油、 发电厂透平油、海上原油泄漏事故处理、油田落地油、海上钻井平台浮油、钢铁厂平流油、炼油厂大量含油废水等相关领域。 One kind of oil-water separation microfluidic chip and TECHNICAL FIELD  The present invention relates to a micro-fluidic chip and a method for preparing oil-water separation, the microfluidic chip surface microstructures and microchannels in the micro- left and right channels are respectively hydrophilic and lipophilic modified, under the action of an electric field, the occurrence of breaking oil-water mixed solution, hydration of the hydrophilic side to move, oil moves towards lipophilic side, finally to achieve water separation, mainly used in transformer oil , power plant turbine oil, offshore oil spill treatment, ground oil fields, the oil slick offshore drilling platforms, steel mills stratospheric oil refinery a lot of oily wastewater and other related fields. 背景技术 20世纪80年代后期，为了建立更类似于体内环境的培养体系，尽可能使体外环境与体内环境相吻合，从而使细胞间能相互沟通信息，相互支撑生长增殖，人们在细胞培养技术的基础上发展出了细胞共培养技术。  20 in the late 1980s, in order to establish a training system more similar to the internal environment, as far as possible the in vitro environment and internal environment is consistent, so that cells can communicate information between each other, mutual support growth and proliferation of cells in people the development of basic technical training on the cell co-culture techniques. 细胞共培养技术是将2种或2种以上的细胞共同培养于同一环境中，由于其具有更好地反映体内环境的优点，所以这种方法被广泛应用于现代细胞研究中。 Cell co-culture techniques is one or two or more cells co-cultured in the same environment, since it has the advantage of better reflect the in vivo environment, so this method is widely used in modern cell research. 目前，细胞共培养技术最多应用于骨细胞和神经细胞。  At present, most cell co-culture technology in bone and nerve cells. 细胞共培养体系主要通过两种方法建立：①直接共培养体系，即将2种或2种以上的细胞同时或分别接种于同一孔中，不同种类的细胞之间直接接触；②间接共培养体系，即将2种或2种以上的细胞分别接种于不同的载体上，然后将这两种载体置于同一培养环境之中，使不同种类的细胞共用同一种培养体系而不直接接触。 Coculture established mainly two methods: ① direct co-culture system, is about two or more kinds of cells were inoculated simultaneously or in the same well, the direct contact between the different types of cells; ② indirect co-culture system, is imminent or more kinds of two kinds of cells were seeded on different carriers, then these two vectors placed in the same culture environment in which different types of cells sharing the same without directly contacting the culture system.  共培养体系主要作用：诱导细胞向另一种细胞分化；诱导细胞自身分化；维持细胞功能和活力；调控细胞增殖；促进早期胚胎发育和提高代谢产物产量。  The primary role of co-culture system: induce cell differentiation to another cell; itself induce cell differentiation; regulation of cell proliferation;; maintain cell function and viability promote early embryonic development and increase the metabolic product yield. 从上世纪30年代开始，细胞培养逐渐成为研究人员实验过程中不可缺少的重要步骤，其载体工具：培养皿/ 培养瓶也逐步被大家所认可，成为了一种常规的实验耗材。 From the beginning of the 1930s, cell culture experiment researchers gradually become an important and indispensable step in its support tools: dish / flasks are also gradually being recognized by everyone, it has become a routine laboratory supplies. 虽然到目前为止，很多科学家认为培养皿/培养瓶的这种体外培养条件与体内生长环境有着显著的不同,但是由于没有更好的培养载体来改变这个现状，所以生物学家们也只能退而求次的默认这种情况的存在。 Although so far, many scientists believe that dish / flasks of this culture conditions in vitro and in vivo growth environment has a significant difference, but the lack of a better culture carrier to change the status quo, so biologists can only retreat and there is a default in this case seek times. 但是近来，一种Petaka细胞培养系统的问世，获得美国、欧洲广大生物科学家的青睐，培养皿/培养瓶的一些致命缺陷又再一次被提及，成为研究人员热烈讨论的话题之一。 But recently, the advent of one kind Petaka cell culture system, the US, Europe favor the majority of biological scientists, dish / flasks, some fatal flaw once again been mentioned as one of the researchers topic of lively discussion.  常规的细胞培养皿很难完成多种细胞的共培养，因此，发展一种便捷、快速、高效、 低成本的多细胞培养技术，是细胞生物学等领域的迫切需求。  Conventional cell culture dish more difficult to achieve co-cultured cells, and therefore, the development of a convenient, fast, efficient, low-cost multi-cell culture technology is an urgent demand in the field of cell biology. 近年来，微流控芯片分析技术已成为分析化学中一个重要的研究方向，是其中最活跃的一支，无论是在科研还是应用领域都获得了广泛的重视。 In recent years, microfluidic analysis technology has become an important analytical chemistry research, it is one of the most active one, both in research or applications have gained widespread attention. 微流控芯片作为一种新型的分析检测平台，具有高通量、集成化、 多重平行分析、便携式、易操作、成本低等优点，已经在众多领域获得了广泛应用。 Microfluidic chip as a new platform for analysis and detection, high throughput, integrated, multiple parallel analysis, portable, easy to operate, low cost, has been widely used in many fields. 然而，采用微流控芯片，在其表面制备微结构和微通道，依靠微通道中多层液体之间的层流效应驱动样品微流体，同时完成多种细胞的植入技术，目前在多种细胞共培养的应用领域尚未有实质性的突破。 However, with the microfluidic chip, in its micro-structure and micro channel surface preparation, depend on flow effect between the layers of the multilayered liquid microchannels of microfluidic sample drive, while the completion of more cells implantation techniques, various currently cell co-culture of the applications have not been substantive breakthrough.  多种细胞共培养微流控芯片能将2种或2种以上的细胞共同培养于同一环境中， 能更真实地反映人体组织细胞之间的互相影响，有利于实验者观察细胞与细胞之间互相作用，特别是多个细胞对一个细胞的影响，也有利于快速筛选新药的疗效和毒性。  more coculture microfluidic chip capable of two or more kinds of cells co-cultured in the same environment, more truly reflect the mutual influence between human tissue cells, and the cells were observed facilitate experimenter interaction, particularly those affecting cell between multiple cells to a cell, but also conducive to the rapid screening of drug efficacy and toxicity. 发明内容 本发明的目的是提供了一种油水分离的微流控芯片及其制备方法，该微流控芯片表面有微结构和微通道，在微通道的左右两侧分别进行亲水和亲油修饰，在电场的作用下， 油水混合溶液发生破乳,水往亲水侧移动,油往亲油侧移动,最后实现油水分离。 SUMMARY OF THE INVENTION  The object of the present invention is to provide a microfluidic chip and method for preparing oil-water separation, the microfluidic chip surface microstructures and microchannels, hydrophilic left and right sides, respectively, of the microchannel and lipophilic modified, under the action of an electric field, the occurrence of breaking oil-water mixed solution, the water moves to the side of a hydrophilic, lipophilic side toward the oil transfer, to achieve the final oil-water separation. 微流控芯片由刻有微米级别的微结构和微通道的芯片和粘性薄膜封合而成，微结构和微通道通过微加工技术制备。 Microfluidic chip by the engraved micron microstructure and micro-channel chips and the adhesive membrane sealed together, the microstructure and the microchannel prepared by micro-machining techniques.  为实现上述目的，本发明采用以下的操作步骤： (I)用计算机辅助设计软件设计和绘制微流控芯片中各层芯片的微结构和微通道图形。  To achieve the above object, the present invention employs the following steps:  (I) and the design drawing microfluidic microstructure and micro-channel chip graphics controller chip in the layers of computer-aided design software.  (2)通过微加工技术在各层微流控芯片基材表面和粘性薄膜上加工所需的微结构和微通道，包括进样孔、分离主通道和分离分通道。  (2) by micromachining techniques required for processing layers on a surface of the substrate and the microfluidic chip adhesive membrane microstructures and microchannels, including the injection hole, and the separation of the main channel separated sub-channel.  (3)利用双层粘性薄膜，将各层离心式微流控芯片对齐、粘合、加压封合，组成油水分离的微流控芯片。  (3) by the double adhesive film, the microfluidic chip alignment layers centrifugation, adhesive, pressure sealing, consisting of oil-water separation microfluidic chip.  (4)将油水混合溶液从样品池加入，施加一定电场，使油水混合溶液进行破乳。  (4) from the oil-water mixture was added to the sample cell, applying a constant electric field, so that demulsification of oil and water mixed solution.  (5)溶液迅速破乳后，利用亲水和亲油界面的同性相吸原理，完成油水的分离。  (5) After the solution was rapidly breaking, with a hydrophilic and lipophilic interface like attracts principle, complete separation of oil and water.  本发明中，油水分离的微流控芯片的芯片基材可以是PMMA、PC、PVC、C0C、铜、铝、不锈钢、硅片、玻璃圆片，也可是市售的各类普通CD光盘。  In the present invention, the oil-water separation microfluidic chip substrate chip may be PMMA, PC, PVC, C0C, copper, aluminum, stainless steel, silicon wafer, glass wafer, but also various types of commercially available ordinary CD CD.  本发明中，油水分离的微流控芯片和粘性薄膜的微结构和微通道可以通过数控铣刻、激光刻蚀、LIGA技术、模塑法、热压法、化学腐蚀制备，也可用软刻蚀技术制备。  In the present invention, the oil-water separation microstructures and microchannels and the microfluidic chip adhesive film may be cut by CNC milling, laser etching, the LIGA technology, molding, pressing, preparing chemical etching, can also be used preparation of soft lithography.  本发明中，油水分离的微流控芯片是由两层芯片组成，各层芯片之间用粘性薄膜贴合，粘性薄膜可以是双层力致粘性薄膜，也可是普通双面胶薄膜。  In the present invention, the oil-water separation microfluidic chips is composed of two layers, between the layers of adhesive film bonded chips, the adhesive film may be a double adhesive film actuator force, but also the ordinary double-sided adhesive film .  本发明中，油水分离的微流控芯片上的样品溶液的驱动依靠油水溶液储液罐与进样孔之间的液差所产生的重力。  In the present invention, the oil-water separation of the sample solution on the driven microfluidic chip relies on a difference between the fluid reservoir and the inlet of oil wells an aqueous solution produced by gravity.  本发明中，油水分离的微流控芯片的油水破乳是依靠微通道两侧之间形成的电场来完成的。  In the present invention, the water demulsifying oil-water separation microfluidic chip is to rely on an electric field formed between both sides of the microchannels to complete.  本发明中，油水分离的微流控芯片的微通道两侧的表面分别进行了亲水和亲油的表面修饰。  In the present invention, the surface of the water on both sides of the microchannel of the microfluidic chip separation were surface-modified hydrophilic and lipophilic.  本发明中，油水分离的微流控芯片进行微通道表面修饰的亲水剂是 本发明中，油水分离的微流控芯片进行微通道表面修饰的亲油剂是 本发明提出的油水分离的微流控芯片及其制备方法，操作简单、实现了油水混合溶液的快速破乳和迅速分离，降低了试剂与样品的用量，简化了分离过程，具有便携、经济、 快速、高效的特点，在油水分离的相关领域中具有良好的应用前景。  In the present invention, the oil-water separation microfluidic microchannel chip surface-modified hydrophilic agent is  the present invention, the oil-water separation microfluidic chip surface of the microchannel is modified pro-oil [0022 ] the microfluidic chip of the oil-water separation and preparation method proposed by the present invention, simple operation, to achieve a rapid breaking of oil-water mixture solution and quickly separated, to reduce the amount of reagent and sample to simplify the separation process, portability, economical fast, efficient features, has a good prospect in the relevant field of oil-water separation. 附图说明 图1.油水分离的微流控芯片的结构示意图。 BRIEF DESCRIPTION OF schematic configuration  FIG. 1. The water separator microfluidic chip.  a.亲水界面，b.亲油界面，c.电场正极，d.电场负极，e.电源。  a. A hydrophilic interface, b. Lipophilic interface, c. A positive electric field, d. A negative electric field, e. Power. 具体实施方案 实施例1 用计算机辅助设计软件设计和绘制离心式微流控芯片的两层芯片的微结构和微通道图形。 Specific embodiments  Example 1  Design and draw graphics microstructure and centrifuged two microchannel chip microfluidic chip using computer aided design software. 利用数控CNC系统加工制备两层圆片状聚甲基丙烯酸甲酯（PMMA)芯片的微结构和微通道，分别用自来水、蒸馏水清洗各层芯片，并用乙醇擦拭芯片表面残留的指纹、油溃等污溃。 NC CNC machining system prepared using two disc-shaped polymethyl methacrylate (PMMA) and microstructure chip microchannels, respectively, tap water, distilled water layers chip, and the chip with ethanol wipe fingerprints remaining on the surface, like oil collapse sewage collapse. 在双面胶薄膜上，用刻字机加工制备所需的微结构和微通道。 On the double-sided adhesive film, with a cutting plotter process for preparing the desired microstructure and microchannels. 将两层芯片小心对齐、粘合、加压封合，制成油水分离的微流控芯片。 The two layers were carefully aligned with the chip, adhesive, pressure sealing, made of oil-water separation microfluidic chip. 将油水溶液储液罐与进样注入孔相连， 利用液差加入待分离的油水混合溶液，施加电场，油水混合溶液在分离主通道迅速破乳，水组分向亲水界面移动，油组分向亲油界面移动，最后实现油水分离。 The oil and aqueous sample reservoir is connected to the injection hole, using a mixed solution of water were added to the difference to be separated, an electric field is applied, the mixed solution was separated oil demulsifying main channel quickly, the water moves to the hydrophilic interface component, the oil component lipophilic interface moves to finally realize the oil-water separation.  实施例2 用计算机辅助设计软件设计和绘制离心式微流控芯片的两层芯片的微结构和微通道图形。  Example 2  The microstructures and microchannels graphic design and rendering centrifugal microfluidic chip using computer aided design software embodiment of the two chips. 利用数控CNC系统加工制备两层圆片状聚碳酸酯（PC)芯片的微结构和微通道， 分别用自来水、蒸馏水清洗各层芯片，并用乙醇擦拭芯片表面残留的指纹、油溃等污溃。 NC CNC machining system prepared using two disc-shaped polycarbonate (PC) and microstructure chip microchannels, respectively, tap water, distilled water chip layers, and washed with ethanol fingerprints, dirt, etc. collapse crushed oil remaining on the surface wiped chip. 在双面胶薄膜上，用刻字机加工制备所需的微结构和微通道。 On the double-sided adhesive film, with a cutting plotter process for preparing the desired microstructure and microchannels. 将两层芯片小心对齐、粘合、加压封合，制成油水分离的微流控芯片。 The two layers were carefully aligned with the chip, adhesive, pressure sealing, made of oil-water separation microfluidic chip. 将油水溶液储液罐与进样注入孔相连，利用液差加入待分离的油水混合溶液，施加电场，油水混合溶液在分离主通道迅速破乳，水组分向亲水界面移动，油组分向亲油界面移动，最后实现油水分离。 The oil and aqueous sample reservoir is connected to the injection hole, using a mixed solution of water were added to the difference to be separated, an electric field is applied, the mixed solution was separated oil demulsifying main channel quickly, the water moves to the hydrophilic interface component, the oil component lipophilic interface moves to finally realize the oil-water separation.