CN104492508B - An apparatus and method control an amount of an ultrafine liquid droplets remaining - Google Patents

An apparatus and method control an amount of an ultrafine liquid droplets remaining Download PDF

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CN104492508B
CN104492508B CN 201410640394 CN201410640394A CN104492508B CN 104492508 B CN104492508 B CN 104492508B CN 201410640394 CN201410640394 CN 201410640394 CN 201410640394 A CN201410640394 A CN 201410640394A CN 104492508 B CN104492508 B CN 104492508B
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liquid
taken
droplets
probe
droplet
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CN104492508A (en )
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方群
祝莹
郭晓利
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浙江大学
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Abstract

本发明公开了一种基于液体残留的超微量液滴操控装置,包括:多孔板;微流控液滴阵列芯片;取液探针,该取样探针上具有支持部分、以及位于支持部分底端的取液端;所述取液端表面对被取液体具有亲和性,所述支持部分外表面对被取液体不具备亲和性;切换机构,依次实现取液探针对被取液体的取样操作以及将所取被取液体置于微流控液滴阵列芯片内的放样操作。 The present invention discloses a liquid ultrafine droplet amount based residual control device, comprising: a porous plate; microfluidic droplet array chip; a tapping probe, having a supporting portion of the sample probe, and the bottom end portion of the support located take liquid end; the end surface of the liquid taken is taken to have an affinity for the liquid, the outer face of the support portion does not have affinity for the liquid is taken; switch mechanism sequentially to achieve the probe was taken for sampling the liquid to be taken the operation and be taken for liquid placed within the loft operation microfluidic droplet array chip. 本发明还公开了一种基于液体残留的超微量液滴操控方法。 The present invention also discloses ultrafine droplet control method based on the amount of residual liquid. 本发明具有皮升以下,甚至低至飞升到阿升级的体积精度,有效降低了化学和生物反应和分析中的样品和试剂消耗,节省了实验成本;无需采用注射泵、蠕动泵等驱动部件和毛细管等管路,系统操作简单、可靠性高、易于实现阵列化和自动化。 The present invention has picoliter or less, even as low volume precision soar upgrade A, effectively reducing the chemical and biological reactions and the analysis of sample and reagent consumption, cost saving experiment; driven member without using a syringe pump, peristaltic pump, and capillary conduit, the system is simple, high reliability, easy to implement and automate array.

Description

一种基于液体残留的超微量液滴操控装置及方法 An apparatus and method control an amount of an ultrafine liquid droplets remaining

技术领域 FIELD

[0001]本发明属于分析化学中的微流控液滴分析领域,具体是涉及一种基于液体残留的超微量液滴操控装置及方法。 [0001] The present invention belongs to analysis of microfluidic droplets in analytical chemistry, and particularly relates to apparatus and method for ultrafine liquid remaining amount of liquid droplets based on manipulation.

背景技术 Background technique

[0002]微量的液体操控方法和装置在当前分析化学领域的发展中起着越来越重要的作用。 [0002] method and apparatus for handling liquid trace the development of the field of chemistry plays an increasingly important role in the current analysis. 一方面,液体操控体积的降低减少了对珍稀样品的需求量,显著降低了实验难度和成本。 In one aspect, reducing the volume of control liquid reduces the demand for rare samples significantly reduces the difficulty and cost of the experiment. 另一方面,微体系的体积效应避免了对超微量样品的过度稀释,从而实现了常规大体积体系难以完成的化学分析任务,如单细胞和单分子分析。 On the other hand, the volume effect microarchitecture avoids excessive dilution of the sample minim, thereby realizing a conventional bulk chemical analysis task difficult to complete the system, such as a single cell and single-molecule analysis. 微流控芯片技术通过微机电加工技术,在玻璃或者塑料等基片上加工网络化的微米级通道,并结合压力、电、磁、光、声、热等驱动和控制方法对通道内的超微量液体进行操控,从而完成微量生化反应与分析。 Microfluidic chip techniques MEMS processing technology, on a glass or plastic and other substrate processing network of micron scale channels, combined with pressure, an electronic, magnetic, optical, acoustic, thermal drive and control method ultramicro amount within the channel liquid handling, thus completing the micro-biochemical reactions and analysis. 微流控技术已经成为微量液体操控的平台技术,广泛应用于化学和生物分析、化学合成、药物筛选、医学诊断、组学研究等领域。 Microfluidic technology has become a platform technology of micro-liquid handling, it is widely used in chemical and biological analysis, chemical synthesis, drug screening, medical diagnostics, genomics research and other fields.

[0003]基于液滴的微流控技术是近年来快速发展的一种新型微量液体操控和生化分析技术。 [0003] is a novel handling trace amount of liquid and the rapid development in recent years, biochemical analysis based microfluidics droplets. 液滴微流控技术通过对微通道或微结构中不互溶的两相液体的操控,实现皮升至纳升级油包水型(或水包油型)液滴反应器的定量生成、混合、分裂、筛选等,从而可在超微量体积上完成常规化学和生物反应所需的液体操控操作。 Droplet microfluidics by manipulation of the micro-channel or two-phase liquid immiscible microstructure achieve picoliter to nanoliter quantitative generated water-in-oil type (or oil in water) droplets of the reactor, mixing, splitting, filtering, etc., so that the liquid can complete the desired operation control routine in the chemical and biological reactions minim volume. 与连续流(单相)微流控技术相比,液滴微流控技术消除了微反应器中反应物或产物的稀释与扩散,且显著提高了反应器中传质和传热速度。 Compared with the continuous flow (single phase) microfluidics, microfluidic droplets dilution technique eliminates the diffuser microreactor reactants or products, and significantly improves the rate of heat transfer and mass transfer in the reactor. 同时,生物兼容的油水界面也为超微量生化反应提供了温和均一的微环境,有效提尚了其反应效率。 Meanwhile, bio-compatible oil-water interface is also provided modest uniform microenvironment for ultra trace biochemical reactions, yet effectively improve the efficiency of the reaction.

[0004]目前,多数的液滴系统均采用具有T型或者十字聚焦型通道的微芯片进行液滴的形成和操控。 [0004] Currently, the majority of droplets having a microchip system are T-shaped or cross-shaped channel is focused droplet formation and handling. 一般通过调节通道的尺寸和两相流速来调节液滴的大小和生成频率(ThorsenT,Roberts R W1Arnold FH,Quake S R.,Phys.Rev.Lett.,2001,86:4163〜4166;Anna SL,Bontoux N, Stone H A.,Appl.Phys.Lett.,2003,82:364〜366)。 Typically adjusted by adjusting the size of the two-phase flow velocity and the size of the droplets and channel frequency generation (ThorsenT, Roberts R W1Arnold FH, Quake S R., Phys.Rev.Lett, 2001,86: 4163~4166; Anna SL,. Bontoux N, Stone H A., Appl.Phys.Lett, 2003,82:. 364~366). 为了向液滴内注入样品或者试剂溶液进行生化反应测定,主要采用两类方法:一种是采用T型通道将连续试样注入到不同的液滴中(Zheng B,Ismagilov R F.,Angew.Chem.1nt.Ed.,2005,44:2520〜2523);另一种是采用流体压力或电动的方法将两个或者多个含有不同组分的液滴进行融合(NiuX1Gulati S1Edel JB,deMello A J.,Lab Chip,2008,8:1837〜1841;Mazutis L1AraghiA F1Miller OJ,Baret J C1Frenz L,Janoshazi A,Taly V1Miller BJ,Hutchison JB,Link D,Griffiths AD,Ryckelynck M.,Anal.Chem.,2009,81:4813〜4821)。 For injection of the sample or biochemical reaction measuring reagent solution into the droplets, mainly two methods: one is to use a continuous T-shaped channel sample was injected into different droplet (Zheng B, Ismagilov R F., Angew. Chem.1nt.Ed., 2005,44: 2520~2523); another method is the use of a fluid pressure or electric droplet containing two or more different components are fused (NiuX1Gulati S1Edel JB, deMello a J ., Lab Chip, 2008,8: 1837~1841; Mazutis L1AraghiA F1Miller OJ, Baret J C1Frenz L, Janoshazi A, Taly V1Miller BJ, Hutchison JB, Link D, Griffiths AD, Ryckelynck M., Anal.Chem, 2009,. 81: 4813~4821). 这类基于微通道网络的液滴操控技术具有液滴生成和操控速度快,自动化程度高等优点,特别适合需要进行大规模液滴操控的应用场合。 Such control technology based on microchannel network droplets having droplet formation and manipulation applications speed, high degree of automation, particularly suitable for large-scale droplet manipulation needs. 然而,这类液滴技术也存在一些明显的局限性。 However, such droplets technology there are some obvious limitations. 首先,这类方法难以快速生成不同化学组成和浓度的液滴,限制了其在以多样品为主要特征的高通量药物筛选等方面的应用。 First, such methods it is difficult to quickly generate different chemical composition and concentration of the droplets, which limits its application in high-throughput screening of multiple samples or the like to the main characteristic of. 其次,为了实现试样加入和液滴融合所采用的压力或电动方法,均需要对液滴和油相的组成、液滴的流速、位置进行精确控制,操作难度大,其所需芯片的成本也较高。 Secondly, in order to achieve a pressure or electric fusion method a sample is added and the droplet employed, are required for the composition of the oil phase droplets and the droplet velocity, precise control of the location, a large operation is difficult, the cost required for chip which higher. 此外,由于液滴存储于封闭的芯片通道中,难以直接从液滴中取样进行下一步的分析测定。 Further, since the storage droplets, it is difficult to sample directly from the droplets in a closed chip channel measured the next analysis.

[0005]最近,申请人所在的研究组发展了一种基于平面二维液滴阵列的液滴操控方法(方群,祝莹,张云霞,一种具有皮升级精度的自动化微液滴阵列筛选系统的使用方法,申请号:201210589055;祝莹,方群,张云霞,朱隨娜,一种半接触式的油下液滴连续点样和加液方法,申请号:201410161574;Zhu Y.,Zhang YX,Cai LF,Fang Q.,Anal.Chem.,2013,85:6723〜6731)。 [0005] Recently, the applicant's research group developed a method for manipulating a droplet (a two-dimensional plane based Fangqun droplet array, Zhu Ying Zhang Yunxia, ​​having picoliter droplet array precision automated screening system method of use, application number: 201 210 589 055; Zhu Ying Fang group, Yun-Xia Zhang, Zhu with Na, spotting droplets continuously and the charging method, a semi application No. under contact oil: 201410161574; Zhu Y., Zhang YX, Cai LF, Fang Q., Anal.Chem, 2013,85:. 6723~6731). 区别于在封闭通道中生成和操控液滴的方法,该方法在覆盖有一定厚度油相的平面芯片上进行液滴的操控。 Different from the method for generating and manipulating a droplet in a closed passage, which manipulate covered with droplets on planar chip with a thickness of an oil. 该方法采用基于注射栗、蠕动栗或者其他基于压力驱动的双向液体驱动系统进行液体的定量抽取,在拉尖的毛细管通道中生成液滴,并结合高精度三维平移台将毛细管中的液滴滴加至平面芯片上。 The quantitative method using the liquid injection chestnut extract, a peristaltic or other Li-based two-way pressure-driven fluid drive system based on the generated droplets icicling capillary channel, and combining high precision 3D translation stage liquid drops capillary was added to the chip plane. 由于覆盖有一层油相的液滴储存芯片是一种开放式的液滴系统,试样的加入和取样可直接通过将毛细管插入液滴的方法实现。 Since the cover layer of oil phase droplets storage chip it is an open system drops, the sample was added and the sample can be achieved directly by the method of inserting the capillary the droplet. 因此,基于二维液滴阵列的液滴操控系统解决了常规液滴系统难以生成大量不同化学组成液滴、加液复杂、成本高、难以取样的特点,在高通量化学与生物分析和筛选、单细胞分析、医学诊断等方面具有重大的应用潜力。 Thus, a two-dimensional array of droplets droplet control system to solve the conventional droplet-based system is difficult to generate a large number of droplets of different chemical composition, dosing complex, costly, difficult sampling characteristics, chemical and biological analysis of high-throughput screening and , single-cell analysis, and other aspects of medical diagnosis has important potential applications. 然而,由于采用基于注射栗、蠕动栗等压力驱动技术的液体量取方法,受到机械运动精度和毛细管通道内界面张力的影响,其系统的液体操控体积通常在数十皮升至纳升级,难以精确操控低于皮升的超微量液体体积。 However, since the liquid amount of the injection method of driving Based chestnut, chestnut, etc. peristaltic pressure, by mechanical movement of the capillary and the inner passage Accuracy of interfacial tension, the liquid handling system, which volume is usually in the tens picoliter to nanoliter difficult precise control of the amount of ultrafine below picoliter volumes of liquid. 尤其是针对多样品的可在低于皮升级的水平进行超微量液体操控的系统,目前尚无文献报道。 Especially for ultra-fine system can be manipulated more than the amount of liquid sample in the skin below the level of upgrade, there is no literature.

发明内容 SUMMARY

[0006]本发明的目的是提供一种可具有皮升(10—12升)以下,甚至低至飞升到阿升级(10—15到10—18升)液体操控精度的基于液体残留的超微量液滴操控装置该装置采用表面对被取液体具有亲和作用的实心取液探针,利用其在脱离被取液体时在取液端表面残留少量液体的现象,在微流控液滴系统中实现超微量的液滴生成、取液、混合和转移等复杂化液体操控。 [0006] The object of the present invention is to provide a having a picoliter (10-12 liters) or less, even down to soar A upgrade (10-15 to 10-18 liters) of liquid ultra precision control based on the amount of liquid remaining the device uses droplet control device for solid surface liquid has been taken to take effect liquid affinity probe, with its end surface taking small amount of residual liquid in the liquid when the liquid is taken off phenomenon in droplet microfluidic system ultratrace achieve droplet formation, was taken, and a mixed liquid metastasis complicated manipulation.

[0007]本发明同时提供了一种基于液体残留的超微量液滴操控方法,该方法适用于超微量的化学和生物分析、高通量药物筛选、蛋白质结晶条件筛选、单细胞分析、单分子分析等需要超微量液体操控的场合中。 [0007] The present invention also provides a method of control based on the amount of liquid droplets ultra residual liquid, which is suitable for ultratrace chemical and biological analysis, high throughput drug screening, screening of protein crystallization conditions, single cell analysis, single molecule analysis and other applications requiring ultra-fine control of the amount of liquid.

[0008] 一种基于液体残留的超微量液滴操控装置,包括: [0008] Based on the amount of ultrafine liquid droplets remaining control device, comprising:

[0009]多孔板,该多孔板上设有一个或多个用于盛放被取液体的孔槽; [0009] perforated plate, the porous plate provided with a plurality of holes or slots for accommodating a liquid to be taken;

[0010]微流控液滴阵列芯片,该微流控液滴阵列芯片上设有一个或多个用于承载液滴的微结构; [0010] microfluidic droplet array chips, has a microstructure for carrying one or more droplets on the microfluidic chip array of droplets;

[0011 ]取液探针,该取样探针上具有支持部分、以及位于支持部分底端的取液端;所述取液端表面对被取液体的亲和性大于等于所述支持部分外表面对被取液体的亲和性; [0011] probe was taken, the sample probe having a support portion and positioned to support the bottom end of a tapping portion; end of the extracting solution is taken to the liquid surface of the affinity greater than or equal to the outer face of the support section the affinity for the liquid is taken;

[0012]切换机构,依次实现取液探针对被取液体的取样操作以及将所取被取液体置于微流控液滴阵列芯片内的放样操作。 [0012] switching means for sequentially sampling probe to achieve a tapping operation is taken and the taken liquid is taken the liquid disposed in the loft operation microfluidic droplet array chip. 所述切换机构可采用三维运动工作台。 The switching mechanism may be employed a three-dimensional moving table.

[0013]所述的取液探针为采用玻璃、石英、金属或者高分子聚合物材料加工,所述取液探针的外形为实心针形或实心柱形。 [0013] The probe was taken as glass, quartz, metal, or polymer material processing, taking the shape of the probe is a solid solution form or solid cylindrical pin. 所述取液探针分为取液端和支持部分两部分。 The probe was taken into a tapping end supporting portion and two parts. 所述取液端的表面对被取液体具有亲和性,或者取液端的表面被处理成对所取液体具有亲和性的表面,所述的支持部分的表面对被取液体不具有亲和性。 The end surface take liquid having affinity for liquid is taken, or the end surface of the liquid to be treated to take up in pairs having affinity for liquid surface, the surface of the support portion having no affinity for the liquid to be taken . 或者支持部分的表面被处理成对所取液体不具有亲和性的表面。 Or the surface of the supporting portion of the liquid to be treated, taken in pairs having no affinity surface.

[0014]所述取液探针的横截面的形状为圆形,或椭圆形,或三角形,或四边形,或其他多边形。 Shape [0014] The probe was taken cross section is circular, or oval, or triangular or quadrangular, or other polygonal. 所述取液探针的横截面的面积范围是100平方纳米至1000平方毫米。 Take the cross-section area of ​​the probe was in the range of 100 nm to 1000 mm2 square. 所述取液探针的长度范围是I微米至10厘米;采用多个取液探针构成一维取液探针阵列,或者二维取液探针阵列,可完成多样品操控。 The length of the probe take liquid ranges I micron to 10 cm; using a plurality of probes constituting the extract solution was taken one-dimensional array probe or a two-dimensional array probe was taken, control can be done of multiple samples.

[0015]所述取液端表面的面积为100平方纳米至1000平方毫米。 [0015] The end surface of the liquid takes an area of ​​100 square nanometers to 1000 mm2. 根据所取液体的性质,选择合适的材料加工取液探针,分别选择所述取液探针上合适的区域作为取液探针的取样端和支持部分;如所述取液探针的取液端和支持部分的表面对被取液体的亲和性不能达到要求,则采用物理的研磨或抛光或光照方法,或者采用化学的腐蚀或表面吸附或表面接枝方法,分别对取液探针的取液端和支持部分的表面进行处理,使得取液端的表面对被取液体具有亲和性,支持部分的表面对被取液体不具有亲和性。 Depending on the nature of the liquid taken, select suitable material extracting solution probes, were selected on the appropriate region was taken as the sample probe and the support end portion of the probe was taken; taken as the probe was taken end surface and the liquid affinity support portion of the liquid is taken not meet the requirements, the use of physical methods or light polishing or grinding, or by etching or surface adsorption or chemical surface grafting method, were taken for liquid probe It was taken and the surface for supporting an end portion of the process, so that the surface side of the liquid taken is taken for having an affinity for the liquid, the surface of the support portion does not have affinity for liquid.

[0016]所述微流控液滴阵列芯片为采用玻璃、石英、金属或者高分子聚合物材料加工,所述微流控液滴阵列芯片上加工有承载液滴的微结构。 The [0016] microfluidic droplet array chip of glass, quartz, metal or polymer materials processing, the microfluidic processing structure carries a micro droplet on the droplet array chip. 所述微流控液滴阵列芯片上加工的微结构的形状为凹形,或者凸形,或者平面形结构,作为优选,所述微结构为设置在微流控液滴阵列芯片内的凹槽结构、平面结构、柱状凸起结构;所述微结构中承载液滴的体积范围是I阿升至1_升。 The shape of the microstructures on the microfluidic processing chip array of droplets concave, or convex, or planar configuration, preferably, the microstructure is disposed in a groove microfluidic droplet array chips structure, the planar structure, the columnar convex structure; volume range of the microstructure is carried in droplets raised 1_ a I liter.

[0017]根据所操控液体的性质,利用物理或者化学方法,对所述微结构的表面进行处理,使表面对操控液体具有亲和性。 [0017] The nature of the liquid being manipulated by a physical or chemical method, the microstructure of the surface is treated so that the surface has an affinity for the liquid control. 作为优选,所述微结构表面对被取液体具有亲和性。 Advantageously, the microstructured surface having an affinity for the liquid to be taken.

[0018]作为优选,所述取液端为平面结构或者锥形结构。 [0018] Advantageously, the flat end of a tapping structure or tapered structure. 采用该技术方案,可根据需要取液量,对取液端浸入被取液体中的深度进行调整。 With this aspect, the depth can be adjusted according to the amount of liquid needed to take on the end of the dip is taken to take liquid in the liquid.

[0019]所述多孔板可采用商品化的96孔板,或384孔板,或1536孔板,或其他类型孔板,用于装载和储存被取液体。 [0019] The porous plate 96 may be used commercially, or 384 well plates or 1536 well, or other type of plate, for loading and storing a liquid to be taken. 一种基于液体残留的超微量液滴操控方法,包括: Ultrafine droplet control method based on the amount of residual liquid, comprising:

[0020] (I)移动取液探针或者多孔板,使所述取液探针插入多孔板孔槽内的被取液体中,使取液探针取液端的部分表面或者全部表面浸没到所述被取液体中,蘸取多孔板内的被取液体;取液探针支持部分的表面可不浸入或部分浸入被取液体中; [0020] (I) a tapping move the probe or a porous plate, so that the liquid is taken the liquid take probe insertion hole of the porous plate in the groove, so that part of the end surface of the probe take liquid or extracting solution to the entire surface of the immersed said liquid is taken, is dipped into the liquid in the perforated plate taken; a tapping surface portion may not support the probe immersed or partially immersed in the liquid is taken;

[0021] (2)移动取液探针或者多孔板,使取液探针的取液端脱离多孔板内的被取液体,取液端上表面形成被取液体液滴;该步骤中,少量被取液体残留在取液探针的取液端表面形成液滴,完成取液操作;这一操作也可称为蘸取液体的操作;在所述取液探针浸入被取液体的过程中,取液探针支持部分的表面不残留被取液体,或其残留的被取液体量相对取液端残留的液体量极小,可被忽略; [0021] (2) a tapping move the probe or a porous plate, so that the liquid taken from the end of the probe liquid is taken for liquid within the porous plate, the surface of the liquid droplets is formed on a tapping end is taken; In this step, a small amount of the residue was taken for liquid in the liquid end surface of the probe take liquid form droplets, a tapping operation is completed; this operation may also be called an operating dipped into a liquid; the liquid immersion liquid is taken in the process of taking the probe , the surface of the probe holding part to take liquid is taken the liquid is not left, or the remaining amount of liquid is taken to take the opposite ends of the liquid is extremely small amount of remaining liquid can be ignored;

[0022] (3)移动取液探针或者微流控液滴阵列芯片,使取液探针的取液端表面的被取液体液滴与微流控液滴阵列芯片上的微结构接触,部分被取液体转移到在微流控液滴阵列芯片上微结构内;所述微流控液滴阵列芯片上的微结构顶面覆盖有与被取液体不互溶的油相; [0022] (3) a tapping move the probe or array chip microfluidic droplets, so that a tapping of the end surface of the probe liquid is taken for liquid droplets in contact with the microstructures on the microfluidic chip is an array of droplets, fluid transfer portion is taken into the microstructure in an array of droplets on a microfluidic chip; the microfluidic microstructure surface on chip covered with an array of droplets and the oil is taken immiscible liquid phase;

[0023] (4)移动取液探针或者微流控液滴阵列芯片,使取液探针的取液端脱离微流控液滴阵列芯片,在微流控液滴阵列芯片的微结构内形成被取液体的液滴,完成在所述微流控液滴阵列芯片上生成液滴的操作。 [0023] (4) a tapping move the probe or array chip microfluidic droplets, so that the liquid taken from the end of the probe to take liquid droplet microfluidic array chip, micro droplets within the microfluidic chip array structure forming droplets of the liquid is taken to complete the operation on the generated droplets in microfluidic droplet array chip.

[0024]在进行步骤(I)前,可根据被取液体的性质,预先分别对取液探针的取液端和支持部分的表面进行选择或者处理,使得所述取液端的表面对被取液体具有亲和性,所述支持部分的表面对被取液体不具有亲和性。 [0024] before performing step (the I), can advance the end surface of respectively extracting solution was taken and the support portion of the probe is selected according to the properties to be taken or the processing liquid, so that the surface of the end take liquid is taken liquid has affinity for the surface of said support having no affinity for the liquid is taken. 然后在多孔板的孔槽内加入多种被取液体,被取液体的种类为化学和生物反应、分析和筛选中所需要的样品或者试剂。 Then for the chemical and biological reactions, screening and analysis of sample or reagent required in the pores of the porous plate of the tank is taken with kinds of liquid added, the liquid was taken.

[0025]为实现两种液滴的混合操作,作为优选,所述微结构内预先承载有与被取液体不同的液滴。 [0025] To achieve the mixing operation two kinds of liquid droplets, Advantageously, said microstructure is taken in advance and carrying different liquid droplets. 采用步骤(I)的操作,将取液探针插入多孔板容器内的其他被取液体中,使取液探针取液端的部分表面或者全部表面浸没到所述被取液体中,利用所述取液探针的取液端表面蘸取该容器内的液体;然后根据步骤(3),移动取液探针或者微流控液滴阵列芯片,使取液探针的取液端表面的被取液体液滴与微流控液滴阵列芯片上的微结构上预先承载的液滴接触,所述两个液滴相互混合(融合)形成新的液滴,完成液滴加液操作或者两液滴混合(融合)操作;所述微流控液滴阵列芯片上的微结构上预先承载的液滴可采用本发明上述的操作所形成的,或者采用其他方法在微流控液滴阵列芯片上的微结构上所预先形成的,或者是采用来自另一个取液探针取液端表面上的液滴代替微流控液滴阵列芯片上的微结构上预先承载的液滴进行加液操作或者两液滴混合操作。 Using the step (I) operation, the probe is inserted into the other to take liquid is taken the liquid in the container the perforated plate, so that part of the end surface of the probe was taken to take the entire surface immersed in the liquid or the liquid is taken in by the was taken to take the end surface of the probe was dipped into the liquid in the container; then in accordance with step (3), a tapping movement of the probe, or an array of droplets microfluidic chip, so that a tapping of the end surface of the probe liquid is taken take the liquid droplet and the droplet contacts the microstructures on the microfluidic chip array of droplets carried in advance, the two droplets mixed together (fusion) the formation of new droplets, was added dropwise to complete the two operations or liquid mixed dropwise (fusion) operation; microfluidic droplets on the microstructures on the control of the droplet array chip carrier in advance of the present invention can be formed by the above-described operations, or by other means in an array of droplets on the microfluidic chip the microstructure of the pre-formed, or the use of droplets on the probe from another end surface of a tapping takes place of the liquid droplets on the microfluidic microstructure array of droplets on the control chip carried by dosing a pre-operation or two droplet mixing operation.

[0026]在完成上述操作完成后,移动取液探针或者微流控液滴阵列芯片,使取液探针的取液端脱离微流控液滴阵列芯片的微结构上所形成的新的液滴,在取液探针的取液端表面也残留了所形成的新液滴中的部分液体,完成了从该液滴中的取液操作;完成从液滴中的取液操作还可采用如下的方法,移动取液探针或者微流控液滴阵列芯片,将另一个取液端表面不带有所亲和液体的取液探针或者带有少量空白亲和液体的取液探针,首先插入微流控液滴阵列芯片的微结构上所承载的液滴,然后再脱离该液滴,在所述取液探针的取液端表面残留了该液滴中的部分液体,完成了从该液滴中的取液操作。 [0026] After the above operation is completed, a tapping movement or probe array chip microfluidic droplets, so that the liquid taken from the end of the probe to take on the new liquid microfluidic microstructure array chip droplets are formed droplets, taking the end surface of the liquid probes also take liquid remaining part of the new liquid droplets formed to complete the operation to take liquid from the droplets; complete solution can also be taken from a droplet operation a method is employed, the probe movement to take liquid droplet arrays or microfluidic chip, and the other end face without having to take liquid was taken and the liquid affinity of the probe or probe was taken and a small amount of the liquid with the affinity blank needle, is first inserted into the droplet on the droplet microfluidic microstructure array chip carried, and then the drop off, the remaining portion of the liquid droplets in the extracting solution was taken of the end surface of the probe, It was taken from the completion of the operation of the droplet.

[0027]本发明利用所述取液探针取液端的部分或者全部表面蘸取多孔板内或液滴内的被取液体,进行取液操作;通过减小与被取液体接触的取液探针取液端的的表面面积,减小取液量,取液体积范围是0.1阿升至1_升。 [0027] The present invention utilizes a tapping take liquid probe end portion or all of the inner surface of the dipped porous plate or the liquid droplet is taken to perform a tapping operation; taken by reducing liquid contact with the liquid taken Probe needle into end of the liquid surface area, reducing the amount of extracting solution, extract solution volume was raised to the range of 0.1 a 1_ liters.

[0028]本发明可采用多个取液探针构成取液探针阵列,平行进行多种液体的取液、混合和转移操作,完成高通量的化学和生物反应、分析和筛选。 [0028] The extracting solution can be a plurality of probes constituting the probe array was taken, to take parallel liquid, mixing and more liquid transfer operation, the completion of chemical and biological reactions in high-throughput, screening and analysis of the present invention. 作为优选,所述取液探针阵列中取液探针的数目以及探针的位置与商品化的96孔板,或384孔板,或1536孔板,或其他类型孔板相匹配,实现与现有系统的无缝衔接;作为优选,在进行反应、分析和筛选前,将各种需要的被取液体预先装载并储存至所述多孔板中,简化操作步骤和缩短操作时间;作为优选,所述多孔板上承载有多种被取液体,且多种被取液体浓度或者组成不完全相同。 Advantageously, the pick-up position 96 and the number of commercial liquid solution taken in the probe array probe and a probe or a 384 well plate or a 1536 well, plates or other types match, and achieve seamless existing systems; preferably, the reaction is performed prior to analysis and screening, the liquid needs to be taken pre-loaded and stored to the porous plate, to simplify the steps and to shorten the operation time; Advantageously, the board carries a plurality of porous liquid is taken, and various concentrations of the liquid is taken or not identical composition. 作为优选,所述微流控液滴阵列芯片预先内承载有多种液滴,且多种液滴浓度或者组成不完全相同。 Advantageously, the microfluidic chip is an array of droplets within the pre-loaded with a variety of liquid droplets, and the droplet concentration or more composed not identical. 作为优选,采用取液探针阵列一次性蘸取部分或者全部多孔板内的不同液体,提高取液操作的通量。 Preferably, the probe array was taken using a disposable portion dipped or different from the liquid in the perforated plate of all, the operation was taken increase throughput.

[0029]本发明在进行纳升级体积以下的取液、混合和转移操作时,取液探针、微流控液滴阵列芯片和多孔板的部分或者全部,分别浸入与被取液体不互溶的油相中,或置于封闭体系,或置于高湿度体系,以防止微量液体的蒸发。 [0029] When carrying out the present invention nanoliter take liquid transfer operation and mixing volume or less, a tapping probe array chip microfluidic droplets and some or all of the perforated plate, and were immersed immiscible liquid is taken the oil phase, or placed in a closed system, the system or exposed to high humidity to prevent evaporation of the trace amount of liquid. 即作为优选,所述被取液体置于多孔板中,且被取液体表面覆盖有与被取液体不互溶的油相。 That Advantageously, the perforated plate placed in the liquid is taken, and the fluid is taken with a surface covered with an oil phase immiscible liquid is taken. 此外,油相的使用有助于降低被取液体在取样探针支持部分的吸附和残留,同时,也有利于降低被取液体在取液探针取液端表面的残留体积。 In addition, the oil phase is taken to help reduce the liquid remaining in the sampling probe and the adsorption support portion, but also help to reduce the volume of the residue is taken through the liquid surface of the end of the probe liquid was taken.

[0030]本发明中,所述的取液探针为一次性取液使用,或者多次性取液使用;当取液探针进行多次性取液使用时,在蘸取不同液体之前对取液探针的取样端进行清洗处理,或者在多次性取液操作所产生的交叉污染可忽略的情况下,直接利用取液探针蘸取多种不同液体,而不需对取液探针的取样端进行清洗处理。 [0030] In the present invention, said extracting solution was taken using a disposable probe or a plurality of times of use was taken; when extracting solution was taken of multiple probes used for different liquid prior dipped take liquid sampling probe end cleaning process, or in the case of cross-contamination of a tapping operation several times produced negligible, the probe was dipped directly take a variety of different liquids without the need to take liquid RESEARCH sampling end of the needle cleaning process.

[0031]本发明的优点主要在于:(I)操控液体具有皮升(10—12升)以下,甚至低至飞升到阿升级(10—15到10—18升)的体积精度,有效降低了化学和生物反应和分析中的样品和试剂消耗,节省了实验成本;(2)无需采用注射栗、蠕动栗等驱动部件和毛细管等管路,系统结构大大简化,成本显著降低;(3)系统操作简单、可靠性高、易于实现阵列化和自动化。 [0031] The main advantage of the invention lies in: (I) having the liquid handling picoliter (10-12 liters) or less, even as low volume precision soar upgrade A (10-15 to 10-18 liters), effectively reducing the chemical and biological reactions and the analysis of the sample and reagent consumption, savings in the cost of experiments; (2) without using an injection chestnut, creeping Li like drive member and capillary piping system structure greatly simplified, costs are significantly reduced; (3) system simple operation, high reliability, easy to implement and automate array.

附图说明 BRIEF DESCRIPTION

[0032]图1是实施例1的基于液体残留的超微量液滴操控方法的操控示意图。 [0032] FIG. 1 is a schematic diagram illustrating an embodiment of a droplet operation method based on the amount of liquid remaining in ultra embodiment.

[0033]图2是实施例1的基于液体残留的超微量液滴操控方法的另一种操控示意图。 [0033] FIG 2 is a control method of a droplet amount of ultrafine Example 1 based on the residual liquid schematic diagram illustrating another embodiment.

[0034]图3是实施例1中使用的由不锈钢大头针所加工的取样探针取液端的显微照片侧视图(左)和横截面俯视图(右)。 [0034] FIG. 3 is a diagram of the sample probe pin made of stainless steel used in a processing liquid takes a photomicrograph end side (left) and a top cross-sectional view (right) FIG.

[0035]图4是实施例1实验中生成的荧光素纳混合液滴的荧光显微照片。 [0035] Experimental Example 1 FIG. 4 is generated in the combined droplet fluorescein sodium fluorescence micrographs embodiment.

[0036]图5是实施例2的基于液体残留的超微量液滴操控方法的操控示意图。 [0036] FIG. 5 is a schematic diagram illustrating the method of Example 2 a droplet manipulation based on the amount of liquid remaining in ultra embodiment.

[0037] 上述附图中: [0037] The accompanying drawings in which:

[0038] 1、取液探针;2、取液端;3、支持部分;4、被取液体;5、多孔板;6、微流控液滴阵列芯片;7、微结构;8、油相;9、液滴;1、液滴;11、液滴;12、探针阵列。 [0038] 1, a tapping probe; 2, a tapping terminal; 3, supporting section; 4, the liquid is taken; 5, the perforated plate; 6, microfluidic droplet array chip; 7 microstructure; 8, oil phase; 9, droplet; 1, droplets; 11, drops; 12, probe array.

具体实施方式 detailed description

[0039]下面以具体实施例来对本发明做进一步说明,但本发明的保护范围不限于此。 [0039] The following specific examples further illustrate the present invention, but the scope of the present invention is not limited thereto.

[0040] 实施例1 [0040] Example 1

[0041]图1是实施例1的基于液体残留的超微量液滴操控方法的示意图。 [0041] FIG. 1 is a schematic diagram ultrafine droplet amount control method based on the liquid remaining in Example 1.

[0042]采用不锈钢大头针(直径600微米)作为取液探针I,首先用化学试剂对清洗干净的取液探针I的表面进行整体疏水化处理,然后将取液探针I的尖端在砂纸上打磨,最终得到尖端横截面(直径60微米)为亲水性、其余表面均为疏水性的取液探针1,其中具有亲水性的尖端横截面作为取液探针I的取液端2,取液端2的横截面积半径约为30微米;具有疏水性的取液探针I的表面作为取液探针I的支持部分3(图3)。 [0042] The stainless steel pin (600 microns in diameter) was taken as a probe I, is first cleaned to take liquid surface I of the overall probe hydrophobic treatment with a chemical agent, and the probe tip I was taken in sandpaper on grinding, the tip of the finally obtained cross-section (diameter 60 microns) is hydrophilic, the remaining surfaces are hydrophobic probe 1 was taken, was taken as a probe having a tapping I, the tip end of the hydrophilic cross-section 2, the end cross sectional area of ​​a radius of 2 was taken about 30 micrometers; take liquid surface I of the probe having a hydrophobic liquid as a probe I taken support portion 3 (FIG. 3). 将10—3M(mol/L)荧光素钠溶液作为被取液体4加入商品化的具有384个孔的多孔板5内。 The inner 10-3M (mol / L) sodium fluorescein solution as added to the liquid is taken commercial porous plate 4 having 384 holes 5. 加工玻璃基质的微流控液滴阵列芯片6,微流控液滴阵列芯片6上加工有浅坑作为微结构7承载液滴,将微流控液滴阵列芯片6放入加入矿物油油相8的培养皿内,使油相8浸没微流控液滴阵列芯片6。 Processing the matrix glass microfluidic droplet array chip 6, an array of droplets microfluidic processing chip 6 has a shallow pit microstructure bearing 7 droplets, the droplet array microfluidic chip 6 was added into mineral oily phase petri dish 8, 8 immersion oil phase droplet microfluidic array chip 6. 将取液探针I的支持部分3固定于一支架上,将多孔板5和微流控液滴阵列芯片6固定于三维平移台上。 Probe I was taken support part 3 is fixed to a holder, and the perforated plate 5 microfluidic droplet array chip 6 is fixed to the three-dimensional translation stage.

[0043]取液操作方法:移动平移台,使得取液探针I的取液端2插入多孔板5内的被取液体4中(图1中(2)所示),然后再移动平移台使得取液探针I的取液端2脱离多孔板5内的被取液体4,微量的被取液体4残留于取液探针I的取液端2的表面,形成被取液体4的液滴9(图1中 [0043] a tapping operation: move the translation stage, such that the extracting solution was taken I probe 2 is inserted into the end 4 of the liquid is taken in the perforated plate 5 (Fig. 1 (2)), then the mobile station translation probe I was taken so that the liquid taken from the end of the liquid 2 is taken within the porous plate 4 5, a slight amount of liquid is taken 4 to take the residual liquid was taken probe I end surface 2, a liquid forming liquid is taken 4 9 dropwise (in FIG. 1

(3)所示)(体积约为10皮升),完成对多孔板5内被取液体4的蘸取操作。 (3)) (volume of about 10 picoliters), dipped in the liquid is taken to complete the operation of the porous plate 4 5. 将取液探针I上所形成的液滴9转移到微流控液滴阵列芯片6上的操作方法是:移动平移台,使浸入油相8的微流控液滴阵列芯片6上的特定微结构7与取液探针I上所形成的液滴9相接触(图1中(4)和(5)所示)。 The droplets of the extract solution formed by 9 Probe I was transferred to a method of operating the array of droplets 6 microfluidic chip is: a mobile translation stage, immersed in an oil phase so that the specific array of droplets 6 microfluidic chip 8 microstructure 79 is in contact with the extracting solution droplets formed by the probe I (FIG. 1 (4) and (5)). 再移动平移台,使微流控液滴阵列芯片6与取液探针I脱离,液滴9中的一部分液体,被一次性地转移到微流控液滴阵列芯片6的微结构7内形成新的液滴9,还有部分液滴9残留在取液探针I的取液端2(图1中(6)所示)。 Then move the translation stage so that the droplet microfluidic array chip 6 with Probe I was taken out, a portion of the liquid droplets 9, is transferred to a disposable microfluidic chip is an array of droplets microstructure 6 7 formed the new droplet 9, part of the liquid droplet 9 taken take liquid remaining in the probe 2 I end (in FIG. 1 (6)). 上述操作也可认为是完成了液滴9的分裂操作。 This operation may be considered to be a split completion of the operation of the droplet 9.

[0044]图2是实施例1的又一种液体操作示意图。 [0044] FIG. 2 is a schematic view of still another operation example of the liquid 1 embodiment.

[0045]采用实施例1的又一种液体操作方法是,首先在多孔板5内的被取液体4的上部加入一层适量的矿物油油相8以防止被取液体4的蒸发,移动平移台,使得取液探针I的取液端2插入多孔板5内的被取液体4中(图2中(2)所示),然后再移动平移台使得取液探针I的取液端2脱离多孔板5内的被取液体4,微量的被取液体4残留于取液探针I的取液端2的表面,形成被取液体4的液滴9(图2中(3)所示),完成对多孔板5内被取液体4的蘸取操作。 [0045] Example 1 using yet another method of operating liquid is first upper portion in the porous plate 5 is taken of the liquid 4 was added an appropriate amount of the mineral layer 8 oily phase is taken to prevent evaporation of liquid 4, the translational movement stage, such that the probe I to take liquid take liquid 2 is inserted into the end 4 of the liquid is taken in the perforated plate 5 (Fig. 2 (2)), and then move the translation stage such that a tapping of a tapping end probe I 2 is taken from the liquid in the porous plate 54, a slight amount of liquid is taken take liquid remaining on the surface 4 2 of the end of a tapping probe I, is taken to the liquid droplet 94 is formed (in FIG. 2 (3) shown), dipped in the liquid is taken to complete the operation on the porous plate 4 5. 下一步将取液探针I上所形成的荧光素纳溶液的液滴9与预先在微流控液滴阵列芯片6上微结构7内形成的体积为10纳升的空白溶液的液滴10相混合。 A droplet of a solution of fluorescein sodium was taken on the next step I probe volume 9 is formed in the preformed droplets microfluidic microstructure array chip 6 7 10 nanoliters blank solution 10 mixed. 其操作方法是:移动平移台,使浸入油相8的微流控液滴阵列芯片6上的特定微结构7中的液滴10与取液探针I上所形成的液滴9相接触(图2中(4)和(5)所示),液滴9的液体与液滴10的液体相混合,液滴9与液滴10融合成为一个新的混合液滴U。 Its method of operation are: translation stage moves the microfluidic immersed in an oil phase microstructures on a particular droplet array chip 8 6 10 7 droplets with a droplet on a tapping I probe 9 is formed in contact with ( FIG 2 liquid liquid droplets (4) and (5)), 9 of 10 droplets are mixed, fused to the droplet 10 9 droplet become mixed droplets of a new U. 再移动平移台,使微流控液滴阵列芯片6与取液探针I脱离,液滴11分裂为两个液滴,一个液滴保留在微流控液滴阵列芯片6的微结构7内,还有一个液滴残留在取液探针I的取液端2 (图2 (6))。 Then move the translation stage so that the droplet microfluidic array chip 6 with Probe I was taken out, the droplet 11 splits into two droplets, the droplet is retained within a microfluidic droplet microstructure array chip 6 7 , there is a droplet liquid remaining in the extract solution taken probe I 2 end (FIG. 2 (6)). 保留在微流控液滴阵列芯片6的微结构7内的液滴11的显微荧光照片如图4所示。 Microscopic photograph droplets remain in the phosphor 11 in the microstructure 7 microfluidic droplet array chip 6 is shown in FIG.

[0046]图3是实施例1中使用的由不锈钢大头针所加工的取样探针取液端的显微照片侧视图(左)和横截面俯视图(右)。 [0046] FIG. 3 is a diagram of the sample probe pin made of stainless steel used in a processing liquid takes a photomicrograph end side (left) and a top cross-sectional view (right) FIG.

[0047]图4是实施例1实验中生成的荧光素纳混合液滴的荧光显微照片。 [0047] Experimental Example 1 FIG. 4 is generated in the combined droplet fluorescein sodium fluorescence micrographs embodiment.

[0048] 实施例2 [0048] Example 2

[0049]图5是实施例2的基于液体残留的超微量液滴操控方法的示意图。 [0049] FIG. 5 is a schematic view of a droplet amount of ultra-based manipulation method according to the second liquid remaining embodiments.

[0050]采用ABS塑料加工圆柱形长棒(直径1.8毫米,长度40毫米)作为取液探针I,不需对取液探针I的表面进行表面处理,使用前仅对其进行简单的清洗。 [0050] ABS plastic processing long cylindrical rod (diameter 1.8 mm, length 40 mm) was taken as I probe, the probe does not need to take liquid surface I of the surface treatment, only a simple washing thereof prior to use . 利用取液探针I的圆柱体顶端的圆形横截面作为取液探针I的取液端2,取液探针I的其他部分作为取液探针I的支持部分3。 Probe I was taken using a circular cross-section to the top of the cylinder was taken as the probe was taken 2 I end, the other part was taken as I probe Probe I was taken support portion 3. 将六个取液探针I的另一端或者其支持部分3固定在一块托板上构成一个一维的取样探针阵列12,为与商品化384孔板匹配,相邻的取液探针I之间的间距为4.5毫米(图5)。 The other end of the probe was taken six I, or a supporting portion 3 is fixed on a pallet to form a one-dimensional sampling probe array 12, with the match plate commercialization 384 adjacent a tapping Probe I the spacing between 4.5 mm (FIG. 5). 将10—3M荧光素钠溶液作为被取液体4加入商品化具有384个孔的多孔板5的六个孔内。 The porous plate 10-3M fluorescein sodium is taken as a liquid having added 4 commercialization six holes 384 of the bore 5. 加工聚二甲基硅氧烷(PDMS)基质的微流控液滴阵列芯片6,微流控液滴阵列芯片6上加工有凸起的圆柱形(直径2毫米,高度0.1毫米)作为微结构7承载液滴,将微流控液滴阵列芯片6放入加入矿物油油相8的培养皿内,使油相8浸没微流控液滴阵列芯片6。 Processing polydimethyl siloxane (PDMS) microfluidic droplet matrix array chip 6, an array of droplets microfluidic processing chip 6 on a raised cylindrical (diameter 2 mm, height 0.1 mm), as microstructured 7 carries the droplets, the droplets microfluidic array chip 6 was added into the petri dish with the mineral oil level of 8, 8 immersion oil phase droplet microfluidic array chip 6. 将取样探针阵列12通过其托板固定于一支架上,将多孔板5和微流控液滴阵列芯片6固定于三维平移台上。 The sampling probe array 12 by a frame which is fixed to the pallet, the perforated plate 5 and the microfluidic droplet array chip 6 is fixed to the three-dimensional translation stage.

[0051]液体操纵方法:移动平移台,使得各个取液探针I的取液端2分别插入多孔板5内的对应的被取液体4中(图5(1)),然后再移动平移台使得各个取液探针I的取液端2分别脱离多孔板5内的对应的被取液体4,微量的被取液体4分别残留于各个取液探针I的取液端2的表面,形成多个被取液体4的液滴9(图5中(2)所示)(体积约为800纳升),完成对多孔板5内各个被取液体4的蘸取操作。 [0051] Liquid manipulation methods: moving the translation stage, such that the respective probes I take liquid was taken ends 2 are inserted in the corresponding liquid is taken in the porous plate 54 (FIG. 5 (a)), and then moving the translation stage so that each extracting solution was taken probe I end 2 is taken respectively from the corresponding liquid in the porous plate 54, a slight amount of liquid is taken in the remaining 4 were taken were taken each probe liquid surface I of the end 2, forming a plurality of liquid droplet 94 is taken in (in FIG. 5 (2)) (volume of about 800 nanoliters), to complete the respective operation dipped porous plate 5 is taken within the liquid 4. 下一步将取液探针I上所形成的液滴9与预先在微流控液滴阵列芯片6上微结构7内形成的体积为800纳升的另一溶液的液滴10分别一一对应混合。 The next step was taken on droplets formed by the probe 9 and I volume of pre-formed droplets in the microfluidic microstructure array chip 6 7 800 nanoliter droplets 10 respectively correspond to another solution mixing. 其操作方法是:移动平移台,使浸入油相8的微流控液滴阵列芯片6上的特定微结构7上的各个液滴1与取液探针I上所形成的各个液滴9 一一对应相接触(图5中(3)所示),液滴9的液体与液滴1的液体相混合,液滴9与液滴1融合成为一个新的混合液滴11 (图5中(4)所示)。 Its method of operation are: translation stage moves the microfluidic immersed in an oil phase droplets each individual droplets 1 droplet 78 on a particular array of microstructures on the chip 6 and the probe was taken I formed a 9 corresponding to a contact (in FIG. 5 (3)), the liquid 9 with a droplet of the liquid 1 are mixed, droplets 1 droplet 9 become fused to a new mixed droplet 11 ((FIG. 5 shown) 4). 再移动平移台,使微流控液滴阵列芯片6与各个取液探针I脱离,每个液滴11分裂为两个液滴,一个液滴保留在微流控液滴阵列芯片6和微结构7内,还有一个液滴残留在取液探针I的取液端2(图5中(5)所示)。 Then move the translation stage so that the droplet microfluidic array chip 6 and the respective probes I was taken out, each droplet 11 is split into two droplets, the droplets remain in a droplet microfluidic array chips and micro 6 within the structure 7, and a liquid droplet remaining on the probe is taken to a tapping terminal I 2 (FIG. 5 (5)).

[0052]构成六个液滴9的溶液的组成可以是相同的,以进行同一样品或试剂的反应,分析和筛选;它们的组成也可以是不同的,以进行多种不同的样品或者试剂的反应,分析和筛选。 [0052] The solution composition composed of six droplets 9 may be the same, to carry out the reaction in the same sample or reagents, and screening analysis; thereof may also be different, for a plurality of different samples or reagents reaction, analysis and screening. 同样地,构成微流控液滴阵列芯片6六个液滴10的溶液的组成也可以是相同的,以进行同一样品或试剂的反应,分析和筛选;它们的组成也可以是不同的,以进行多种不同的样品或者试剂的反应,分析和筛选。 Likewise, the composition of the solution constituting the microfluidic droplet array chip 6 six droplets 10 may be the same, for reaction in the same sample or reagents, and screening analysis; thereof may also be different to the reaction of various different samples or reagents, analysis and screening.

Claims (9)

  1. 1.一种基于液体残留的超微量液滴操控装置,其特征在于,包括: 多孔板,该多孔板上设有一个或多个用于盛放被取液体的孔槽; 微流控液滴阵列芯片,该微流控液滴阵列芯片上设有一个或多个用于承载液滴的微结构; 取液探针,该取液探针上具有支持部分、以及位于支持部分底端的取液端;所述取液端表面对被取液体的亲和性大于等于所述支持部分外表面对被取液体的亲和性;所述取液探针的外形为实心针形或实心柱形;所述取液端表面的面积为100平方纳米至1000平方毫米; 切换机构,依次实现取液探针对被取液体的取样操作以及将所取被取液体置于微流控液滴阵列芯片内的放样操作。 A droplet amount control means based ultra residual liquid, characterized by comprising: a porous plate, the porous plate provided with a plurality of holes or slots for accommodating a liquid to be taken; microfluidic droplets array chips on the array chip microfluidic droplets provided with one or more microstructures for carrying droplets; a tapping probe supporting portion having a tapping on the probe, and located to support the bottom end portion of the liquid taken end; the end surface of the liquid to take the affinity for liquid is greater than or equal to the outer face of the support section is taken affinity for the liquid; the liquid takes the shape of a solid probe or needle-shaped solid cylindrical; the end surface of the liquid takes an area of ​​100 square nanometers to 1000 mm2; switch mechanism sequentially to achieve a tapping operation on the sampling probe and the liquid is taken to be taken for liquid droplets disposed within a microfluidic array chips the stake-out operation.
  2. 2.根据权利要求1所述的基于液体残留的超微量液滴操控装置,其特征在于,所述微结构为设置在微流控液滴阵列芯片内的凹槽结构、平面结构、柱状凸起结构;所述微结构中承载液滴的体积范围是I阿升至I毫升。 The ultra trace residual liquid droplet-based manipulation apparatus according to claim 1, wherein said microstructures disposed within a groove structure in an array of droplets microfluidic chip, a planar structure, a columnar protrusion structure; the volume range of the microstructure is carried in droplets rose I a I ml.
  3. 3.根据权利要求1所述的基于液体残留的超微量液滴操控装置,其特征在于,所述微结构表面对被取液体具有亲和性。 According to claim an ultrafine liquid droplets remaining amount control apparatus of claim 1, wherein said microstructured surface having an affinity for the liquid to be taken.
  4. 4.根据权利要求1所述的基于液体残留的超微量液滴操控装置,其特征在于,所述取液端为平面结构、或者柱形结构、或者锥形结构。 The residual liquid ultrafine droplet amount based on the control apparatus of claim 1, wherein said planar end structure was taken, or columnar structure, or tapered structure.
  5. 5.—种利用权利要求1-4任一权利要求所述的装置进行超微量液滴操控方法,其特征在于,包括: (1)使取液探针取液端的部分表面或者全部表面浸没到多孔板内的被取液体中,蘸取多孔板内的被取液体,被取液体粘附于取液探针取液端; (2)使取液探针的取液端脱离多孔板内的被取液体,取液端上表面形成被取液体液滴; (3)使取液探针的取液端表面的被取液体液滴与微流控液滴阵列芯片上的微结构接触,或者当微结构上预先承载有液滴时,与该液滴接触,部分被取液体转移到在微流控液滴阵列芯片上微结构内,或者与微结构上预先承载的液滴混合;所述微流控液滴阵列芯片上的微结构顶面覆盖有与被取液体不互溶的油相; (4)使取液探针的取液端脱离微流控液滴阵列芯片,在微流控液滴阵列芯片的微结构内形成被取液体的液滴,完成在所述微流控液滴阵 The device according to any one of claims 1-4 5.- species for use as claimed in claim ultrafine droplet amount control method comprising: (1) so that part of the surface of the probe end of the extracting solution was taken, or to the entire surface submerged is taken in the liquid in the perforated plate, porous plate is dipped for liquid, the liquid is taken adhered to the extract solution was taken probe end; (2) reacting a tapping end of the probe was taken from the porous plate is taken the liquid, the liquid droplets formed on the surface is taken to the end of a tapping; is taken to be the liquid droplets (3) so that an end surface of a tapping take liquid microstructures probe contacts, an array of droplets on the microfluidic chip or when the microstructured previously carried droplet in contact with the drop, the liquid portion is transferred to take on a microstructure microfluidic droplet array chip, or mixed with droplets carried by the microstructured beforehand; the microstructure surface covering the droplets on the microfluidic chip array is taken with oil immiscible liquid phase; (4) was taken from the end of the probe to take liquid droplet microfluidic array chip, microfluidic for liquid droplets are formed in the microstructure of the droplet array chips, to complete the microfluidic droplet array 芯片上生成液滴的操作; 任选进入步骤(5)和(6): (5)选择另一个取液探针,使取液探针取液端的部分表面或者全部表面浸没到流控液滴阵列芯片上生成液滴中,蘸取液滴,部分液滴粘附于取液探针取液端; (6)使取液探针脱离的取液端脱离微流控液滴阵列芯片,完成部分液滴的取液操作。 Generating a droplet operation on the chip; optionally proceeds to step (5) and (6): (5) selecting a tapping another probe, so that part of the end surface of the probe was taken to take the entire surface immersed in liquid or droplet flow control generating droplets of the array chips, dipped into droplets, the droplets adhered to a tapping portion probe end extracting solution; (6) so that the liquid taken from the end of the probe was taken from the microfluidic droplet array chip is completed operating portion to take liquid droplets.
  6. 6.根据权利要求5所述的进行超微量液滴操控方法,其特征在于,所述被取液体置于多孔板中,且被取液体表面覆盖有与被取液体不互溶的油相。 According to claim 5, wherein the amount of liquid droplets is ultra control method, characterized in that the liquid is taken perforated plate placed in, and is covered with the oil phase has taken for liquid immiscible liquid surface.
  7. 7.根据权利要求5所述的进行超微量液滴操控方法,其特征在于,所述微结构内预先承载有与被取液体不同的液滴。 According to claim 5, wherein the amount of liquid droplets is ultra control method, wherein said microstructure is taken with pre-loaded with a different liquid droplets.
  8. 8.根据权利要求5所述的进行超微量液滴操控方法,其特征在于,所述多孔板上承载有多种被取液体,且多种被取液体浓度或者组成不完全相同。 According to claim 5, wherein the amount of liquid droplets is ultra control method, wherein the porous plate has a plurality of carrier liquid is taken, and various concentrations of the liquid is taken or not identical composition.
  9. 9.根据权利要求7所述的进行超微量液滴操控方法,其特征在于,所述微流控液滴阵列芯片预先内承载有多种液滴,且多种液滴浓度或者组成不完全相同。 According to claim 7 ultrafine droplet amount control method, wherein the microfluidic chip is an array of droplets within the pre-loaded with a variety of liquid droplets, and the droplet concentration or composition is not more identical .
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