CN101474541B - Integrated chip and device thereof, and method for preparing micrometre level dispersoid - Google Patents

Abstract

The invention discloses an integrated chip, a device adopting the integrated chip and a method for preparing micron-sized dispersoid; the substrate of the integrated chip is the substrate with hydrophilic surface; two layers of adjacent structures are a group of structures, and the integrated chip comprises at least one group of structures; in one group of structures, the first structure is provided with M levels of gradients, and the second structure is provided with N levels of gradients; wherein, M and N are natural numbers, and M is less than N; among the gradients, the primary gradient is provided with H1 channel 1, H2 branches are led out from each channel 1 in the secondary gradient, and H1*H2...*HN channel N are formed; one side which is far away from each channel M is provided with a connecting channel respectively; the integrated chip also comprises at least a collecting groove; the integrated chip also comprises at least one collecting groove which is connected with the output end of each collecting channel for collecting and exporting products. The invention has very high productivity and absolute advantage in the aspect of the large scale production of contrast agent.

Description

Translated fromChinese

集成芯片及其装置、以及制备微米级分散体的方法 Integrated chip and device thereof, and method for preparing micron-scale dispersion

【技术领域】【Technical field】

本发明涉及应用造影剂对比显像技术，尤其涉及的是，一种采用微流控技术制造微米级分散体的大规模集成芯片；一种采用该芯片的装置，可用于制备超声造影剂；以及采用该装置制备微米级分散体的方法，微米级分散体可以用作超声造影剂。 The present invention relates to the application of contrast agent contrast imaging technology, in particular, a large-scale integrated chip using microfluidic technology to manufacture micron-scale dispersions; a device using the chip, which can be used to prepare ultrasonic contrast agents; and The method for preparing the micron-scale dispersion by using the device can be used as an ultrasound contrast agent. the

【背景技术】【Background technique】

在临床诊断中，无创、实时、动态地检测有关组织的结构和功能信息已经变得日益重要。超声诊断技术不仅具有上述的特点，而且，与X射线、核磁共振成像技术相比，还具有安全、适应面广、可反复检查、对软组织的鉴别能力强、灵活性强及价格低廉等优点，是世界上使用最广泛的影像技术。 Noninvasive, real-time, and dynamic detection of structural and functional information about tissues has become increasingly important in clinical diagnosis. Ultrasonic diagnostic technology not only has the above-mentioned characteristics, but also has the advantages of safety, wide adaptability, repeated inspection, strong ability to identify soft tissues, strong flexibility and low price compared with X-ray and MRI technologies. It is the most widely used imaging technology in the world. the

应用造影剂对比显像是影像技术发展的核心内容。与各种影像技术相比，超声是应用范围最广泛但惟一未能常规应用造影剂的影像技术，对比超声技术的核心是超声造影剂。 The application of contrast agent contrast imaging is the core content of the development of imaging technology. Compared with various imaging techniques, ultrasound is the most widely used imaging technique but the only imaging technique that does not routinely use contrast agents. The core of contrast ultrasound technology is ultrasound contrast agents. the

1968年，Gramiak等首次观察到经导管注射含气盐水可使右心显影增强，揭开了心脏声学造影的序幕。在这一阶段，人们主要利用通过手振生理盐水或CO2发泡剂等方法制作超声造影剂。其微泡直径较大、均一性不佳，难以通过肺循环。另外因为无外壳的保护，微泡中气体迅速在血液中弥散使得微泡存活时间短暂，有学者将这一时期的造影剂称为第0代超声造影剂。 In 1968, Gramiak et al. observed for the first time that transcatheter injection of gas-containing saline could enhance the imaging of the right heart, which opened the prelude to echocardiography. At this stage, people mainly use methods such as vibrating physiological saline or CO2 foaming agent to make ultrasound contrast agents. Its microbubbles are large in diameter and poor in uniformity, making it difficult to pass through the pulmonary circulation. In addition, because there is no protection of the shell, the gas in the microbubbles diffuses rapidly in the blood, making the microbubbles survive for a short time. Some scholars call the contrast agent in this period the 0th generation ultrasound contrast agent. the

1984年Feinstein等采用声振法制备得到稳定的微气泡才真正进入左心造影时代。最开始出现的是包裹空气的微泡，包裹空气的外壳材料种类繁多，大致可以分为：蛋白质外壳、脂质体外壳、表面活性剂外壳和高聚物外壳等，称为第一代超声造影剂。由于空气在血液中的溶解度和弥散度较大，微泡内的空气容易通过外壳弥散入血液，使得造影剂微泡变小或消失。另外，因为微泡的散射信号与直径的6次方成正比，只要有少量的空气溢出，微泡的直径轻度变小，就会使微泡的回声信号明显衰减。具有代表性的产品有美国的Albunex、德国Schering公司的Levovist，第一代的超声造影剂的稳定性和存活时间不够长，极大的限制了它的应用，2000年后已逐渐退出造影剂市场及研究领域。 In 1984, Feinstein et al. prepared stable microbubbles by the vibroacoustic method and really entered the era of left heart angiography. The microbubbles that wrap the air first appeared, and there are many types of shell materials that wrap the air, which can be roughly divided into: protein shells, liposome shells, surfactant shells, and polymer shells, which are called the first generation of contrast-enhanced ultrasound. agent. Due to the high solubility and dispersibility of air in the blood, the air in the microbubbles can easily diffuse into the blood through the shell, so that the microbubbles of the contrast agent become smaller or disappear. In addition, because the scattering signal of the microbubble is proportional to the sixth power of the diameter, as long as a small amount of air overflows and the diameter of the microbubble becomes slightly smaller, the echo signal of the microbubble will be significantly attenuated. Representative products include Albunex from the United States and Levovist from Schering in Germany. The stability and survival time of the first generation of ultrasound contrast agents are not long enough, which greatly limits its application. After 2000, it has gradually withdrawn from the contrast agent market. and research areas. the

从1995年以来，因为高分子量、低血液溶解度和弥散性的氟碳氟硫类气体的引入，使超声造影剂的存活时间及稳定性有了显著提高，称为第二代超声造影剂。具有代表性的超声造影剂美国的Optison、 EchoGen和AI-700等，意大利Bracco公司的SonoVue(BR1)和BR14等。 Since 1995, the survival time and stability of ultrasound contrast agents have been significantly improved due to the introduction of high molecular weight, low blood solubility and dispersibility of fluorocarbon, fluorine and sulfur gases, which are called second-generation ultrasound contrast agents. Representative ultrasound contrast agents such as Optison in the United States, EchoGen and AI-700, etc., SonoVue (BR1) and BR14 from Bracco, Italy.

目前，国外已有多种新一代产品上市，而国内没有自主上市的产品，意大利Bracco公司生产的是唯一被我国卫生部批准商品化的超声造影剂，价格十分昂贵。 At present, a variety of new-generation products have been launched abroad, but there is no domestically launched product. The only ultrasound contrast agent produced by the Italian company Bracco has been approved for commercialization by the Ministry of Health of my country, and the price is very expensive. the

总而言之，常规超声造影剂是一种含高浓度微泡的悬浮液，微泡以白蛋白、脂质、表面活性剂或高聚物为外壳，平均直径约2-4微米，小于红细胞，经周围静脉注射后能够稳定的通过肺循环到达左心系统，进而抵达全身各器官。超声造影剂微泡与血液及人体组织的声阻抗差别极大，与超声波之间的作用非常复杂，可以改变所在部位的背向散射系数、衰减系数、声速及其它声学特性，注入血管后能增强局部组织的超声回波能力，从而明显改善超声影像对组织血流灌注的显影效果，显著提高超声诊断的敏感性和特异性，突破了常规超声发展的瓶颈，进一步拓展超声诊断领域。 All in all, the conventional ultrasound contrast agent is a suspension containing a high concentration of microbubbles. The microbubbles are shelled with albumin, lipids, surfactants or polymers, with an average diameter of about 2-4 microns, smaller than red blood cells, and passed through the surrounding area. After intravenous injection, it can stably reach the left heart system through the pulmonary circulation, and then reach various organs of the whole body. The acoustic impedance of ultrasound contrast agent microbubbles is very different from that of blood and human tissue, and the interaction with ultrasound is very complicated. It can change the backscatter coefficient, attenuation coefficient, sound velocity and other acoustic characteristics of the part, and it can enhance the blood pressure after being injected into the blood vessel. The ultrasonic echo capability of local tissues can significantly improve the imaging effect of ultrasonic images on tissue blood perfusion, significantly improve the sensitivity and specificity of ultrasonic diagnosis, break through the bottleneck of conventional ultrasonic development, and further expand the field of ultrasonic diagnosis. the

并且，超声造影剂不仅可用于常规造影增强，还可用于靶向诊断和介入治疗，将特异性配体结合到造影剂微泡表面，使它到达感兴趣的组织或器官，选择性地与相应受体结合，从而达到特异性增强靶区超声信号或局部靶向治疗的目的。而介入治疗的研究，使超声造影剂的应用范围不断扩大，应用价值不断提升。 Moreover, ultrasound contrast agents can be used not only for conventional contrast enhancement, but also for targeted diagnosis and interventional therapy, by binding specific ligands to the surface of contrast agent microbubbles, allowing them to reach tissues or organs of interest, and selectively interacting with corresponding Receptor binding, so as to achieve the purpose of specifically enhancing the ultrasound signal of the target area or local targeted therapy. The research on interventional therapy has continuously expanded the application range of ultrasound contrast agents and continuously improved their application value. the

例如，利用造影剂微泡降低超声空化阈值，从而促进超声溶栓或提高高功率聚焦超声的治疗效果；或利用超声波破坏携带基因或药物的造影剂微泡，使基因或药物靶向释放并促进其穿透，以介导基因转移或药物靶向治疗；或利用造影引导射频或微波治疗并即时评价治疗效果等。 For example, using contrast agent microbubbles to reduce the ultrasound cavitation threshold, thereby promoting ultrasonic thrombolysis or improving the therapeutic effect of high-power focused ultrasound; or using ultrasound to destroy contrast agent microbubbles carrying genes or drugs, so that genes or drugs are released and Promote its penetration to mediate gene transfer or drug-targeted therapy; or use imaging to guide radiofrequency or microwave therapy and evaluate the therapeutic effect in real time. the

靶向超声造影剂的研究正成为超声影像学领域的热点。无论从活性基团的选择、修饰方法的发明、靶向作用的检测，还是体内药代药效研究和临床应用都取得了非常可喜的成绩。随着高分子材料的快速发展和微泡制备工艺的不断完善，微泡超声造影剂必将朝着更加个性化的方向发展，未来的微泡超声造影剂不仅能应用于不同生理和病理状态下的特异超声成像，还可以作为携带药物或治疗基因的载体，在治疗领域开辟一片新的天地。为此，国外医疗机构预测 超声造影剂不久将成为超声诊断或治疗中的常用试剂，具有巨大的社会效益和经济价值。 The research on targeted ultrasound contrast agents is becoming a hot spot in the field of ultrasound imaging. Very gratifying achievements have been made in terms of the selection of active groups, the invention of modification methods, the detection of targeting effects, and the research of pharmacokinetics and pharmacodynamics in vivo and clinical applications. With the rapid development of polymer materials and the continuous improvement of microbubble preparation technology, microbubble ultrasound contrast agents will definitely develop in a more personalized direction. Future microbubble ultrasound contrast agents can not only be used in different physiological and pathological conditions The specific ultrasound imaging can also be used as a carrier to carry drugs or therapeutic genes, opening up a new world in the field of treatment. For this reason, foreign medical institutions predict that ultrasound contrast agents will soon become a common reagent in ultrasound diagnosis or treatment, with huge social and economic value. the

目前，超声领域研究的热点集中在两方面：一是各种增强超声造影的成像技术，如：触发成像技术、二次谐波成像技术、次谐波成像技术、基波脉冲抵消成像技术、能量脉冲反向成像技术、低机械指数实时成像技术等；另一方面则是研究新的更加稳定可靠的超声造影剂。 At present, the research hotspots in the field of ultrasound are concentrated in two aspects: one is various imaging technologies for enhanced contrast-enhanced ultrasound, such as: triggered imaging technology, second harmonic imaging technology, sub-harmonic imaging technology, fundamental wave pulse offset imaging technology, energy Pulse reverse imaging technology, low mechanical index real-time imaging technology, etc.; on the other hand, research on new more stable and reliable ultrasound contrast agents. the

理想的超声造影剂应具备以下特性：(1)安全无毒，粘稠度低，无生物活性，对循环系统没有生理上的影响，不明显稀释红细胞浓度；(2)微泡大小符合要求，具有一定的气泡压力，使灌注后保持足够长的时间和足够强的回波强度；(3)反射性好，衰减伪差小，回波信号及多普勒信号增强明显，能够发生谐波共振增大背散射面积；(4)能静脉给药，体内稳定性好，使用方便，对造影显像的仪器条件没有极为特殊的要求，便于普及；(5)具有稳定的浓度和剂型，易于消毒、灭菌、保存、运输，能够批量生产。 An ideal ultrasound contrast agent should have the following characteristics: (1) safe and non-toxic, low viscosity, no biological activity, no physiological impact on the circulatory system, and no significant dilution of red blood cell concentration; (2) microbubble size meets the requirements, It has a certain bubble pressure, so that it can maintain a long enough time and a strong enough echo intensity after perfusion; (3) Good reflectivity, small attenuation artifact, obvious enhancement of echo signal and Doppler signal, and harmonic resonance can occur Increase the backscattering area; (4) It can be administered intravenously, has good stability in the body, is easy to use, and has no special requirements for contrast imaging equipment, and is easy to popularize; (5) Has a stable concentration and dosage form, and is easy to sterilize , Sterilization, preservation, transportation, capable of mass production. the

目前最常用的制备超声造影剂微泡的方法是声振法和机械振荡法。 At present, the most commonly used methods for preparing ultrasound contrast agent microbubbles are acoustic vibration method and mechanical oscillation method. the

声振法是利用超声波振荡时产生的高频变换的正负声压，其中的负声压使存在于造影剂制备液中的气体膨胀形成微小气泡，此时，制备液中的脂质或白蛋白、表面活性剂、多聚体等趁机包裹微小气泡形成稳定的造影剂微泡。超声造影剂中的白蛋白类、脂类、多聚体类、表面活性剂类等，其制备过程都是采用声振法。 Acoustic vibration method is to use the positive and negative sound pressure of high-frequency transformation generated during ultrasonic oscillation, and the negative sound pressure in it makes the gas in the contrast agent preparation solution expand to form tiny bubbles. At this time, the lipid or white in the preparation solution Proteins, surfactants, polymers, etc. take the opportunity to wrap microbubbles to form stable contrast agent microbubbles. Albumins, lipids, polymers, surfactants, etc. in ultrasound contrast agents are all prepared by the sono-vibration method. the

但是，声振法存在以下缺点：(1)探头式声振仪的工艺参数，包括功率、探头在液面的位置、深度等，不易控制，工艺重现性受到一定影响；(2)由于探头要放在制备液内部，因此在声振过程中很难做到无菌操作，并存在金属污染的可能，给造影剂的质量控制、制备工艺增加了一定难度；(3)声振过程中产生较多的热量，对脂质的活性，尤其在制备携带某些配体、药物或基因的造影剂时，对同时声振的配体、药物或基因的活性产生很大的影响。 However, the vibroacoustic method has the following disadvantages: (1) The process parameters of the probe-type vibro-acoustic instrument, including power, position of the probe on the liquid surface, depth, etc., are not easy to control, and the process reproducibility is affected to a certain extent; (2) due to the It must be placed inside the preparation solution, so it is difficult to perform aseptic operation during the acoustic vibration process, and there is a possibility of metal contamination, which adds a certain degree of difficulty to the quality control and preparation process of the contrast agent; (3) More heat has a great impact on the activity of lipids, especially when preparing contrast agents carrying certain ligands, drugs or genes, and the activity of ligands, drugs or genes that are vibrated simultaneously. the

机械振荡法是利用高频机械振荡时，制备液中各点受力的时相不同产生不同的正负压力，其中的负压可使存在于制备液中的气体形成微小气泡。频率越高，正负压力的变换越快，负压的时间越短，气体膨胀越小，形成的气泡就越小；振幅越小，产生的负压力越小，相同时间情况下，由负压产生的气体膨胀越小，形成的气泡越小，由于制备超声造影剂需要形成较小的微泡，因此，制 备超声造影剂的机械振荡装置需要较高的频率、较低的振幅。 The mechanical oscillation method uses high-frequency mechanical oscillation to produce different positive and negative pressures in different phases of the stress on each point in the preparation liquid, and the negative pressure can make the gas in the preparation liquid form tiny bubbles. The higher the frequency, the faster the positive and negative pressure changes, the shorter the negative pressure time, the smaller the gas expansion, and the smaller the bubbles formed; the smaller the amplitude, the smaller the negative pressure generated. The smaller the expansion of the generated gas, the smaller the bubbles formed. Since the preparation of ultrasound contrast agents requires the formation of smaller microbubbles, the mechanical oscillation device for the preparation of ultrasound contrast agents requires a higher frequency and lower amplitude. the

但是，机械振荡法存在以下缺点：(1)造影剂微泡粒径大小不能够精确控制；(2)形成的微泡粒径分布比较宽，声学特性不稳定，微泡中均含有一定数量大于10微米的微泡，这些大的微泡可能会造成局部血管的堵塞或者破裂，使得超声造影剂的使用潜存一定的风险性；(3)形成微泡的外壳厚度不均匀；(4)机械振荡方法来产生足够的力把气体从周围环境引入到液体溶液中形成气泡，振荡速度很大程度上决定形成微泡的数量和大小，如国内临床批准使用的造影剂SonoVue，使用手摇的方式制备，重复性不够好。 However, the mechanical oscillation method has the following disadvantages: (1) the particle size of the microbubbles of the contrast agent cannot be precisely controlled; (2) the particle size distribution of the formed microbubbles is relatively wide, and the acoustic properties are unstable. Microbubbles of 10 microns, these large microbubbles may cause blockage or rupture of local blood vessels, making the use of ultrasound contrast agents potentially risky; (3) the shell thickness of the microbubbles is not uniform; (4) mechanical Oscillation method is used to generate sufficient force to introduce gas from the surrounding environment into the liquid solution to form bubbles. The oscillation speed largely determines the number and size of microbubbles formed. For example, SonoVue, a contrast agent approved for clinical use in China, uses a manual method preparation, the reproducibility is not good enough. the

因此，现有技术存在缺陷，需要改进。 Therefore, there are defects in the prior art and need to be improved. the

【发明内容】【Content of invention】

本发明所要解决的技术问题是提供一种实用的大规模微流控流动聚焦装置，以及粒径均一、可控制、单分散的声学性质稳定的超声造影剂制备方法，以制备粒径大小可控并且呈单分散性的超声造影剂。在此基础上，提供一种集成芯片、采用该集成芯片的装置和采用该装置制备微米级分散体的方法。 The technical problem to be solved by the present invention is to provide a practical large-scale microfluidic flow focusing device, and a method for preparing ultrasonic contrast agents with uniform particle size, controllability, monodisperse acoustic properties, and controllable particle size. And it is a monodisperse ultrasound contrast agent. On this basis, an integrated chip, a device using the integrated chip and a method for preparing micron-scale dispersions using the device are provided. the

本发明的技术方案如下： Technical scheme of the present invention is as follows:

集成芯片的一种技术方案是，以两层相邻结构为一组结构，所述集成芯片包括至少一组结构；在一组结构中，第一层结构设置M级梯度，第二层结构设置N级梯度；其中，M、N是自然数，M小于N；各级梯度中，第一级梯度设置H1个通道1，第二级梯度从每个通道1中引出H2个分支，形成H1×H2个通道2，以此类推，第N级梯度从每个通道(N-1)中引出HN个分支，形成H1×H2......×HN个通道N； A technical scheme of an integrated chip is to use two layers of adjacent structures as a group of structures, and the integrated chip includes at least one group of structures; in one group of structures, the first layer structure is provided with M-level gradients, and the second layer structure is provided with N-level gradients; among them, M and N are natural numbers, and M is less than N; among the gradients at all levels, the first-level gradient setsH1 channels 1, and the second-level gradient leads to H2 branches from eachchannel 1, forming H1×H2 channel 2, and so on, the Nth level gradient leads to HN branches from each channel (N-1), forming H1×H2...×HN channels N;

在一组结构中，对于第一层结构的任一特定通道M，均对应于第二层结构从一通道M所引出的在第N级梯度形成H(M+1)×H(M+2)......×H(N-1)×HN数量的特定通道N； In a group of structures, for any specific channel M of the first layer structure, it corresponds to the N-th level gradient formed by the second layer structure drawn from a channel M H(M+1)×H(M+2 )......×H(N-1)×HN number of specific channels N; 

其中，各特定通道N的出口，等距分布在一预设置半径圆形的圆周；对于该特定通道M，其输出方向垂直于该圆形，其出口与该圆形的垂线通过该圆形的圆心，该特定通道M的出口与该圆心距离一特定长度；各特定通道N的输出方向与所述垂线的夹角为相同锐角或直角；沿该特定通道M的输出方向，在该圆形远离该特定通道M的一侧，设置一收集通道；所述集成芯片还包括至少一与各收集通道的输出端相连接的收集槽，用于收集和导出产物。Wherein, the outlets of each specific channel N are equidistantly distributed on the circumference of a circle with a preset radius; for the specific channel M, its output direction is perpendicular to the circle, and the perpendicular line between the outlet and the circle passes through the circle The center of the circle, the outlet of the specific channel M is a certain distance from the center of the circle; the angle between the output direction of each specific channel N and the vertical line is the same acute angle or right angle; along the output direction of the specific channel M, in the circle A collection channel is provided on the side away from the specific channel M; the integrated chip also includes at least one collection tank connected to the output end of each collection channel for collecting and exporting products.

集成芯片的又一种技术方案是，M＝N-1。 Yet another technical solution of the integrated chip is that M=N-1. the

集成芯片的又一种技术方案是，各特定通道N的输出方向通过该圆形圆心。 Another technical solution of the integrated chip is that the output direction of each specific channel N passes through the center of the circle. the

集成芯片的又一种技术方案是，通道M和通道N的横截面为矩形；所述矩形高度为20μm至30μm、宽度分别为30μm至50μm、50μm至100μm，或者，所述通道M和通道N的横截面矩形宽度分别为50μm至100μm、30μm至50μm。 Another technical scheme of the integrated chip is that the cross-sections of the channel M and the channel N are rectangular; The widths of the cross-sectional rectangles are 50 μm to 100 μm and 30 μm to 50 μm, respectively. the

集成芯片的又一种技术方案是，所述收集通道的结构至少包括以下结构其中之一：外向放大的锥体、球体、半球体或锥台体，其输入端的管径为7μm至25μm，高度为20μm至30μm；或者，圆柱体或长方体，其沿对应的特定通道M输出方向的截面积略小于该特定通道M沿该方向的截面积，高度为20μm至30μm。 Yet another technical solution of the integrated chip is that the structure of the collection channel includes at least one of the following structures: outwardly enlarged cones, spheres, hemispheres or frustums, the diameter of the input end of which is 7 μm to 25 μm, and the height 20 μm to 30 μm; or, a cylinder or a cuboid whose cross-sectional area along the output direction of the corresponding specific channel M is slightly smaller than the cross-sectional area of the specific channel M along this direction, and a height of 20 μm to 30 μm. the

集成芯片的又一种技术方案是，其基质为表面亲水性基质。 Yet another technical solution of the integrated chip is that its substrate is a surface hydrophilic substrate. the

集成芯片的又一种技术方案是，其基质为硅、玻璃、聚二甲基硅氧烷、聚甲基丙烯酸甲酯或聚碳酯。 Yet another technical solution of the integrated chip is that its substrate is silicon, glass, polydimethylsiloxane, polymethylmethacrylate or polycarbonate. the

集成芯片的又一种技术方案是，第一层结构与第二层结构的组合，为气体通道结构与液体通道结构的组合。 Another technical solution of the integrated chip is that the combination of the first layer structure and the second layer structure is a combination of a gas channel structure and a liquid channel structure. the

装置的一种技术方案是，其包括：第一物质输入单元、第二物质输入单元、至少一集成芯片形成的集成芯片阵列、贮存单元；其中，集成芯片为上述任一关于集成芯片的技术方案，所对应的集成芯片； A technical solution of the device is that it includes: a first material input unit, a second material input unit, an integrated chip array formed by at least one integrated chip, and a storage unit; wherein the integrated chip is any of the above technical solutions related to integrated chips , the corresponding integrated chip;

各集成芯片的收集槽，分别与所述贮存单元相连接，用于收集和导出产物；所述第一物质输入单元分别与各集成芯片的一层结构的各通道1相连接，用于输入第一物质；所述第二物质输入单元分别从各集成芯片的另一层结构的各通道1相连接，用于输入第二物质。 The collection tanks of each integrated chip are respectively connected with the storage unit for collecting and exporting products; the first substance input unit is respectively connected with eachchannel 1 of the one-layer structure of each integrated chip for inputting the first substance A substance; the second substance input unit is respectively connected to eachchannel 1 of another layer structure of each integrated chip for inputting the second substance. the

装置的又一种技术方案是，所述第一物质输入单元与所述第二物质输入单元的组合，为气体输入单元与液体输入单元的组合；或者，所述第一物质输入单元与所述第二物质输入单元均为液体输入单元。 Another technical solution of the device is that the combination of the first substance input unit and the second substance input unit is a combination of a gas input unit and a liquid input unit; or, the combination of the first substance input unit and the The second substance input units are all liquid input units. the

装置的又一种技术方案是，所述气体输入单元包括顺序连接的压力储气罐、减压阀、第一传输管、微流量计、调压阀和第二传输管，所述第二传输管分别与各集成芯片的一层结构的各通道1相连接；所述液体输入单元包括顺序连接 的储液器、第三传输管、数字控制式注射泵和第四传输管，所述第四传输管分别与各集成芯片的另一层结构的各通道1相连接。 Another technical solution of the device is that the gas input unit includes a pressure gas storage tank, a pressure reducing valve, a first delivery pipe, a micro flow meter, a pressure regulating valve and a second delivery pipe connected in sequence, and the second delivery The tubes are respectively connected to eachchannel 1 of the one-layer structure of each integrated chip; the liquid input unit includes a liquid reservoir connected in sequence, a third transfer tube, a digitally controlled syringe pump and a fourth transfer tube, the fourth The transmission pipes are respectively connected to thechannels 1 of the other layer structure of the integrated chips. the

制备微米级分散体的方法的一种技术方案是，应用于一包括集成芯片阵列的装置中，所述集成芯片阵列由集成芯片组成，其中，集成芯片为上述任一关于集成芯片的技术方案，所对应的集成芯片；装置为上述任一关于装置的技术方案，所对应的装置； A technical solution of the method for preparing a micron-scale dispersion is to apply it to a device comprising an array of integrated chips, the array of integrated chips is composed of integrated chips, wherein the integrated chip is any of the above-mentioned technical solutions about integrated chips, The corresponding integrated chip; the device is any one of the above-mentioned technical solutions about the device, and the corresponding device;

所述方法包括以下步骤：A1、按第一预设置条件，向各集成芯片的一层结构的各通道1输入第一物质；按第二预设置条件，向各集成芯片的另一层结构的各通道1输入第二物质；A2、调节第一物质和/或第二物质的输入条件，使各个第一层结构的各通道M输出的物质以同轴流动形式，由对应的各特定通道N的物质按轴向进行包裹，分别形成待收集产物流，其轴向与所述垂线重合；A3、从各收集通道的输出端收集和导出产物。 The method includes the following steps: A1. According to the first preset condition, input the first substance to eachchannel 1 of the one-layer structure of each integrated chip; Eachchannel 1 inputs the second substance; A2, adjust the input conditions of the first substance and/or the second substance, so that the substances output by each channel M of each first layer structure are coaxially flowed by the corresponding specific channels N The material is wrapped in the axial direction to form the product flow to be collected respectively, and its axial direction coincides with the vertical line; A3, collect and export the product from the output end of each collection channel. the

制备微米级分散体的方法的另一种技术方案是，所述第一物质与所述第二物质的组合，为气体与液体的组合；所述气体至少包括氮气、氟碳类气体、氟硫类气体其中之一；所述液体至少包括磷脂类液体、表面活性剂类液体、上述液体的改性液其中之一。 Another technical scheme of the method for preparing a micron-scale dispersion is that the combination of the first substance and the second substance is a combination of a gas and a liquid; the gas at least includes nitrogen, fluorocarbon gases, fluorine-sulfur One of the gas-like liquids; the liquid at least includes one of the phospholipid-based liquid, the surfactant-based liquid, and the modified liquid of the above-mentioned liquids. the

制备微米级分散体的方法的另一种技术方案是，步骤A1中，向各集成芯片的第一层结构的各通道1输入第一液体，向各集成芯片的第二层结构的各通道1输入气体或第二液体；步骤A2中，使各个第一层结构的各通道M输出的第一液体以同轴流动形式，由气体或第二液体按轴向包裹所述第一液体，分别形成待收集微液滴流。 Another technical scheme of the method for preparing micron-scale dispersions is that in step A1, the first liquid is input to eachchannel 1 of the first layer structure of each integrated chip, and the first liquid is fed to eachchannel 1 of the second layer structure of each integrated chip. Input gas or second liquid; in step A2, the first liquid output by each channel M of each first layer structure is coaxially flowed, and the first liquid is wrapped axially by gas or second liquid, respectively forming Microfluidic droplet stream to be collected. the

制备微米级分散体的方法的另一种技术方案是，步骤A1中，向各集成芯片的一层结构的各通道1输入气体，压力低于10psi，向各集成芯片的另一层结构的各通道1输入液体，流速低于3μL/s；步骤A2中，调节气体和/或液体的输入条件。 Another technical scheme of the method for preparing a micron-scale dispersion is that in step A1, the gas is input to eachchannel 1 of the one-layer structure of each integrated chip, and the pressure is lower than 10 psi, and the gas is injected into eachchannel 1 of another layer structure of each integrated chip.Channel 1 inputs liquid, and the flow rate is lower than 3 μL/s; in step A2, adjust the input conditions of gas and/or liquid. the

制备微米级分散体的方法的另一种技术方案是，步骤A1中，输入气体的压力低于5psi，输入液体的流速低于1.5μL/s。 Another technical scheme of the method for preparing a micron-scale dispersion is that in step A1, the pressure of the input gas is lower than 5 psi, and the flow rate of the input liquid is lower than 1.5 μL/s. the

制备微米级分散体的方法的另一种技术方案是，步骤A1中，所述液体预先加入特异性配体；或者，步骤A3之后，向所述产物加入特异性配体。 Another technical scheme of the method for preparing a micron-scale dispersion is that in step A1, a specific ligand is added to the liquid in advance; or, after step A3, a specific ligand is added to the product. the

制备微米级分散体的方法的另一种技术方案是，步骤A1中，向各集成芯片 的第一层结构的各通道1输入气体，向各集成芯片的第二层结构的各通道1输入液体；步骤A2中，调节气体和/或液体的输入条件，使各个第一层结构的各通道M输出的气体以同轴流动形式，按轴向包裹液体，分别形成待收集微球流。 Another technical scheme of the method for preparing the micron-scale dispersion is that, in step A1, the gas is input to eachchannel 1 of the first layer structure of each integrated chip, and the liquid is input to eachchannel 1 of the second layer structure of each integrated chip ; In step A2, adjust the input conditions of gas and/or liquid, so that the gas output from each channel M of each first layer structure flows coaxially, wraps the liquid in the axial direction, and forms the flow of microspheres to be collected respectively. the

制备微米级分散体的方法的另一种技术方案是，步骤A2中，各个第一层结构的各通道M输出的气体，分别在两侧高速流动的液体聚焦作用下形成中轴线位于所述垂线位置的稳定倒锥体，各倒锥体的尖端分别与各收集通道位置相对。 Another technical scheme of the method for preparing a micron-scale dispersion is that in step A2, the gases output by each channel M of each first layer structure are respectively formed under the focusing action of the high-speed flowing liquid on both sides, and the central axis is located at the vertical position. Stable inverted cones at the line position, the tip of each inverted cone is opposite to the position of each collection channel. the

采用上述方案，本发明提供了一种集成芯片、采用该集成芯片的装置和采用该装置制备微米级分散体的方法。在此基础上，本发明提供了一种实用的大规模微流控流动聚焦装置，以及粒径均一、可控制、单分散的声学性质稳定的超声造影剂制备方法，以制备粒径大小可控并且呈单分散性的超声造影剂；本超声造影剂制备简便、效果好且安全，制备的微泡外壳厚度均匀，具有很高的应用价值。 Using the above solution, the present invention provides an integrated chip, a device using the integrated chip and a method for preparing micron-scale dispersions using the device. On this basis, the present invention provides a practical large-scale microfluidic flow focusing device, and a method for preparing an ultrasonic contrast agent with uniform particle size, controllability, and monodisperse acoustic properties, so as to prepare a controllable particle size And it is a monodisperse ultrasonic contrast agent; the ultrasonic contrast agent is easy to prepare, has good effect and is safe, and the prepared microbubble shell has uniform thickness, and has high application value. the

最显著的优点是制作的超声造影剂微泡具有高度单分散性和粒径可控性，满足超声造影成像技术的要求。微泡粒径的多分散性指数＜2％，粒径随着气体压力的增加而变大，随着液体流速的增加而减小，控制非常灵活；同时，采用多个收集通道所形成的微喷嘴阵列大大提高了微泡的制备效率；生产装置具有重复使用性，降低了生产成本。 The most notable advantage is that the produced ultrasound contrast agent microbubbles have high monodispersity and particle size controllability, which meets the requirements of ultrasound contrast imaging technology. The polydispersity index of the microbubble particle size is less than 2%. The particle size becomes larger with the increase of gas pressure and decreases with the increase of liquid flow rate. The control is very flexible; at the same time, the microbubble formed by multiple collection channels The nozzle array greatly improves the preparation efficiency of the microbubbles; the production device is reusable, which reduces the production cost. the

并且，本发明的装置和方法可用于制备多种类型的微米级分散体，包括超声造影剂微泡，以及微液滴或其他微型混和物。常规的超声造影剂可直接用该装置制备，如脂类、白蛋白类、多聚体类、表面活性剂类等；通过加入特异性配体，还可以制备靶向超声造影剂，有些壳材，如蛋白分子，在高温和超声条件下失活，所以本发明方法解决了以往声振法所不能的解决的技术问题。该发明装置对这两种方法都适用，特别是当采用后一种选择时更具有优势，其优点一是减少了制备环节，二是减少了制备环节微泡的破坏。而且，本发明装置在造影剂制备过程中产热少，因此尤其适合用于制备兼药物或基因靶向载体的超声造影剂。 Moreover, the device and method of the present invention can be used to prepare various types of micron-scale dispersions, including ultrasound contrast agent microbubbles, and micro-droplets or other micro-mixtures. Conventional ultrasound contrast agents can be prepared directly with this device, such as lipids, albumins, polymers, surfactants, etc.; by adding specific ligands, targeted ultrasound contrast agents can also be prepared, and some shell materials , such as protein molecules, are inactivated under high temperature and ultrasonic conditions, so the method of the present invention solves the technical problems that cannot be solved by the acoustic vibration method in the past. The device of the invention is applicable to both methods, especially when the latter option is used, it has more advantages. The first advantage is that the preparation process is reduced, and the second is that the destruction of microbubbles in the preparation process is reduced. Moreover, the device of the present invention generates less heat during the preparation of the contrast agent, so it is especially suitable for the preparation of an ultrasound contrast agent that is also a drug or gene targeting carrier. the

【附图说明】【Description of drawings】

图1为本发明实施例的超声造影剂制备装置的示意图； Fig. 1 is the schematic diagram of the ultrasonic contrast medium preparation device of the embodiment of the present invention;

图2是本发明实施例的微流控大规模集成芯片的底层通道结构示意图； Fig. 2 is a schematic diagram of the underlying channel structure of the microfluidic large-scale integrated chip of the embodiment of the present invention;

图3是本发明实施例的微流控大规模集成芯片的顶层通道结构示意图； 3 is a schematic diagram of the top-level channel structure of the microfluidic large-scale integrated chip of the embodiment of the present invention;

图4是图2虚线框所示的喷嘴结构放大示意图； Figure 4 is an enlarged schematic view of the nozzle structure shown in the dotted line box in Figure 2;

图5是本发明实施例的微泡形成示意图； Fig. 5 is the microbubble formation schematic diagram of the embodiment of the present invention;

图6是本发明另一实施例的微流控大规模集成芯片的示意图； Fig. 6 is the schematic diagram of the microfluidic large-scale integrated chip of another embodiment of the present invention;

图7是图6虚线框所示结构的放大图； Figure 7 is an enlarged view of the structure shown in Figure 6 dotted line box;

图8是本发明实验一生成的微泡照片； Fig. 8 is the microbubble photograph that experiment one of the present invention generates;

图9是本发明实施例装置的另一种喷嘴结构示意图； Fig. 9 is another kind of nozzle structure schematic diagram of the embodiment device of the present invention;

图10是本发明实施例的连通腔中，各特定通道N的出口所在平面的示意图； Fig. 10 is a schematic diagram of the plane where the outlet of each specific channel N is located in the communicating cavity of the embodiment of the present invention;

图11是图10所示的纵向平面的示意图。 FIG. 11 is a schematic view of the longitudinal plane shown in FIG. 10 . the

【具体实施方式】【Detailed ways】

以下结合附图和具体实施例进行详细说明。 A detailed description will be given below in conjunction with the accompanying drawings and specific embodiments. the

超声造影剂微泡的粒径大小及其分布是超声造影剂的重要指标，然而，目前的超声造影剂制作方法如声振法、机械振荡法产生的微泡粒径分布范围较宽，且主要通过手摇或者振荡的方式形成，声学性质不稳定。因此，提出一种大规模微流控超声造影剂的制备方法及装置，结合目前较成熟的微纳米加工技术制作微流控大规模集成芯片，利用流动聚焦原理，制备出粒径大小可控制且高度单分散性的微泡，微泡外壳厚度均匀，不仅可以获得稳定的声学性质，也可以减少不同大小微泡之间的融合成大微泡的可能，具有更高的有效性和安全性，操作具有较好的重复性和可控性。 The particle size and distribution of the microbubbles of ultrasound contrast agents are important indicators of ultrasound contrast agents. Formed by shaking or oscillating, the acoustic properties are unstable. Therefore, a method and device for preparing a large-scale microfluidic ultrasound contrast agent is proposed, combined with the current relatively mature micro-nano processing technology to produce a microfluidic large-scale integrated chip, and using the principle of flow focusing to prepare a controllable particle size and Highly monodisperse microbubbles, uniform thickness of the microbubble shell, not only can obtain stable acoustic properties, but also reduce the possibility of fusion between different sizes of microbubbles into large microbubbles, with higher effectiveness and safety, The operation has good repeatability and controllability. the

因此，本发明提出一种采用了微流控技术的集成芯片，通过微流控方式，可大规模地制备微米级分散体，包括超声造影剂或其他微型混和物。 Therefore, the present invention proposes an integrated chip using microfluidic technology, through which micron-scale dispersions, including ultrasound contrast agents or other micro-mixtures, can be prepared on a large scale. the

集成芯片的基质为表面亲水性基质，所述表面亲水性基质包括亲水性基质、以及表面进行过亲水性处理的疏水性基质；需要说明的是，通道的材料不一定要有亲水性，但是，疏水性材料的效果没有亲水性材料的效果好，对于疏水性的材料，可将其表面处理为亲水性，也能获得类似于亲水性材料的效果；表面为亲水性的基质，可以很顺畅地产生微泡。 The substrate of the integrated chip is a surface hydrophilic substrate, and the surface hydrophilic substrate includes a hydrophilic substrate and a hydrophobic substrate with a hydrophilic treatment on the surface; it should be noted that the material of the channel does not necessarily have to be hydrophilic. Water-based, however, the effect of hydrophobic materials is not as good as that of hydrophilic materials. For hydrophobic materials, the surface can be treated to be hydrophilic, and the effect similar to hydrophilic materials can also be obtained; the surface is hydrophilic The water-based base can generate microbubbles smoothly. the

一个例子是，其基质为硅、玻璃、聚二甲基硅氧烷、聚甲基丙烯酸甲酯或聚碳酯。其中，工艺上可满足结构要求、不易与通道内的气体、液体发生反应的材料均可作为基质，本发明对此不作任何额外限制；例如，聚二甲基硅氧烷 具有良好的绝缘性，能承受高电压；热稳定性高，适合加工各种反应芯片；具有优良的光学特性，可应用于多种光学检测系统；此外，它还可以和硅、氮化硅、氧化硅、玻璃等许多材料形成很好的密封，其还具有高疏水性以及对生物大分子较强的表面吸附特性，因此已广泛应用于微流控芯片研究领域。 An example is where the substrate is silicon, glass, polydimethylsiloxane, polymethylmethacrylate or polycarbonate. Among them, the material that can meet the structural requirements in the process and is not easy to react with the gas and liquid in the channel can be used as the substrate, and the present invention does not make any additional restrictions on this; for example, polydimethylsiloxane has good insulation, Can withstand high voltage; high thermal stability, suitable for processing various reaction chips; has excellent optical properties, can be applied to a variety of optical detection systems; in addition, it can also be used with silicon, silicon nitride, silicon oxide, glass and many other The material forms a good seal, and it also has high hydrophobicity and strong surface adsorption characteristics for biomacromolecules, so it has been widely used in the field of microfluidic chip research. the

以两层相邻结构为一组结构，所述集成芯片包括至少一组结构；即所述集成芯片可以包括两层结构，或者包括四层结构、六层结构、八层结构甚至更多。例如，所述集成芯片仅设置一组结构。即，所述集成芯片为偶数层结构，进行上下堆叠或倾斜堆叠而形成，各层结构的具体特征如下所述：所述集成芯片任一端的边缘层结构，与该边缘层结构相邻的层结构，组成了一组结构；所述集成芯片可以有一组结构，也可以有多组结构。集成芯片也可以平行排列而形成，即一组结构或多组上下堆叠的多层结构平行排列。 Taking two adjacent structures as a group structure, the integrated chip includes at least one group of structures; that is, the integrated chip may include a two-layer structure, or include a four-layer structure, six-layer structure, eight-layer structure or even more. For example, the integrated chip only has one set of structures. That is, the integrated chip has an even-numbered layer structure, which is formed by stacking up and down or obliquely. The specific features of each layer structure are as follows: the edge layer structure at either end of the integrated chip, the layer adjacent to the edge layer structure Structures constitute a group of structures; the integrated chip may have a group of structures, or may have multiple groups of structures. Integrated chips can also be formed in parallel arrangement, that is, one set of structures or multiple sets of stacked multilayer structures are arranged in parallel. the

在一组结构中，具有两层相邻的结构，其中，第一层结构设置M级梯度，第二层结构设置N级梯度；其中，M、N是自然数，M小于N；各级梯度中，第一级梯度设置H1个通道1，第二级梯度从每个通道1中引出H2个分支，形成H1×H2个通道2，以此类推，第N级梯度从每个通道(N-1)中引出HN个分支，形成H1×H2......×HN个通道N。优选的例子是，每级分支相对于前一级分叉的分支要均匀分布，主要是为了保证最后各通道出来的同样的液体的流速一致，或同样的气体的流速一致；即，第N级梯度从每个通道(N-1)中引出HN个分支，某一通道(N-1)所引出的HN个分支，相对于该通道均匀分布。 In a set of structures, there are two adjacent structures, wherein the first layer structure sets M-level gradients, and the second layer structure sets N-level gradients; where M and N are natural numbers, and M is less than N; , the first-level gradient setsH1 channels 1, the second-level gradient leads to H2 branches from eachchannel 1, forming H1×H2 channels 2, and so on, the Nth-level gradient starts from each channel (N-1 ) leads to HN branches to form H1×H2...×HN channels N. A preferred example is that the branches of each stage are evenly distributed with respect to the branches of the previous stage, mainly to ensure that the flow velocity of the same liquid coming out of each channel at the end is consistent, or the flow velocity of the same gas is consistent; that is, the Nth stage The gradient derives HN branches from each channel (N-1), and the HN branches derived from a certain channel (N-1) are evenly distributed with respect to the channel. the

其中，第一层结构的各通道1，用于输入第一种物质；第二层结构的各通道1，用于输入第二种物质。第一种物质和第二种物质，可以为不同的气体或液体，也可以为两种不同的液体，还可以是，第一种物质为气体，第二种物质为液体，也可以是，第一种物质为液体，第二种物质为气体。例如，第一层结构作为气体通道，第二层结构作为液体通道；或者，第一层结构作为液体通道，第二层结构作为气体通道；或者，第一层结构作为第一种液体通道，第二层结构作为第二种液体通道。 Wherein, eachchannel 1 of the first layer structure is used to input the first substance; eachchannel 1 of the second layer structure is used to input the second substance. The first substance and the second substance may be different gases or liquids, or two different liquids. It may also be that the first substance is a gas and the second substance is a liquid, or that the first substance is a liquid. One substance is a liquid and the second substance is a gas. For example, the first layer structure is used as a gas channel, and the second layer structure is used as a liquid channel; or, the first layer structure is used as a liquid channel, and the second layer structure is used as a gas channel; or, the first layer structure is used as the first liquid channel, and the second layer structure is used as a liquid channel. The two-layer structure acts as a second liquid channel. the

例如，第一层结构作为气体通道，第二层结构作为液体通道；此时，第一层结构的H1个通道1分别连接气体输入口，第二层结构的H1个通道1分别连接液体输入口；第一层结构的H1×H2......×HM个通道M分别连接气体输出口，第二层结构的H1×H2......×HN个通道N连接液体输出口。或者，第一层 结构作为液体通道，第二层结构作为气体通道，其余类似，不做赘述。 For example, the first layer structure is used as a gas channel, and the second layer structure is used as a liquid channel; at this time, theH1 channels 1 of the first layer structure are respectively connected to the gas input port, and theH1 channels 1 of the second layer structure are respectively connected to the liquid input port ; The H1×H2...×HM channels M of the first layer structure are respectively connected to the gas outlet, and the H1×H2...×HN channels N of the second layer structure are connected to the liquid output port. Alternatively, the first layer structure is used as a liquid channel, and the second layer structure is used as a gas channel, and the rest are similar and will not be described in detail. the

一个优选的例子是，M＝N-1，即M比N小1，第一层结构的梯度数比第二层结构的梯度数少一层。例如，N为8，M为7。 A preferred example is, M=N-1, that is, M is 1 smaller than N, and the gradient number of the first layer structure is one layer less than that of the second layer structure. For example, N is 8 and M is 7. the

或者，M比N小2、3、4、......等等，例如，N为8，M为4；又如，N为6，M为3；又如，N为9，M为7；又如，N为21，M为12。本发明对此不做额外限制，只需能够实现第一层结构与第二层结构梯度数的差异即可。 Or, M is smaller than N by 2, 3, 4, ... etc., for example, N is 8, M is 4; another example, N is 6, M is 3; another example, N is 9, M is 7; for another example, N is 21, and M is 12. The present invention does not impose additional restrictions on this, and only needs to be able to realize the difference in gradient numbers between the first layer structure and the second layer structure. the

其中，H1、H2、H3......HM、H(M+1)、H(M+2)......H(N-2)、H(N-1)、HN可以全部相同，也可以部分相同，还可以均不相同。例如，H1、H2、H3......HM、H(M+1)、H(M+2)......HN均为2，并且，M＝N-1，这样通道N的数量是通道M的2倍；这种情况下，由于H1为2，因此有两个气体入口和两个液体入口；或者，H1为2，H2至HN均为3；或者，H1为1，H2为2、H3为3、H4为4......HN为N；或者，其均不相同，如H1为4，H2为1、H3为6、H4为5......HN为2。本发明对此不做额外限制，只需能够实现各级分支结构即可。 Among them, H1, H2, H3...HM, H(M+1), H(M+2)...H(N-2), H(N-1), HN can All are the same, some may be the same, or all may be different. For example, H1, H2, H3...HM, H(M+1), H(M+2)...HN are all 2, and M=N-1, so channel N The number of channels is twice that of M; in this case, since H1 is 2, there are two gas inlets and two liquid inlets; or, H1 is 2, and H2 to HN are 3; or, H1 is 1, H2 is 2, H3 is 3, H4 is 4...HN is N; or, they are all different, such as H1 is 4, H2 is 1, H3 is 6, H4 is 5... HN is 2. The present invention does not impose additional restrictions on this, as long as the branch structure at all levels can be realized. the

对于通道的形状，各通道的横截面优选为矩形；其中，通道M横截面与通道N横截面，可以为形状相同或者不相同的矩形，例如，所述矩形的高度为20μm至30μm，通道M横截面的宽度为30μm至50μm，通道N横截面的宽度为50μm至100μm；或者，通道M横截面的宽度为50μm至100μm，通道N横截面的宽度为30μm至50μm；或者，通道M横截面与通道N横截面的宽度均为30μm至50μm。 Regarding the shape of the channel, the cross section of each channel is preferably rectangular; wherein, the cross section of channel M and the cross section of channel N can be rectangles with the same or different shapes, for example, the height of the rectangle is 20 μm to 30 μm, channel M The width of the cross section is 30 μm to 50 μm, and the width of the channel N cross section is 50 μm to 100 μm; or, the width of the channel M cross section is 50 μm to 100 μm, and the width of the channel N cross section is 30 μm to 50 μm; alternatively, the width of the channel M cross section The width of the cross-section of the channel N is 30 μm to 50 μm. the

一个例子是，通道为均一或不均一的长方体，但是截面一定是矩形，例如通道为一个正四棱锥台、一个横截面为相似矩形的四棱锥台、或一个梯形台。一个优选的例子是，各通道为长方体，通道的横截面为矩形，其高度为20μm至30μm，宽度为30μm至100μm。例如，该矩形的高度为25μm，宽度为40μm；又如，该矩形的高度为28μm，宽度为35μm；又如，该矩形的高度为22μm，宽度为45μm；又如，该矩形的高度为21μm，宽度为85μm；又如，该矩形的高度为29μm，宽度为65μm；又如，该矩形的高度为29μm，宽度为75μm。 An example is that the channel is a uniform or non-uniform cuboid, but the cross-section must be rectangular, for example, the channel is a square pyramid, a square pyramid with a similar rectangular cross section, or a trapezoidal platform. A preferred example is that each channel is a cuboid, the cross section of the channel is rectangular, its height is 20 μm to 30 μm, and its width is 30 μm to 100 μm. For example, the height of the rectangle is 25 μm and the width is 40 μm; as another example, the height of the rectangle is 28 μm and the width is 35 μm; as another example, the height of the rectangle is 22 μm and the width is 45 μm; as another example, the height of the rectangle is 21 μm , the width is 85 μm; as another example, the height of the rectangle is 29 μm, and the width is 65 μm; as another example, the height of the rectangle is 29 μm, and the width is 75 μm. the

一个优选的例子是，通道M横截面与通道N横截面，为高度相同的矩形。 A preferred example is that the cross section of channel M and the cross section of channel N are rectangles with the same height. the

也就是说，对于某一通道，垂直于其通道方向的截面，即该通道的通道截面作为所述横截面，为正方形或长方形，需要说明的是，通道截面还可以为三角形、菱形、五边形或其他形状，本发明对此不做额外限制；对于各通道而言， 只需其通道截面的面积，至少为600平方微米即可，例如，其通道截面的面积为800、900或1000平方微米。又如，各通道为圆柱体或其他形体；或者，各通道还可以是弯折变向的方形体、圆形体或螺旋状，例如呈L形、Y形等；此时，通道的输出方向为其最后一段未弯折通道的方向，例如，L形通道的输出方向为变向后具有出口的一段通道的方向。 That is to say, for a channel, the section perpendicular to its channel direction, that is, the channel section of the channel as the cross section, is a square or a rectangle. It should be noted that the channel section can also be triangular, rhombus, five-sided Shape or other shapes, the present invention does not make additional restrictions on this; for each channel, it only needs the area of its channel cross-section to be at least 600 square microns, for example, the area of its channel cross-section is 800, 900 or 1000 square microns Micron. As another example, each channel is a cylinder or other shapes; or, each channel can also be a square body, a circular body or a spiral shape that bends and changes direction, such as L-shaped, Y-shaped, etc.; at this time, the output direction of the channel It is the direction of the last unbent channel, for example, the output direction of the L-shaped channel is the direction of the channel with the outlet after the change of direction. the

在一组结构中，对于第一层结构的任一特定通道M，均对应于第二层结构从一通道M所引出的HM个分支中的某一分支；该分支在第M+1级梯度形成H(M+1)个分支......在第N级梯度形成H(M+1)×H(M+2)......×H(N-1)×HN数量的特定通道N；即对于第一层结构的任一通道M，在第二层结构中，与该第一层结构通道M对应的第二层结构通道M在下一级梯度将引出H(M+1)个分支，这些分支最终在第N级梯度形成H(M+1)×H(M+2)......×H(N-1)×HN个通道N，与该第一层结构通道M相对应。 In a group of structures, for any specific channel M of the first layer structure, it corresponds to a certain branch of the HM branches derived from a channel M of the second layer structure; the branch at the M+1 level gradient Form H(M+1) branches... Form H(M+1)×H(M+2) at N level gradient...×H(N-1)×HN number The specific channel N of ; that is, for any channel M of the first layer structure, in the second layer structure, the channel M of the second layer structure corresponding to the channel M of the first layer structure will lead to H(M+ 1) branches, these branches finally form H(M+1)×H(M+2)…×H(N-1)×HN channels N at the N-level gradient, and the first Layer structure channel M corresponds. the

其中，各特定通道N的出口等距分布在一预设置半径圆形的圆周，即，这些通道N的出口排列形成正多边形，并位于正多边形上的各个顶点；例如，这些通道N的出口形成正三角形、正方形、正五边形、正六边形......正99边形、正100边形、正101边形等等；特殊的，当仅有两个通道N时，出口位于该预设置半径圆形的某一直径的两端。其中，预设置半径可根据实际情况而设置，例如，根据不同的反应物、不同的反应条件，预设置该圆形半径的大小。圆形的半径与通道的尺寸有关系，以第一层通道输入气体，第二层通道输入液体为例，气体出口垂直于前述圆形时，液体出口均布在气体出口的周边即可。例如，该圆形半径为150μm至1500μm，更具体的例子是，该圆形半径可为250μm、500μm、700μm、1100μm、1200μm、1400μm等等。优选的是，该圆形半径与通道的宽度相对应，通道的宽度越大，则该圆形半径越大。 Wherein, the outlets of each specific channel N are equidistantly distributed on the circumference of a preset radius circle, that is, the outlets of these channels N are arranged to form a regular polygon, and are located at each vertex on the regular polygon; for example, the outlets of these channels N form Regular triangle, square, regular pentagon, regular hexagon...regular 99-gon, regular 100-gon, regular 101-gon, etc.; special, when there are only two channels N, the outlet is located Both ends of a certain diameter of the preset radius circle. Wherein, the preset radius can be set according to the actual situation, for example, the size of the circular radius can be preset according to different reactants and different reaction conditions. The radius of the circle is related to the size of the channel. Taking the first layer of channel to input gas and the second layer of channel to input liquid as an example, when the gas outlet is perpendicular to the aforementioned circle, the liquid outlet can be evenly distributed around the gas outlet. For example, the radius of the circle is 150 μm to 1500 μm, more specifically, the radius of the circle can be 250 μm, 500 μm, 700 μm, 1100 μm, 1200 μm, 1400 μm and so on. Preferably, the radius of the circle corresponds to the width of the channel, the greater the width of the channel, the greater the radius of the circle. the

需要说明的是，当尺度较小时，例如，以微米级进行量度时，通道的出口、以及收集通道的输入端，往往是一个面的概念；而本发明各实施例中，凡是涉及到点与点之间的连线，应理解为出口面的中心点，或者，输入端的面的中心点；例如，一个三角形或矩形的几何中心点；或者，大致为出口面的中心点，或输入端的面的中心点。这样，出口排列形成正多边形，才能获得更好的精度。并且，出口之间的连线、出口与收集通道的输入端之间的连线才能更精确，以获得更好的效果。 It should be noted that when the scale is small, for example, when measuring at the micron level, the outlet of the channel and the input end of the collection channel are often a concept of a surface; A line between points is to be understood as the center point of the outlet face, or, the center point of the input face; for example, the geometric center point of a triangle or rectangle; or, approximately the center point of the outlet face, or the input face the center point of . In this way, the outlet arrangement forms a regular polygon to obtain better precision. In addition, the connection between the outlets and the connection between the outlet and the input end of the collection channel can be more precise, so as to obtain better effects. the

一个例子如图10所示，有六个特定通道N，其出口等距分布在一预设置半径圆形的圆周，六个出口形成一正六边形，位于一横向平面中，该横向平面位于一连通腔中，液体分别从各个特定通道N的出口流向该预设置半径圆形的圆心，即，液体出口到圆心的距离相同，并且排列为正六边形；并且，正六边形中，任意两个相对的特定通道N，位于一个纵向平面内；例如任意两个相对的特定通道N，与该预设置半径圆形的圆心形成对称或大致对称关系。 An example is shown in Figure 10, there are six specific channels N, the outlets of which are equidistantly distributed on the circumference of a circle with a preset radius, and the six outlets form a regular hexagon and are located in a transverse plane, which is located in a In the communication cavity, the liquid flows from the outlet of each specific channel N to the center of the preset radius circle, that is, the distance from the liquid outlet to the center of the circle is the same, and the arrangement is a regular hexagon; and, in the regular hexagon, any two The relative specific passages N are located in a longitudinal plane; for example, any two relative specific passages N form a symmetrical or approximately symmetrical relationship with the center of the circle with a preset radius. the

上例中，任一纵向平面如图11所示，两个相对的特定通道N，对于该特定通道M的出口与该圆形的垂线，形成轴对称关系；第一层气体从出口输出之后，继续再流动一段距离，即一段特定长度，该特定长度主要是与第二层液体通道矩形横截面的高度或宽度有关系，例如，该特定长度为液体通道横截面的宽度或高度的25％至75％，如通道横截面的高度的30％、40％、50％、60％、70％等等，优选的例子是，当只有两个液体通道和一个气体通道时，所有出口全部在一个平面内，如图4和图5所示，气体通道出口与圆心的距离就是液体通道横截面的宽度的一半；当有多个液体通道，气体通道出口垂直于液体通道出口所在平面时，气体通道出口与圆心的距离就是液体通道横截面的高度的一半。之后气体与液体混合，在进入所述收集通道的输入端之前，形成待收集产物流，通过收集通道进行收集；例如，形成倒锥体或其他形状的待收集产物流，通过收集通道进行收集。 In the above example, as shown in Figure 11 on any longitudinal plane, two relative specific channels N form an axisymmetric relationship between the outlet of the specific channel M and the vertical line of the circle; after the first layer of gas is output from the outlet , continue to flow for a certain distance, that is, a specific length, which is mainly related to the height or width of the rectangular cross-section of the second-layer liquid channel, for example, the specific length is 25% of the width or height of the liquid channel cross-section to 75%, such as 30%, 40%, 50%, 60%, 70% of the height of the channel cross-section, etc., a preferred example is that when there are only two liquid channels and one gas channel, all outlets are in one In the plane, as shown in Figure 4 and Figure 5, the distance between the outlet of the gas channel and the center of the circle is half the width of the cross-section of the liquid channel; The distance between the outlet and the center of the circle is half the height of the liquid passage cross section. The gas is then mixed with the liquid to form a stream of product to be collected, which is collected through the collection channel, before entering the input end of said collection channel; for example, an inverted cone or other shaped stream of product to be collected, which is collected through the collection channel. the

上面这两个例子，描述的是偶数个特定通道N，任一纵向平面具有形成轴对称关系或大致对称的一对特定通道N；需要说明的是，如果是奇数个特定通道N，虽然在纵向平面上不能形成一对特定通道N，但是，同样可实现本发明的效果。例如，五个特定通道N，其出口等距分布在一预设置半径圆形的圆周，五个出口形成一正五边形，位于一横向平面中，该横向平面位于一连通腔中，液体分别从各个特定通道N的出口流向该预设置半径圆形的圆心。 The above two examples describe an even number of specific channels N, and any longitudinal plane has a pair of specific channels N that form an axisymmetric relationship or are roughly symmetrical; it should be noted that if there are an odd number of specific channels N, although in the longitudinal direction A pair of specific channels N cannot be formed on a plane, but the effect of the present invention can also be achieved. For example, five specific channels N, the outlets of which are equidistantly distributed on the circumference of a circle with a preset radius, and the five outlets form a regular pentagon, are located in a transverse plane, and the transverse plane is located in a communicating chamber, and the liquids are respectively Flow from the outlet of each specific channel N to the center of the circle with a preset radius. the

该特定通道M的输出方向垂直于该圆形，该特定通道M的出口与该圆形的垂线通过其圆心，即这个通道M的输出方向与圆形所在的平面相垂直，这个通道M的出口在该平面的投影位于该圆形的圆心。各特定通道N的输出方向与所述垂线的夹角相同，并且，这些夹角为锐角；例如，各特定通道N的输出方向通过该圆形圆心，各特定通道N根据其数量，位于该圆形的各等分线上，假设有3个特定通道N，则相邻两个特定通道N之间的夹角为120度；或者，各特 定通道N可组成一个直圆锥体，该直圆锥体以该特定通道M的输出方向为旋转轴，各特定通道N与该旋转轴的夹角相同，并且，这些夹角为锐角；例如，夹角均为30度、45度、60度、75度或80度等等。例如，由于各特定通道N的出口等距分布在一预设置半径圆形的圆周，当各特定通道N的数量为偶数时，对称于该旋转轴的两条特定通道N的出口，所对应的两条特定通道N形成一包括该旋转轴的平面，并且，两条特定通道N在其输出方向上，可形成一等腰梯形。例如，各特定通道N形成一种对称的形状；例如为一个半截的六棱锥体，即一个六棱锥台，其底面为正六边形，六棱可组成三个等腰梯形。 The output direction of the specific channel M is perpendicular to the circle, and the vertical line between the outlet of the specific channel M and the circle passes through the center of the circle, that is, the output direction of the channel M is perpendicular to the plane where the circle is located, and the output direction of the channel M The projection of the outlet on the plane is located at the center of the circle. The output direction of each specific channel N has the same angle with the vertical line, and these included angles are acute angles; for example, the output direction of each specific channel N passes through the center of the circle, and each specific channel N is located at the On each bisector of the circle, assuming that there are 3 specific channels N, the angle between two adjacent specific channels N is 120 degrees; or, each specific channel N can form a straight cone, and the straight The cone takes the output direction of the specific channel M as the axis of rotation, and the included angles between each specific channel N and the axis of rotation are the same, and these included angles are acute angles; for example, the included angles are 30 degrees, 45 degrees, 60 degrees, 75 degrees or 80 degrees and so on. For example, since the outlets of each specific channel N are equidistantly distributed on the circumference of a preset radius circle, when the number of each specific channel N is an even number, the outlets of two specific channels N symmetrical to the rotation axis, the corresponding The two specific channels N form a plane including the rotation axis, and the two specific channels N can form an isosceles trapezoid in their output direction. For example, each specific channel N forms a symmetrical shape; for example, it is a half-truncated hexagonal pyramid, that is, a hexagonal truncated pyramid, the base of which is a regular hexagon, and the six edges can form three isosceles trapezoids. the

这样，在一组结构中，通过调节一层结构的输入条件与另一层结构的输入条件，使各个第一层结构的各通道M输出的物质以同轴流动形式，由对应的各特定通道N的物质按轴向包裹某一通道M输出的物质，即以某一通道M输出的物质的流向为轴，通过输出对应的各特定通道N的物质，从而形成一层层的覆以对应的各特定通道N输出的物质；或者说，各特定通道N输出的物质沿轴向，以同心圆形式覆盖在通道M输出的物质上；分别形成待收集产物流，例如，待收集产物流形成一倒锥体；其轴向与所述旋转轴重合。即某个第一层结构的某一通道M输出的物质，一直沿着其输出方向流动，或者以螺旋形绕着其输出方向流动；以其输出方向为轴向，由其对应的各特定通道N的物质按轴向包裹之。 In this way, in a group of structures, by adjusting the input conditions of one layer of structure and the input conditions of another layer of structure, the substances output by each channel M of each first layer of structure are coaxially flowed by the corresponding specific channels The substance of N wraps the substance output by a certain channel M in the axial direction, that is, the flow direction of the substance output by a certain channel M is taken as the axis, and the substances of the corresponding specific channels N are output to form a layer of material covered with the corresponding The material output by each specific channel N; or in other words, the material output by each specific channel N covers the material output by the channel M in the form of concentric circles along the axial direction; respectively form the product flow to be collected, for example, the product flow to be collected forms a Inverted cone; its axis coincides with the axis of rotation. That is, the substances output by a certain channel M of a certain first-layer structure always flow along its output direction, or flow around its output direction in a spiral shape; with its output direction as the axial direction, each specific channel corresponding to it The substance of N wraps it axially. the

各特定通道N的出口与该特定通道M的出口连通设置；例如，各特定通道N的出口通过一连通腔与该特定通道M的出口连通设置；本发明各例中，对连通腔不作任何限制，只需能够实现形成一待收集产物流并通过收集槽输出即可，也可以不设置该连通腔。 The outlet of each specific channel N communicates with the outlet of the specific channel M; for example, the outlet of each specific channel N communicates with the outlet of the specific channel M through a communication cavity; in the examples of the present invention, no restrictions are made to the communication cavity , it only needs to be able to realize the formation of a product flow to be collected and output through the collection tank, and the communication chamber may not be provided. the

沿该特定通道M的输出方向，在该圆形远离该特定通道M的一侧，对应设置一收集通道；用于在所述收集通道的输入端之前，形成待收集产物流，在收集通道的输入端收集产物，通过收集通道输出到收集槽；即设置了与通道M的数量相同的收集通道，对于各个特定通道M，分别沿其输出方向设置一收集通道，特定通道M与其收集通道的输出方向相同，同在一直线上。 Along the output direction of the specific channel M, on the side of the circle away from the specific channel M, a collection channel is correspondingly arranged; it is used to form a flow of products to be collected before the input end of the collection channel, and on the side of the collection channel The input end collects the product, and outputs it to the collection tank through the collection channel; that is, the same number of collection channels as the number of channels M is set, and for each specific channel M, a collection channel is set along its output direction, and the output of the specific channel M and its collection channel In the same direction, on a straight line. the

一个例子是，所述收集通道的结构包括以下结构的一种或多种：外向放大的锥体、球体、半球体或锥台体，锥台体即一个锥体切去尖端的剩余台体部分，其输入端的管径为7μm至25μm，高度为20μm至30μm；或者，圆柱体或长方 体，如图9所示，其沿对应的特定通道M输出方向的通道截面的面积略小于该特定通道M沿该方向的通道截面的面积，高度为20μm至30μm，此时，需要控制圆柱体或长方体通道的长度，使得微泡正好在收集槽中产生，以具备速度梯度，从而产生剪切力。例如，收集通道的结构为外向放大的锥体，或者，收集通道的结构为外向放大的锥台体与圆柱体的组合。 An example is that the structure of the collection channel includes one or more of the following structures: outwardly enlarged cones, spheres, hemispheres or frustums, that is, a cone cuts off the remaining frustum portion of the tip , the diameter of the input end is 7 μm to 25 μm, and the height is 20 μm to 30 μm; or, a cylinder or a cuboid, as shown in Figure 9, the area of the channel section along the output direction of the corresponding specific channel M is slightly smaller than the specific channel M The area of the channel section of the channel M along this direction has a height of 20 μm to 30 μm. At this time, the length of the cylindrical or cuboid channel needs to be controlled so that the microbubbles are just generated in the collection tank to have a velocity gradient and generate shear force . For example, the structure of the collection channel is an outwardly enlarged cone, or the structure of the collection channel is a combination of an outwardly enlarged frustum and a cylinder. the

所述集成芯片还包括至少一与各收集通道的输出端相连接的收集槽，用于收集和导出产物。 The integrated chip also includes at least one collection tank connected to the output end of each collection channel for collecting and discharging products. the

并且，利用现有的微纳米级加工技术，经过甩胶、曝光、显影、浇注、剥离、粘合等步骤，可以制作出基于聚二甲基硅氧烷(Polydimethylsiloxane，PDMS)、聚甲基丙烯酸甲酯(PMMA)或聚碳酯(PC)的微流控大规模集成芯片。 Moreover, using the existing micro-nano-scale processing technology, through steps such as glue rejection, exposure, development, pouring, peeling, and bonding, it is possible to produce polydimethylsiloxane (Polydimethylsiloxane, PDMS), polymethacrylic acid-based Methyl ester (PMMA) or polycarbonate (PC) microfluidic large-scale integration chip. the

例如，H1为1，H2至HN均为2，如图11所示，所述特定通道M的出口与其对应收集通道输入端的连线，垂直于两个特定通道N出口的连线，即每个特定通道M对应于两个特定通道N，两个特定通道N各自的出口，两者连成线段的中线为特定通道M的出口与对应的收集通道的输入端的连线；并且，通常情况下，这两个特定通道N关于上述中线对称设置；也就是说，这是一个特例，此时，两个出口不能形成一个确定的圆形，仍是等距分布在一预设置半径圆形的圆周，因此，两个特定通道N各自的出口，两者连成线段的中垂线为特定通道M的出口与对应的收集通道的输入端的连线。由于两个特定通道N出口所构成的圆形不是唯一的，因此前述连线亦是属于特定通道M的出口与所述圆形圆心的垂线之一。 For example, H1 is 1, and H2 to HN are all 2. As shown in Figure 11, the connection between the outlet of the specific channel M and its corresponding collection channel input is perpendicular to the connection between the two specific channel N outlets, that is, each The specific channel M corresponds to two specific channels N, the respective outlets of the two specific channels N, and the midline connecting the two is the connection between the outlet of the specific channel M and the input end of the corresponding collection channel; and, usually, The two specific channels N are arranged symmetrically with respect to the above-mentioned midline; that is, this is a special case, at this time, the two outlets cannot form a definite circle, but are still equidistantly distributed on the circumference of a circle with a preset radius, Therefore, the respective outlets of the two specific channels N, the perpendicular line connecting the two is the connection between the outlet of the specific channel M and the input end of the corresponding collection channel. Since the circle formed by the outlets of the two specific channels N is not unique, the aforementioned connecting line is also one of the perpendicular lines between the outlets of the specific channel M and the center of the circle. the

这样，当N为9，M为8时，则第一层结构最终形成2的(M-1)次方，即2的7次方，即128个通道出口，第二层结构最终形成2的(N-1)次方，即2的8次方，即256个通道出口；也就是说，其具有128个通道M，每个通道M对应着两个通道N，共有128个输出端；例如通道M1对应着通道N11和通道N12，通道N11和通道N12的输出方向相对，并位于同一直线上；通道N11的出口到通道M1的出口的距离、与通道N12的出口到通道M1的出口的距离相同。 In this way, when N is 9 and M is 8, the first layer structure finally forms the (M-1) power of 2, that is, the 7th power of 2, that is, 128 channel outlets, and the second layer structure finally forms the 2 (N-1) power, that is, 2 to the 8th power, that is, 256 channel outlets; that is, it has 128 channels M, and each channel M corresponds to two channels N, with a total of 128 output terminals; for example Channel M1 corresponds to channel N11 and channel N12, and the output directions of channel N11 and channel N12 are opposite and located on the same straight line; the distance from the exit of channel N11 to the exit of channel M1, and the distance from the exit of channel N12 to the exit of channel M1 same. the

在上述实例中，装置的典型特征是集成了128个扩展形喷嘴腔体，例如，如图3所示，第一层结构输入气体，如图2所示，第二层结构输入液体；喷嘴结构如图4所示，每一气体输出通道口对应着两个相对的液体输出通道口；则 气体在喷嘴的最窄点，即喷嘴口，同时也是收集通道的最窄点，具有最大的剪切力，如图5所示，随后在扩展形喷嘴处，产生一个速度递减梯度，利于微泡的聚焦及脱落；因此，利用喷嘴阵列可提高微泡的生产效率。 In the above example, the typical feature of the device is the integration of 128 expanded nozzle cavities, for example, as shown in Figure 3, the first layer of structure input gas, as shown in Figure 2, the second layer of structure input liquid; nozzle structure As shown in Figure 4, each gas output channel opening corresponds to two relative liquid output channel openings; then the gas has the largest shear at the narrowest point of the nozzle, that is, the nozzle opening, which is also the narrowest point of the collecting channel. Force, as shown in Figure 5, then generates a velocity gradient at the expanding nozzle, which is conducive to the focusing and shedding of microbubbles; therefore, the use of nozzle arrays can improve the production efficiency of microbubbles. the

更具体地说，本发明部分例子中，采用了流动聚焦原理(flow-focusing)产生微泡，气体在两侧液体的包被下处于同轴流动中心位置，即中轴线，气体由高速运动的液体聚焦形成稳定的锥体，并且与喷嘴口位置对齐，由于喷嘴口处最窄，具有最大的剪切力，如图5所示，随后的扩展形喷嘴产生一个速度梯度，由锥顶端射出的微射流在喷嘴口处脱落，形成均一尺寸的微泡。 More specifically, in some examples of the present invention, the flow-focusing principle (flow-focusing) is used to generate microbubbles, and the gas is at the center of the coaxial flow under the coating of liquid on both sides, that is, the central axis. The liquid focuses to form a stable cone, which is aligned with the position of the nozzle mouth. Since the nozzle mouth is the narrowest, it has the largest shearing force, as shown in Figure 5, and the subsequent expanding nozzles generate a velocity gradient. The microjets are shed at the nozzle opening, forming microbubbles of uniform size. the

采用上面的各个实施例，就可以将微流控大规模集成芯片用于超声造影剂的制备中，并且，上述各实施例所述集成芯片，还可以应用在制备其他微型的混和物中，例如，微液滴等等。 Using the above embodiments, the microfluidic large-scale integrated chip can be used in the preparation of ultrasound contrast agents, and the integrated chip described in the above embodiments can also be used in the preparation of other miniature mixtures, such as , micro-droplets and so on. the

并且，采用上述任一实施例所述集成芯片，本发明还提供了一种装置。该装置包括第一物质输入单元、第二物质输入单元、贮存单元和至少一如上述任一实施例所述集成芯片，这些集成芯片形成至少一个集成芯片阵列。各集成芯片的收集槽，分别与所述贮存单元相连接，用于收集和导出产物；所述第一物质输入单元分别与各集成芯片的一层结构的各通道1相连接，用于输入第一物质；所述第二物质输入单元分别从各集成芯片的另一层结构的各通道1相连接，用于输入第二物质。该装置可用于制备微米级分散体，包括超声造影剂或其他微型混和物。 Furthermore, the present invention also provides a device using the integrated chip described in any of the above embodiments. The device includes a first substance input unit, a second substance input unit, a storage unit and at least one integrated chip as described in any one of the above embodiments, and these integrated chips form at least one integrated chip array. The collection tanks of each integrated chip are respectively connected with the storage unit for collecting and exporting products; the first substance input unit is respectively connected with eachchannel 1 of the one-layer structure of each integrated chip for inputting the first substance A substance; the second substance input unit is respectively connected to eachchannel 1 of another layer structure of each integrated chip for inputting the second substance. The device can be used to prepare micron-scale dispersions, including ultrasound contrast agents or other micro-mixtures. the

其中，所述第一物质输入单元与所述第二物质输入单元的组合，为气体输入单元与液体输入单元的组合；即所述第一物质输入单元为气体输入单元，所述第二物质输入单元为液体输入单元，或者，所述第一物质输入单元为液体输入单元，所述第二物质输入单元为气体输入单元。或者，所述第一物质输入单元与所述第二物质输入单元均为液体输入单元。 Wherein, the combination of the first substance input unit and the second substance input unit is a combination of a gas input unit and a liquid input unit; that is, the first substance input unit is a gas input unit, and the second substance input unit The unit is a liquid input unit, or, the first substance input unit is a liquid input unit, and the second substance input unit is a gas input unit. Alternatively, both the first substance input unit and the second substance input unit are liquid input units. the

例如，该装置包括气体输入单元、液体输入单元、贮存单元和至少一如上述任一实施例所述集成芯片，这些集成芯片形成至少一个集成芯片阵列。或者说，该装置包括：用于控制气体、液体的流动参数的仪表，以及用于气体及液体通过微喷嘴阵列的芯片，所述微喷嘴具有基本上一致的直径。 For example, the device includes a gas input unit, a liquid input unit, a storage unit and at least one integrated chip as described in any one of the above embodiments, and these integrated chips form at least one integrated chip array. Alternatively, the device includes: instrumentation for controlling flow parameters of gas and liquid, and a chip for passing gas and liquid through an array of micronozzles having substantially uniform diameters. the

例如，整个超声造影剂制备装置的框图如图1所示，气体源输出气体，经流量计或压力计控制流量，输出到大规模微流控芯片中；液体源输出液体，经 数控注射泵控制流量，输出到大规模微流控芯片中；反应生成产品，进行产品收集，例如通过产品收集设备进行收集。例如，通过各集成芯片的收集槽，收集和导出产物，这些收集槽分别与所述贮存单元相连接，将最终的产物导入到所述贮存单元中；这样可以有效地收集最终的产物。 For example, the block diagram of the entire ultrasonic contrast agent preparation device is shown in Figure 1. The gas source outputs gas, which is controlled by a flow meter or pressure gauge, and then output to a large-scale microfluidic chip; the liquid source outputs liquid, which is controlled by a numerically controlled syringe pump. The flow is output to a large-scale microfluidic chip; the reaction generates a product, and the product is collected, for example, collected by a product collection device. For example, the products are collected and exported through the collection tanks of each integrated chip, and these collection tanks are respectively connected with the storage units, and the final products are introduced into the storage units; thus, the final products can be collected effectively. the

一个例子是，所述气体输入单元分别与各集成芯片的一层结构的各通道1相连接，用于输入气体；所述液体输入单元分别从各集成芯片的另一层结构的各通道1相连接，用于输入液体。 An example is that the gas input unit is respectively connected to eachchannel 1 of a layer structure of each integrated chip for inputting gas; the liquid input unit is connected to eachchannel 1 of another layer structure of each integrated chip respectively. Connection for liquid input. the

例如，所述气体输入单元分别与各集成芯片的第一层结构的各通道1相连接，所述液体输入单元分别从各集成芯片的第二层结构的各通道1相连接；或者，所述气体输入单元分别与各集成芯片的第二层结构的各通道1相连接，所述液体输入单元分别从各集成芯片的第一层结构的各通道1相连接。 For example, the gas input unit is respectively connected to eachchannel 1 of the first layer structure of each integrated chip, and the liquid input unit is respectively connected to eachchannel 1 of the second layer structure of each integrated chip; or, the The gas input units are respectively connected to thechannels 1 of the second layer structure of the integrated chips, and the liquid input units are respectively connected to thechannels 1 of the first layer structure of the integrated chips. the

一个例子是，所述气体输入单元包括顺序连接的压力储气罐、减压阀、第一传输管、微流量计、调压阀和第二传输管，所述第二传输管分别与各集成芯片的一层结构的各通道1相连接。 An example is that the gas input unit includes a pressure gas storage tank, a pressure reducing valve, a first delivery pipe, a micro flow meter, a pressure regulating valve and a second delivery pipe connected in sequence, and the second delivery pipe is respectively integrated with each Theindividual channels 1 of the one-layer structure of the chip are connected. the

又一个例子是，所述液体输入单元包括顺序连接的储液器、第三传输管、数字控制式注射泵和第四传输管，所述第四传输管分别与各集成芯片的另一层结构的各通道1相连接。 Yet another example is that the liquid input unit includes a liquid reservoir, a third delivery tube, a digitally controlled syringe pump and a fourth delivery tube connected in sequence, and the fourth delivery tube is connected with another layer of each integrated chip respectively. Eachchannel 1 is connected. the

一个优选的例子是，所述装置的各集成芯片中，各通道截面为矩形，其高度为20μm至30μm；通道M横截面的宽度为30μm至50μm、通道N横截面的宽度为50μm至100μm，或者，通道M横截面的宽度为50μm至100μm、通道N横截面的宽度为30μm至50μm；其中，所述收集通道的结构至少包括以下结构其中之一：外向放大的锥体、球体、半球体或锥台体，其输入端的管径为7μm至25μm，高度为20μm至30μm；或者，圆柱体或长方体，其沿对应的特定通道M输出方向的截面积略小于该特定通道M沿该方向的截面积，高度为20μm至30μm。所述集成芯片的基质为硅、玻璃、聚二甲基硅氧烷、聚甲基丙烯酸甲酯或聚碳酯。第一层结构与第二层结构的组合，为气体通道结构与液体通道结构的组合。H1为1，H2至HN均为2；N为8，M为7，第一层结构和第二层结构均在收集通道两侧对称设有两个初始通道，即2个气体入口和2个液体入口，则第一层结构最终形成2个2的(M-1)次方，即128个通道出口，第二层结构最终形成2个2的(N-1)次方，即256个通道出口；该实例中，装置的典型 特征是集成了128个扩展形喷嘴腔体，气体在喷嘴的最窄点，即收集通道的最窄点，具有最大的剪切力，随后的扩展形喷嘴产生一个速度递减梯度，利于微泡的聚焦及脱落；喷嘴阵列可提高微泡的生产效率。 A preferred example is that, in each integrated chip of the device, each channel has a rectangular section with a height of 20 μm to 30 μm; the width of the channel M cross section is 30 μm to 50 μm, and the width of the channel N cross section is 50 μm to 100 μm, Alternatively, the width of the channel M cross-section is 50 μm to 100 μm, and the width of the channel N cross-section is 30 μm to 50 μm; wherein, the structure of the collection channel includes at least one of the following structures: outwardly enlarged cones, spheres, and hemispheres Or a frustum, the diameter of the input end is 7 μm to 25 μm, and the height is 20 μm to 30 μm; or, a cylinder or a cuboid, the cross-sectional area along the output direction of the corresponding specific channel M is slightly smaller than that of the specific channel M along this direction The cross-sectional area and the height are 20 μm to 30 μm. The substrate of the integrated chip is silicon, glass, polydimethylsiloxane, polymethyl methacrylate or polycarbonate. The combination of the first layer structure and the second layer structure is the combination of the gas channel structure and the liquid channel structure. H1 is 1, H2 to HN are 2; N is 8, M is 7, the first layer structure and the second layer structure are symmetrically provided with two initial channels on both sides of the collection channel, that is, 2 gas inlets and 2 The liquid inlet, the first layer structure finally forms two 2 (M-1) powers, that is, 128 channel outlets, and the second layer structure finally forms two 2 (N-1) powers, that is, 256 channels Outlet; in this example, the typical feature of the device is the integration of 128 expanded nozzle cavities, the gas has the largest shear force at the narrowest point of the nozzle, that is, the narrowest point of the collection channel, and the subsequent expanded nozzle produces A gradient of decreasing speed facilitates the focusing and shedding of microbubbles; the nozzle array can improve the production efficiency of microbubbles. the

更具体地说，如图1所示，气体由压力罐经减压阀输送到芯片的气体入口，由微流量计、调压阀监控气体的流动，连续性的液体由数字控制式注射泵以恒定的流速泵入液体入口，经过微流控大规模集成芯片产生的微泡由特殊的贮存瓶存储，该贮存瓶为现有技术。 More specifically, as shown in Figure 1, the gas is delivered from the pressure tank to the gas inlet of the chip through the pressure reducing valve. A constant flow rate is pumped into the liquid inlet, and the microbubbles generated by the microfluidic LSI chip are stored in a special storage bottle, which is the prior art. the

其中，微流控大规模集成芯片是利用微纳米加工技术制作而成，例如，该芯片具有上下两层结构。 Among them, the microfluidic large-scale integrated chip is manufactured by using micro-nano processing technology, for example, the chip has a structure of upper and lower layers. the

更具体的一个例子是，该芯片底层结构如图2所示。通道由两个对称设置液体入口开始逐级扩展，最终经过8级梯度，共7级扩展形成256个分支，即N为8。每两个属于一个初始入口的分支作为微喷嘴的液体入口，这样就形成128个微喷嘴。图2中点划线指示的位置a′是一个微喷嘴的气体入口，共128个气体入口，产生的微泡经收集槽收集存储。 A more specific example is that the underlying structure of the chip is shown in FIG. 2 . The channel expands step by step from two symmetrically arranged liquid inlets, and finally passes through 8-level gradients, and a total of 7-level expansion forms 256 branches, that is, N is 8. Every two branches belonging to an initial inlet serve as the liquid inlets of the micro-nozzles, thus forming 128 micro-nozzles. The position a' indicated by the dotted line in Fig. 2 is the gas inlet of a micronozzle, a total of 128 gas inlets, and the generated microbubbles are collected and stored in the collection tank. the

该芯片顶层通道结构如图3所示，与底层结构类似，通道由两个对称设置气体入口开始逐级扩展，有7个梯度，经6级扩展，即M为7，单个初始气体入口最终形成64个分支。因此，经过扩展总共形成了128个分支，与底层相对应的气128个气体入口相通，如a与a′相通。 The channel structure on the top layer of the chip is shown in Figure 3. Similar to the bottom layer structure, the channel starts to expand step by step from two symmetrically arranged gas inlets. There are 7 gradients, and after 6 stages of expansion, that is, M is 7, and a single initial gas inlet is finally formed. 64 branches. Therefore, a total of 128 branches have been formed after expansion, communicating with 128 gas inlets corresponding to the bottom layer, such as a communicating with a'. the

其中，可以将底层与顶层的结构互换，如图6所示，这是本发明中微流控大规模集成芯片的替换形式，顶层为液体通道结构，底层主要为气体通道，底层的两个初始入口经6级扩展成128个分支，作为微喷嘴的气体入口。图7为图6中虚线框所示部分一个微喷嘴的放大图，与图4的主要区别是两个液体入口与顶层液体通道相通。 Among them, the structure of the bottom layer and the top layer can be interchanged, as shown in Figure 6, this is the replacement form of the microfluidic large-scale integrated chip in the present invention, the top layer is a liquid channel structure, the bottom layer is mainly a gas channel, and the two bottom layers The initial inlet is expanded into 128 branches through 6 stages, which are used as the gas inlet of the micro-nozzle. Fig. 7 is an enlarged view of a micro-nozzle shown in the dotted line box in Fig. 6, and the main difference from Fig. 4 is that the two liquid inlets communicate with the liquid channel on the top layer. the

如图4所示，是图2虚线所示部分的放大图，为该集成芯片阵列中一个微喷嘴的示意图，微喷嘴是微泡形成的基本单元，其中，箭头代表气体、液体的流动方向，一个气体入口与顶层气体通道相通，在大箭头形状的收集通道的最窄处剪切力最大。收集通道入口的口径在7μm-25μm范围内，通道高度为25μm，优选的是与同一层结构内其他通道的高度一致，气体入口宽度为30μm-50μm，液体入口宽度为50μm-100μm，气体在两侧高速流动的液体聚焦作用下形成位于中轴线位置的稳定锥形，如图5所示，并且与喷嘴口位置对齐，由于喷嘴口处 最窄，具有最大的剪切力，随后的扩展形喷嘴产生一个速度梯度，由锥顶端射出的微射流在喷嘴口处脱落，形成均一尺寸的微泡，微泡形成示意图如图5所示，可以形象地看到流动聚焦形成的锥形及微泡。 As shown in Figure 4, it is an enlarged view of the part shown in dotted line in Figure 2, which is a schematic diagram of a micronozzle in the integrated chip array, the micronozzle is the basic unit of microbubble formation, wherein the arrow represents the flow direction of gas and liquid, A gas inlet communicates with the top gas channel, where the shear force is greatest at the narrowest point of the large arrow-shaped collection channel. The diameter of the inlet of the collection channel is in the range of 7 μm-25 μm, the height of the channel is 25 μm, preferably consistent with the height of other channels in the same layer structure, the width of the gas inlet is 30 μm-50 μm, and the width of the liquid inlet is 50 μm-100 μm. Under the action of the high-speed flowing liquid on the side, a stable cone is formed on the central axis, as shown in Figure 5, and is aligned with the position of the nozzle opening. Since the nozzle opening is the narrowest and has the largest shear force, the subsequent expanding nozzle A velocity gradient is generated, and the microjet ejected from the tip of the cone falls off at the nozzle mouth to form microbubbles of uniform size. the

其中，如图9所示，本装置的微喷嘴形状可以由形成速度梯度的其它几何形状代替，如矩形结构，只需能够产生一个速度梯度，让锥顶端射出的微射流在喷嘴口处脱落即可。 Wherein, as shown in Figure 9, the shape of the micro-nozzle of the device can be replaced by other geometric shapes that form a velocity gradient, such as a rectangular structure, as long as a velocity gradient can be produced, the micro-jet ejected from the cone tip will fall off at the nozzle opening. Can. the

需要说明的是，各图中对于部分尺寸进行放大，这是出于图解说明的目的，而没有按照严格的比例进行绘制。这些图示的不严格绘制，对本发明各技术方案和各实施例均不构成任何额外限制。 It should be noted that, in each figure, some dimensions are enlarged for the purpose of illustration, and are not drawn in strict proportion. These illustrations are not strictly drawn, and do not constitute any additional restrictions on the technical solutions and embodiments of the present invention. the

采用上面的各个实施例，就可以大规模集成扩展形微喷嘴阵列，通过在不同的通道入口输入气体和液体，可以实现并且有利于微泡的大规模生产。采用各种不同的气体和液体，从集成芯片的各组结构的第一层结构各通道1中输入气体，本发明的装置可用于制备多种类型的超声造影剂微泡。 Using the above embodiments, the extended micro-nozzle array can be integrated on a large scale, and the large-scale production of micro-bubbles can be realized and facilitated by inputting gas and liquid at different channel inlets. Using various gases and liquids, the gas is input from eachchannel 1 of the first layer structure of each group structure of the integrated chip, and the device of the present invention can be used to prepare various types of ultrasonic contrast agent microbubbles. the

交换气体、液体入口，即从集成芯片的各组结构的第一层结构各通道1中输入液体，可用于生成大量微液滴；或者，分别通入两种不相容的液体，会生成底层液体包裹上层液体的微液滴，非常适用于食品行业和化妆品行业。 Exchange gas and liquid inlet, that is, input liquid from eachchannel 1 of the first layer structure of each group structure of the integrated chip, which can be used to generate a large number of micro-droplets; or, two incompatible liquids are respectively introduced to generate the bottom layer Micro-droplets of liquid enveloping the upper liquid, very suitable for the food industry and cosmetic industry. the

其中，采用多个收集通道所形成的微喷嘴阵列，微喷嘴的数量可以增减，对于一组结构，最少有一个喷嘴，第一层结构设置N级梯度时，通道的扩展级数为N-1，即比梯度数N少1。在只有一个初始入口的情况下，最后分支是2N-1个，喷嘴的个数一般是2N-2个。在有两个对称设置的初始入口的情况下，喷嘴的个数一般为2N-1个。例如，N为8时，有8级梯度，扩展了7级，每级扩展一倍，第一级梯度设置1个通道1，第八级梯度设置128个通道8，在只有一个初始入口的情况下，对应有64个喷嘴；在有两个对称设置的初始入口的情况下，对应有128个喷嘴。在易于控制流体流速、气体压力的前提下，喷嘴个数越多越好。 Among them, the number of micro-nozzles can be increased or decreased by using a micro-nozzle array formed by multiple collection channels. For a group of structures, there is at least one nozzle. 1, which is 1 less than the gradient number N. In the case of only one initial inlet, the number of final branches is 2N-1, and the number of nozzles is generally 2N-2. In the case of two symmetrically arranged initial inlets, the number of nozzles is generally 2N-1. For example, when N is 8, there are 8 levels of gradients, 7 levels are expanded, and each level is doubled. The first level of gradients is set to 1channel 1, and the eighth level of gradients is set to 128channels 8. In the case of only one initial entry Below, there are 64 nozzles; in the case of two symmetrically arranged initial inlets, there are 128 nozzles. On the premise that it is easy to control the fluid flow rate and gas pressure, the more nozzles the better. the

并且，还提供了一种制备微米级分散体的方法，这些微米级分散体之间不相溶；该方法应用于一包括集成芯片阵列的装置中，所述集成芯片阵列由至少一如上述任一实施例所述集成芯片组成；所述装置为上述任一实施例所述装置。所述方法包括以下步骤。 And, also provide a kind of method for preparing micron-scale dispersion, these micron-scale dispersions are immiscible; This method is applied in a device that comprises integrated chip array, and described integrated chip array is made of at least one as above-mentioned any The integrated chip described in one embodiment is composed; the device is the device described in any one of the above embodiments. The method includes the following steps. the

A1、按第一预设置条件，向各集成芯片的一层结构的各通道1输入第一物 质；按第二预设置条件，向各集成芯片的另一层结构的各通道1输入第二物质。其中，第一物质和第二物质可以为不同的液体；或者，第一物质可以为液体，第二物质可以为气体；或者，第一物质可以为气体，第二物质可以为液体。 A1. According to the first preset condition, input the first substance to eachchannel 1 of the one-layer structure of each integrated chip; according to the second preset condition, input the second substance to eachchannel 1 of another layer structure of each integrated chip substance. Wherein, the first substance and the second substance may be different liquids; or, the first substance may be liquid, and the second substance may be gas; or, the first substance may be gas, and the second substance may be liquid. the

一个例子是，步骤A1中，向各集成芯片的一层结构的各通道1输入气体，压力低于10psi，压力大于0即可，向各集成芯片的另一层结构的各通道1输入液体，流速低于3μL/s，流速大于0即可；例如，输入气体的压力低于5psi，输入液体的流速低于1.5μL/s；又如，输入气体的压力低于8psi，输入液体的流速低于2.5μL/s；又如，输入气体的压力低于7.5psi，输入液体的流速低于2.0μL/s；又如，输入气体的压力低于4psi，输入液体的流速低于2.8μL/s。这样，就可以制备均一的微泡，这些微泡可应用于超声造影剂中。 An example is that in step A1, gas is input to eachchannel 1 of the one-layer structure of each integrated chip, the pressure is lower than 10 psi, and the pressure is greater than 0, and liquid is input to eachchannel 1 of another layer structure of each integrated chip, The flow rate is lower than 3μL/s, and the flow rate is greater than 0; for example, the pressure of the input gas is lower than 5psi, and the flow rate of the input liquid is lower than 1.5μL/s; another example, the pressure of the input gas is lower than 8psi, and the flow rate of the input liquid is low less than 2.5μL/s; as another example, the pressure of the input gas is lower than 7.5psi, and the flow rate of the input liquid is lower than 2.0μL/s; another example, the pressure of the input gas is lower than 4psi, and the flow rate of the input liquid is lower than 2.8μL/s . In this way, uniform microbubbles can be prepared, which can be used in ultrasound contrast agents. the

具体应用中，所述气体至少包括氮气、氟碳类气体、氟硫类气体其中之一；所述液体至少包括磷脂类液体、表面活性剂类液体、上述液体的改性液其中之一。 In a specific application, the gas includes at least one of nitrogen, fluorocarbon gas, and fluorine-sulfur gas; the liquid includes at least one of a phospholipid liquid, a surfactant liquid, and a modified liquid of the above liquids. the

为了保证微泡在体内具有较长的存活时间，该装置可使用的气体材料主要是在血浆中有低溶解度的氟碳、氟硫类气体，如C3F8、C4F10、SF6等，也可以是这些气体成分之间的组合或者它们与氮气的组合如氮气和C4F10、氮气和C3F8的组合。 In order to ensure that the microbubbles have a longer survival time in the body, the gas materials that can be used in this device are mainly fluorocarbons and fluorine-sulfur gases with low solubility in plasma, such as C3F8, C4F10, SF6, etc. These gases can also be used Combinations between components or their combinations with nitrogen such as nitrogen and C4F10, nitrogen and C3F8. the

而制备液泡，即微泡，其壳材来自液体，这些液体主要包括磷脂类，如DPPC、DPPA、DPPE、DSPC、DSPE、DSPA等；表面活性剂类，如Span20、Span60、Span 80等，Tween20、Tween 60、Tween 80等，以及磷脂类的组合、表面活性剂类的组合、或者磷脂类与表面活性剂类的组合；这些材料既能显著降低气液界面的表面张力，又具有良好的生物相容性和血液相容性。 To prepare vacuoles, i.e. microbubbles, the shell material comes from liquids, these liquids mainly include phospholipids, such as DPPC, DPPA, DPPE, DSPC, DSPE, DSPA, etc.; surfactants, such as Span20, Span60, Span 80, etc., Tween20 , Tween 60, Tween 80, etc., and the combination of phospholipids, surfactants, or phospholipids and surfactants; these materials can not only significantly reduce the surface tension of the gas-liquid interface, but also have good biological properties compatibility and blood compatibility. the

或者，制备液泡的壳材是在上述壳材的基础上进行改性得到的具有特殊功能的材料，包括DSPE-PEG2000、DSPE-PEG5000、DSPE-PEG(2000)Biotin、DSPE-PEG(2000)Carboxylic Acid等等。磷脂的疏水性长链和亲水性极性头的分子结构使其在水性环境中能够自行调整其分子结构，疏水性链紧密排列在一起，而亲水性头暴露于水相，最终形成囊泡状结构，这种结构能够阻碍气体扩散出形成的微泡，亲水性的外壳使其具有优良的生物相容性。同时，不同的极性头可以使磷脂具有不同的荷电性，通过调整这些荷电磷脂的成分和比例，可以调整磷脂微泡的稳定性。含有PEG长链的磷脂，如DSPE-PEG2000，具有抵抗其 在体内循环时被网状内皮系统捕获的能力，延长在体内的循环的时间。含有Biotin配体的磷脂，如DSPE-PEG(2000)Biotin，具有识别抗体avidin能力，因此具有靶向功能。这样，通过不同的磷脂组分之间进行组合，并调整各组分的比例，就可以得到不同功能和作用的微气泡。 Alternatively, the shell material for preparing the vacuole is a material with special functions obtained by modifying the above shell material, including DSPE-PEG2000, DSPE-PEG5000, DSPE-PEG (2000) Biotin, DSPE-PEG (2000) Carboxylic Acid and so on. The molecular structure of phospholipids' long hydrophobic chains and hydrophilic polar heads enables them to self-adjust their molecular structure in an aqueous environment. The hydrophobic chains are closely packed together, while the hydrophilic heads are exposed to the aqueous phase, eventually forming vesicles. Bubble-like structure, which can hinder the diffusion of gas to form microbubbles, and the hydrophilic shell makes it have excellent biocompatibility. At the same time, different polar heads can make phospholipids have different charges, and by adjusting the composition and ratio of these charged phospholipids, the stability of phospholipid microbubbles can be adjusted. Phospholipids containing PEG long chains, such as DSPE-PEG2000, have the ability to resist being captured by the reticuloendothelial system during circulation in the body, prolonging the time of circulation in the body. Phospholipids containing Biotin ligands, such as DSPE-PEG (2000) Biotin, have the ability to recognize antibody avidin, so they have targeting functions. In this way, by combining different phospholipid components and adjusting the ratio of each component, microbubbles with different functions and effects can be obtained. the

A2、调节第一物质和/或第二物质的输入条件，使各个第一层结构的各通道M输出的物质以同轴流动形式，由对应的各特定通道N的物质按轴向包裹通道M输出的物质，分别形成待收集产物流，其轴向与所述垂线重合。 A2. Adjust the input conditions of the first substance and/or the second substance, so that the substances output by each channel M of each first layer structure flow coaxially, and the corresponding specific channel N wraps the channel M axially The output materials respectively form the product stream to be collected, the axis of which coincides with the vertical line. the

例如，步骤A2中，单独调节不同气体或不同液体的输入条件，或者单独调节气体或液体的输入条件，或者同时调节气体和液体的输入条件，使各个第一层结构的各通道M输出的物质以同轴流动形式，由对应的各特定通道N的物质按轴向进行包裹对应通道M输出的物质，分别形成待收集产物流，其轴向与所述垂线重合；例如，分别形成一倒锥体的待收集产物流，其轴向与所述垂线重合。 For example, in step A2, the input conditions of different gases or different liquids are adjusted separately, or the input conditions of gases or liquids are adjusted separately, or the input conditions of gases and liquids are adjusted simultaneously, so that the substances output by each channel M of each first layer structure In the form of coaxial flow, the material output by the corresponding channel M is wrapped axially by the material corresponding to each specific channel N, respectively forming a product flow to be collected, and its axial direction coincides with the vertical line; for example, forming an inverted The product stream to be collected of the cone has its axis coincident with said vertical line. the

A3、从各收集通道的输出端收集和导出产物。 A3. Collect and export products from the output ends of each collection channel. the

例如，向各集成芯片的第一层结构的各通道1输入气体，向各集成芯片的第二层结构的各通道1输入液体；这样，可以得到一种通过气体由两侧液体流动聚焦在喷嘴处形成微泡的方法，形成高度单分散的微泡，适合于超声造影成像技术。或者，向各集成芯片的第一层结构的各通道1输入液体，向各集成芯片的第二层结构的各通道1输入气体；这样，可以得到一种通过液体由两侧气体流动聚焦在喷嘴处形成微液滴的方法。 For example, the gas is input to eachchannel 1 of the first layer structure of each integrated chip, and the liquid is input to eachchannel 1 of the second layer structure of each integrated chip; like this, it is possible to obtain a gas flow focused on the nozzle by the liquid flow on both sides. The method of forming microbubbles at the same place forms highly monodisperse microbubbles, which is suitable for ultrasound contrast imaging technology. Or, input liquid to eachchannel 1 of the first layer structure of each integrated chip, and input gas to eachchannel 1 of the second layer structure of each integrated chip; like this, can obtain a kind of through liquid and focus on the nozzle by the gas flow of both sides methods for the formation of microdroplets. the

同上所述，一个优选的例子是，所述集成芯片，H1为1，H2至HN均为2，N为9，M为8。其具有128个通道M，每个通道M对应着两个通道N，共有128个输出端；例如通道M1对应着通道N11和通道N12，通道N11和通道N12的输出方向相对，并位于同一直线上；通道N11的出口到通道M1的出口的距离、与通道N12的出口到通道M1的出口的距离相同。 As mentioned above, a preferred example is that, in the integrated chip, H1 is 1, H2 to HN are all 2, N is 9, and M is 8. It has 128 channels M, each channel M corresponds to two channels N, and has a total of 128 output terminals; for example, channel M1 corresponds to channel N11 and channel N12, and the output directions of channel N11 and channel N12 are opposite and located on the same straight line ; The distance from the exit of the channel N11 to the exit of the channel M1 is the same as the distance from the exit of the channel N12 to the exit of the channel M1. the

另一个例子是，步骤A1中，向各集成芯片的第一层结构的各通道1输入气体，向各集成芯片的第二层结构的各通道1输入液体；步骤A2中，使各个第一层结构的各通道M输出的气体以同轴流动形式，由各个第二层结构的对应某一通道M的各通道N，所流出的液体按轴向包裹对应该通道M的气体，分别形成待收集微球流。其中，同轴流动形式已如前述。 Another example is that in step A1, gas is input to eachchannel 1 of the first layer structure of each integrated chip, and liquid is input to eachchannel 1 of the second layer structure of each integrated chip; in step A2, each first layer The gas output by each channel M of the structure is in the form of coaxial flow, and the liquid flowing out from each channel N corresponding to a certain channel M of each second layer structure wraps the gas corresponding to the channel M in the axial direction, respectively forming microsphere flow. Wherein, the coaxial flow form is as mentioned above. the

在上例的基础之上，步骤A2中，各个第一层结构的各通道M输出的气体，分别在两侧高速流动的液体聚焦作用下形成中轴线位于所述垂线位置的稳定倒锥体，各倒锥体的尖端分别与各收集通道位置相对。 On the basis of the above example, in step A2, the gas output from each channel M of each first layer structure forms a stable inverted cone with the central axis at the position of the vertical line under the focusing action of the high-speed flowing liquid on both sides. , the tip of each inverted cone is opposite to the position of each collection channel. the

又一个例子是，步骤A1中，向各集成芯片的第一层结构的各通道1输入第一液体，向各集成芯片的第二层结构的各通道1输入气体或第二液体；步骤A2中，使各个第一层结构的各通道M输出的第一液体以同轴流动形式，被气体或第二液体按轴向包裹，分别形成待收集微液滴流；例如分别形成一倒锥体的待收集微液滴流。 Another example is that in step A1, the first liquid is input to eachchannel 1 of the first layer structure of each integrated chip, and the gas or the second liquid is input to eachchannel 1 of the second layer structure of each integrated chip; in step A2 , so that the first liquid output by each channel M of each first layer structure is in the form of coaxial flow, wrapped by gas or second liquid in the axial direction, respectively forming micro-liquid droplet flow to be collected; for example, forming an inverted cone respectively Microfluidic droplet stream to be collected. the

在上述各例中，步骤A1中，所述液体可以预先加入特异性配体；或者，步骤A3之后，向所述产物加入特异性配体。这样，可以根据需要，加入不同的特异性配体，从而制备出靶向超声造影剂。 In each of the above examples, in step A1, specific ligands may be added to the liquid in advance; or, after step A3, specific ligands may be added to the product. In this way, different specific ligands can be added according to needs, so as to prepare targeted ultrasound contrast agents. the

下面继续对集成芯片、装置和方法，尤其是制备超声造影剂的方法，做出详细说明。 The integrated chip, device and method, especially the method for preparing ultrasound contrast agent will be described in detail below. the

在超声造影成像技术中，微泡造影剂的尺寸影响它通过肺部微循环的能力及对超声的反射率，直径必须小于7μm才可以安全通过肺部微动脉而不引起堵塞，其中，超声的散射强度、入射强度具有以下关系公式(1)： In contrast-enhanced ultrasound imaging technology, the size of the microbubble contrast agent affects its ability to pass through the pulmonary microcirculation and its reflectivity to ultrasound. The diameter must be less than 7 μm to safely pass through the pulmonary arterioles without causing blockage. Among them, the ultrasound Scattering intensity and incident intensity have the following relationship formula (1):

$\mathrm{<span><span>I</span>I</span>}<span><span>/</span>/</span>{\mathrm{<span><span>I</span>I</span>}}_{\mathrm{<span><span>0</span>0</span>}}<span><span>~</span>~</span>\frac{\mathrm{<span><span>1</span>1</span>}}{\mathrm{<span><span>9</span>9</span>}}\mathrm{<span><span>nV</span>nV</span>}<span><span>[</span>[</span>{\mathrm{<span><span>k</span>k</span>}}^{\mathrm{<span><span>4</span>4</span>}}{\mathrm{<span><span>r</span>r</span>}}^{\mathrm{<span><span>6</span>6</span>}}{<span><span>(</span>(</span>{\mathrm{<span><span>\&gamma;</span>\&gamma;</span>}}_{\mathrm{<span><span>c</span>c</span>}}<span><span>+</span>+</span>{\mathrm{<span><span>\&gamma;</span>\&gamma;</span>}}_{\mathrm{<span><span>d</span>d</span>}}\mathrm{<span><span>cos</span>cos</span>}\mathrm{<span><span>\&theta;</span>\&theta;</span>}<span><span>)</span>)</span>}^{\mathrm{<span><span>2</span>2</span>}}<span><span>/</span>/</span>{\mathrm{<span><span>d</span>d</span>}}^{\mathrm{<span><span>2</span>2</span>}}<span><span>]</span>]</span><span><span>-</span>-</span><span><span>-</span>-</span><span><span>-</span>-</span><span><span>(</span>(</span>\mathrm{<span><span>1</span>1</span>}<span><span>)</span>)</span>$

其中，I、I₀分别是超声的散射强度、入射强度；n是散射粒子数；V是散射体积；k是波数；r是粒子半径，即微泡半径；γ_c是压缩项(γ_c＝(k_s-k_m)/k_m，k_s、k_m分别是散射粒子的压缩率和介质的压缩率，公知技术)；γ_d是密度项(γ_d＝(3ρ_s-3ρ_m)/(2ρ_s+ρ_m)，ρ_s、ρ_m分别是散射粒子和介质的密度，公知技术)；θ是散射角；d是到散射粒子的距离。由公式(1)可以看出，微泡的散射率是微泡半径六次方的函数，即，超声造影剂微泡的散射强度与气泡半径六次方成正比，说明微泡越小，反射率越差，所以微泡的最佳直径尺寸不能太小。因此，临床应用要求微泡的最佳直径尺寸在2μm至5μm之间。 Wherein, I, I₀ are the scattering intensity and incident intensity of ultrasound respectively; n is the number of scattering particles; V is the scattering volume; k is the wave number;_r is the particle radius, i.e. the microbubble radius_; (k_s -k_m )/_km , k_s and_km are the compressibility of the scattering particles and the medium respectively, known technology); γ_d is the density term (γ_d =(3ρ_s -3ρ_m )/ (2ρ_s +ρ_m ), ρ_s and ρ_m are the densities of the scattering particles and the medium respectively, known technology); θ is the scattering angle; d is the distance to the scattering particles. It can be seen from the formula (1) that the scattering rate of the microbubble is a function of the sixth power of the microbubble radius, that is, the scattering intensity of the microbubble of the ultrasound contrast agent is proportional to the sixth power of the bubble radius, indicating that the smaller the microbubble, the more reflective The worse the rate is, so the optimal diameter size of the microbubbles cannot be too small. Therefore, clinical applications require the optimal diameter of microbubbles to be between 2 μm and 5 μm.

由微流控聚焦系统产生的微泡直径主要与气体、液体的流动速率有关，表面张力的影响较小，可以忽略，如以下公式(2)所示： The diameter of microbubbles produced by the microfluidic focusing system is mainly related to the flow rate of gas and liquid, and the influence of surface tension is small and can be ignored, as shown in the following formula (2):

d_b/D∝(Q_g/Q_l)^0.4                            (2) d_b /D∝(Q_g /Q_l )^0.4 (2)

其中，Q_g、Q_l分别是气体、液体的流动速率；D是喷嘴口直径；Q_g/Q_l＜1。 Wherein, Q_g and Q_l are the flow rates of gas and liquid respectively; D is the diameter of the nozzle opening; Q_g /Q_l <1.

利用调压阀调整气体压力，设置注射泵流速参数控制液体的流动速率，一般情况下气体压力P＜5psi(Pounds per square inch，1psi＝6.895kPa)，液体流速Q＜1.5μL/s，可以得到直径d_b＜5μm的微泡，微泡粒径的多分散性指数＜2％。 Use the pressure regulating valve to adjust the gas pressure, and set the flow rate parameters of the syringe pump to control the flow rate of the liquid. Generally, the gas pressure P<5psi (Pounds per square inch, 1psi=6.895kPa), and the liquid flow rate Q<1.5μL/s, can be obtained For microbubbles with a diameter d_b < 5 μm, the polydispersity index of the microbubble particle size is < 2%.

为了证实本发明装置制备的微泡的粒径及生产效率，采用上述一集成芯片进行了以下实验，其在每一层结构为对称两个初始入口，H1为1，H2至HN均为2，N为8，M为7，第一层结构有128个M通道，第二层结构有256个N通道；其集成了128个扩展形喷嘴腔体，第一层结构通入气体，在喷嘴的最窄点输出，如图4、图5所示。具体说明如下。 In order to confirm the particle size and production efficiency of the microbubbles prepared by the device of the present invention, the above-mentioned integrated chip was used to carry out the following experiments. It has two symmetrical initial entrances in each layer structure, H1 is 1, H2 to HN are 2, N is 8, M is 7, the first layer structure has 128 M channels, and the second layer structure has 256 N channels; it integrates 128 expanded nozzle cavities, the first layer structure passes through the gas, and the nozzle The narrowest point output, as shown in Figure 4 and Figure 5. The details are as follows. the

实验一： experiment one:

气体采用氮气，液体采用如下配制：PH＝7.4的磷酸盐缓冲溶液(PBS)8ml，吐温80(Tween80)1ml配制成均匀的混合物，调节气体压力为1.8psi，液体流动速率2.0μLs^-1，可获得微泡直径约为3.6μm，每分钟大约可以产生7×109个微泡。如图8所示，是在奥林巴斯倒置显微镜下拍到的产生的微泡的图片，可以看出微泡粒径分布具有高度单分散性，粒径分布比较均匀，外壳厚度员比较均匀。 The gas is nitrogen, and the liquid is prepared as follows: Phosphate buffer solution (PBS) 8ml of pH = 7.4, Tween 80 (Tween80) 1ml to prepare a uniform mixture, the gas pressure is adjusted to 1.8psi, the liquid flow rate is 2.0μLs^-1 , The obtained microbubble diameter is about 3.6 μm, and about 7×109 microbubbles can be produced per minute. As shown in Figure 8, it is a picture of the generated microbubbles taken under an Olympus inverted microscope. It can be seen that the particle size distribution of the microbubbles is highly monodisperse, the particle size distribution is relatively uniform, and the thickness of the shell is relatively uniform. .

实验二：在实验一的基础上，液体流动速率不变，增加气体压力至4.5psi，获得微泡直径约为6μm，从显微镜得到的结果来看，粒径分布同样比较均匀，比较实验一与实验二，可以得到：改变气体压力可灵活控制微泡粒径的大小；同理，当然也可改变液体流速，可达到相同的效果。 Experiment 2: On the basis ofExperiment 1, the liquid flow rate remains unchanged, and the gas pressure is increased to 4.5psi to obtain a microbubble diameter of about 6μm. From the results obtained by the microscope, the particle size distribution is also relatively uniform. CompareExperiment 1 with In the second experiment, it can be obtained that changing the gas pressure can flexibly control the size of the microbubble particle size; in the same way, of course, the liquid flow rate can also be changed to achieve the same effect. the

实验三：在实验一的基础上，气体压力不变，增加液体流速至2.7μLs^-1，获得微泡直径约为3μm，粒径分布同样比较均匀，比较实验二与实验三，可以得到：可改变液体流速可灵活控制微泡粒径的大小。 Experiment 3: On the basis ofExperiment 1, the gas pressure remains unchanged, and the liquid flow rate is increased to 2.7μLs^-1 , the diameter of microbubbles is about 3μm, and the particle size distribution is also relatively uniform. ComparingExperiment 2 and Experiment 3, it can be obtained: Changing the liquid flow rate can flexibly control the size of the microbubble particle size.

实验四：气体采用全氟碳(PFC)，液体采用如下方法配制：脂质二棕榈酸磷脂酰胆碱(DPPC)、二棕榈磷脂酸聚乙二醇(DPPA)、5000化二棕榈酸磷脂酰乙醇胺(DPPE-PEG5000)以81∶8∶10的摩尔比，溶解在氯仿(CHCl3)中，在真空及氮气条件下形成均匀的混合物，加入1mol％的荧光剂(DiI-C18)和4mg/ml的NaCl溶液到含有磷脂混合物的试管中，室温下超声处理20分钟并混入10％浓度的丙三醇和1、2丙二醇的混合物(GPW)，气体压力10psi，液体流动速率1.0μLs^-1，通过上述芯片可获得微泡直径5μm，每分钟大约可以产生8×109个微 泡。 Experiment 4: The gas is made of perfluorocarbon (PFC), and the liquid is prepared by the following method: lipid dipalmitate phosphatidylcholine (DPPC), dipalmitate phosphatidic acid polyethylene glycol (DPPA), 5000 liters of dipalmitate phosphatidylcholine Ethanolamine (DPPE-PEG5000) was dissolved in chloroform (CHCl3) with a molar ratio of 81:8:10, and a homogeneous mixture was formed under vacuum and nitrogen conditions, and 1mol% fluorescent agent (DiI-C18) and 4mg/ml Put the NaCl solution into the test tube containing the phospholipid mixture, sonicate it for 20 minutes at room temperature and mix it with a mixture of 10% glycerol and 1,2 propylene glycol (GPW), the gas pressure is 10psi, the liquid flow rate is 1.0μLs^-1 , through the above The chip can obtain microbubbles with a diameter of 5 μm, and can generate about 8×109 microbubbles per minute.

本发明具有以下优点： The present invention has the following advantages:

最显著的优点是制作的超声造影剂微泡具有高度单分散性和粒径可控性，满足超声造影成像技术的要求。微泡粒径的多分散性指数＜2％，粒径随着气体压力的增加而变大，随着液体流速的增加而减小，控制非常灵活。并且，通过采用微喷嘴阵列，大大提高了微泡的制备效率；同时，该装置具有重复使用性，降低了生产成本。 The most notable advantage is that the produced ultrasound contrast agent microbubbles have high monodispersity and particle size controllability, which meets the requirements of ultrasound contrast imaging technology. The polydispersity index of microbubble particle size is less than 2%. The particle size increases with the increase of gas pressure and decreases with the increase of liquid flow rate, and the control is very flexible. Moreover, by adopting the micro-nozzle array, the preparation efficiency of the micro-bubble is greatly improved; at the same time, the device is reusable and the production cost is reduced. the

本发明的装置可用于制备多种类型的超声造影剂微泡，制备的微泡外壳厚度均匀。 The device of the invention can be used to prepare various types of ultrasonic contrast agent microbubbles, and the prepared microbubble shells have uniform thickness. the

常规的超声造影剂可直接用该装置制备，如脂类、白蛋白类、多聚体类、表面活性剂类等；如上所述，靶向超声造影剂的制备，有两种选择，一种是目前最常用的：在常规超声造影剂制备完成后加入特异性配体，另行制备；另一种是在造影剂制备完成前的制备液中预先加入特异性配体，在形成造影剂微泡的同时特异性配体就会镶嵌到微泡的壳上，有些壳材如蛋白分子在高温和超声条件下失活，所以这种方法是以往声振法所不能的。该发明装置对这两种方法都适用，特别是当采用后一种选择时更具有优势，不仅减少了制备环节，还减少了制备环节微泡的破坏。 Conventional ultrasound contrast agents can be directly prepared with this device, such as lipids, albumins, polymers, surfactants, etc.; as mentioned above, there are two options for the preparation of targeted ultrasound contrast agents, one It is the most commonly used at present: add specific ligands after the preparation of conventional ultrasound contrast agents and prepare them separately; At the same time, specific ligands will be embedded on the shell of microbubbles, and some shell materials, such as protein molecules, are inactivated under high temperature and ultrasonic conditions, so this method cannot be used in the previous acoustic vibration method. The device of the invention is applicable to both methods, especially when the latter option is adopted, it has more advantages, not only reducing the preparation process, but also reducing the destruction of microbubbles in the preparation process. the

并且，本发明装置和方法在造影剂制备过程中产热少，因此尤其适合用于制备兼药物或基因靶向载体的超声造影剂。造影剂微泡携带基因或药物的方式分两种：黏附法和整合法，其中整合法具有明显的优势，原因在于：黏附法仅仅通过简单的混合将药物或基因黏附在造影剂微泡的表面，一方面结合量少，另一方面黏附的药物或基因与微泡结合不牢固，经静脉注射后，在血流冲击下很容易脱落，靶向性差；而整合法既可以将药物或基因黏附在微泡表面，更可将其整合在微泡膜上或包裹在微泡内，在使结合量增加的同时，又使靶向性更好。通常采用声振法时，在制备过程中产生较高的温度，在制备时不能同时加入药物或基因，只能采用黏附法制备，本发明装置在制备过程中产热少，因此可采用整合法制备兼药物或基因靶向载体的超声造影剂。 Moreover, the device and method of the present invention generate less heat during the preparation of the contrast agent, so they are especially suitable for the preparation of ultrasound contrast agents that are also drug or gene targeting carriers. There are two ways for contrast agent microbubbles to carry genes or drugs: adhesion method and integration method. The integration method has obvious advantages because the adhesion method only adheres drugs or genes to the surface of contrast agent microbubbles through simple mixing. , on the one hand, the amount of binding is small, on the other hand, the adhered drug or gene is not firmly bound to the microbubble, and after intravenous injection, it is easy to fall off under the impact of blood flow, and the targeting is poor; while the integration method can both adhere to the drug or gene On the surface of microbubbles, it can be integrated on the microbubble membrane or wrapped in the microbubbles, which can increase the binding amount and make the targeting better. Usually, when the acoustic vibration method is used, a higher temperature is generated during the preparation process, and drugs or genes cannot be added at the same time during the preparation, and it can only be prepared by the adhesion method. The device of the present invention generates less heat during the preparation process, so it can be prepared by the integration method Ultrasound contrast agent that is also a drug or gene targeting carrier. the

本超声造影剂制备简便、效果好且安全，具有很高的应用价值。同时，如上所述，本发明还可用于制备微液滴。 The ultrasound contrast agent is easy to prepare, effective and safe, and has high application value. Also, as mentioned above, the present invention can also be used to prepare micro-droplets. the

针对一个气体入口、两个液体入口构成的最简单的微泡制备单元，其生产 速率约为107个/分钟，本专利中所述大规模微流控芯片可显著提高产率，如集成128个微泡制备单元，则其产率可提高到109个/分钟，此外，与已有的商品化超声造影剂相比，如美国食品及药物管理局(FDA)批准临床使用的一种超声造影剂Difinity，其产率约为109/45秒，本专利所述大规模微流控芯片的产率可与其相媲美，更重要的是，Difinity造影剂的粒径分布具有多分散性，粒径分布较宽，平均直径在1.8μm左右，直径的标准偏差为1.5μm，但是最大的微泡直径竟达到20μm，从而导致微泡的共振频率范围较大，而现在的超声成像系统的带宽限制只能探测到很窄的共振频率范围，所以大约只有18％的微泡信号可检测到，换言之，82％的微泡不起作用而浪费掉，减少了成像系统的灵敏性，本专利所述装置制备的造影剂具有很高的单分散性，具有有效信号的微泡比例高于92％，可提高超声造影成像技术的灵敏度，特别是在超声分子影像技术中，靶向造影剂的黏附率约为每平方毫米5-25个，微泡的单分散性对提高成像灵敏度显得尤为重要，因此，在这个意义上讲，本专利所述装置制备的造影剂在大规模化生产方面具有绝对的优势，临床应用价值非常高。 For the simplest microbubble preparation unit composed of one gas inlet and two liquid inlets, the production rate is about 107/min. The large-scale microfluidic chip described in this patent can significantly increase the production rate, such as integrating 128 Microbubble preparation unit, its production rate can be increased to 109/min. In addition, compared with the existing commercialized ultrasound contrast agent, such as an ultrasound contrast agent approved by the U.S. Food and Drug Administration (FDA) for clinical use Difinity, its production rate is about 109/45 seconds, the production rate of the large-scale microfluidic chip described in this patent is comparable to it, more importantly, the particle size distribution of Difinity contrast agent has polydispersity, the particle size distribution Wide, the average diameter is about 1.8 μm, and the standard deviation of diameter is 1.5 μm, but the largest microbubble diameter reaches 20 μm, which leads to a large range of resonant frequencies of microbubbles, and the bandwidth limit of the current ultrasound imaging system can only A very narrow range of resonant frequencies is detected, so only about 18% of the microbubble signal can be detected, in other words, 82% of the microbubbles do not work and are wasted, reducing the sensitivity of the imaging system. The device described in this patent is prepared The contrast agent has high monodispersity, and the proportion of microbubbles with effective signal is higher than 92%, which can improve the sensitivity of ultrasound contrast imaging technology, especially in ultrasound molecular imaging technology, the adhesion rate of targeted contrast agent is about 5-25 per square millimeter, the monodispersity of microbubbles is particularly important for improving imaging sensitivity. Therefore, in this sense, the contrast agent prepared by the device described in this patent has an absolute advantage in large-scale production. The clinical application value is very high. the

应当理解的是，对本领域普通技术人员来说，可以根据上述说明加以改进或变换，而所有这些改进和变换都应属于本发明所附权利要求的保护范围。 It should be understood that those skilled in the art can make improvements or changes based on the above description, and all these improvements and changes should belong to the protection scope of the appended claims of the present invention. the

Claims (19)

Translated fromChinese

1.一种集成芯片，其特征在于，以两层相邻结构为一组结构，所述集成芯片包括至少一组结构；1. An integrated chip, characterized in that, with two layers of adjacent structures as a group of structures, the integrated chip includes at least one group of structures;在一组结构中，第一层结构设置M级梯度，第二层结构设置N级梯度；In a group of structures, the first layer structure sets M-level gradients, and the second layer structure sets N-level gradients;其中，M、N是自然数，M小于N；各级梯度中，第一级梯度设置H1个通道1，第二级梯度从每个通道1中引出H2个分支，形成H1×H2个通道2，以此类推，第N级梯度从每个通道(N-1)中引出HN个分支，形成H1×H2......×HN个通道N；Among them, M and N are natural numbers, and M is less than N; among the gradients at all levels, H1 channels 1 are set in the first level gradient, and H2 branches are drawn from each channel 1 in the second level gradient, forming H1×H2 channels 2, By analogy, the N-level gradient leads to HN branches from each channel (N-1), forming H1×H2...×HN channels N;在一组结构中，对于第一层结构的任一特定通道M，均对应于第二层结构从一通道M所引出的在第N级梯度形成H(M+1)×H(M+2)......×H(N-1)×HN数量的特定通道N；In a group of structures, for any specific channel M of the first layer structure, it corresponds to the N-th level gradient formed by the second layer structure drawn from a channel M H(M+1)×H(M+2 )......×H(N-1)×HN number of specific channels N;其中，in,各特定通道N的出口，等距分布在一预设置半径圆形的圆周；The outlets of each specific channel N are equidistantly distributed on the circumference of a circle with a preset radius;对于该特定通道M，其输出方向垂直于该圆形，其出口与该圆形的垂线通过该圆形的圆心，该特定通道M的出口与该圆心距离一特定长度；For the specific channel M, its output direction is perpendicular to the circle, the perpendicular line between the outlet and the circle passes through the center of the circle, and the outlet of the specific channel M is a certain distance from the center of the circle;各特定通道N的输出方向与所述垂线的夹角为相同锐角或直角；The angle between the output direction of each specific channel N and the vertical line is the same acute angle or right angle;沿该特定通道M的输出方向，在该圆形远离该特定通道M的一侧，设置一收集通道；Along the output direction of the specific channel M, a collection channel is provided on the side of the circle away from the specific channel M;所述集成芯片还包括至少一与各收集通道的输出端相连接的收集槽，用于收集和导出产物。The integrated chip also includes at least one collection tank connected to the output end of each collection channel for collecting and discharging products.2.根据权利要求1所述集成芯片，其特征在于，M＝N-1。2. The integrated chip according to claim 1, wherein M=N-1.3.根据权利要求1所述集成芯片，其特征在于，各特定通道N的输出方向通过该圆形圆心。3. The integrated chip according to claim 1, wherein the output direction of each specific channel N passes through the center of the circle.4.根据权利要求1所述集成芯片，其特征在于，通道M和通道N的横截面为矩形；所述矩形高度为20μm至30μm、宽度分别为30μm至50μm、50μm 至100μm，或者，所述通道M和通道N的横截面矩形宽度分别为50μm至100μm、30μm至50μm。4. The integrated chip according to claim 1, wherein the cross-sections of the channel M and the channel N are rectangular; the height of the rectangle is 20 μm to 30 μm, and the width is respectively 30 μm to 50 μm and 50 μm to 100 μm, or, the The cross-sectional rectangular widths of channel M and channel N are 50 μm to 100 μm and 30 μm to 50 μm, respectively.5.根据权利要求1所述集成芯片，其特征在于，所述收集通道的结构至少包括以下结构其中之一：外向放大的锥体、球体、半球体或锥台体，其输入端的管径为7μm至25μm，高度为20μm至30μm；或者，圆柱体或长方体，其沿对应的特定通道M输出方向的截面积略小于该特定通道M沿该方向的截面积，高度为20μm至30μm。5. according to the described integrated chip of claim 1, it is characterized in that, the structure of described collection channel comprises at least one of the following structures: outward enlarged cone, sphere, hemisphere or frustum of cone, the pipe diameter of its input end is 7 μm to 25 μm, and a height of 20 μm to 30 μm; or, a cylinder or a cuboid, whose cross-sectional area along the output direction of a corresponding specific channel M is slightly smaller than the cross-sectional area of the specific channel M along this direction, and a height of 20 μm to 30 μm.6.根据权利要求1所述集成芯片，其特征在于，其基质为表面亲水性基质。6. The integrated chip according to claim 1, wherein the substrate is a surface hydrophilic substrate.7.根据权利要求6所述集成芯片，其特征在于，其基质为硅、玻璃、聚二甲基硅氧烷、聚甲基丙烯酸甲酯或聚碳酯。7. The integrated chip according to claim 6, wherein the substrate is silicon, glass, polydimethylsiloxane, polymethyl methacrylate or polycarbonate.8.根据权利要求1所述集成芯片，其特征在于，第一层结构与第二层结构的组合，为气体通道结构与液体通道结构的组合。8. The integrated chip according to claim 1, wherein the combination of the first layer structure and the second layer structure is a combination of a gas channel structure and a liquid channel structure.9.一种装置，其特征在于，包括：第一物质输入单元、第二物质输入单元、至少一如权利要求1至权利要求8任一所述集成芯片形成集成芯片阵列、贮存单元；9. A device, characterized in that it comprises: a first substance input unit, a second substance input unit, at least one integrated chip as claimed in any one of claims 1 to 8 to form an integrated chip array, and a storage unit;各集成芯片的收集槽，分别与所述贮存单元相连接，用于收集和导出产物；The collection tanks of each integrated chip are respectively connected to the storage unit for collecting and exporting products;所述第一物质输入单元分别与各集成芯片的一层结构的各通道1相连接，用于输入第一物质；The first substance input unit is respectively connected to each channel 1 of the one-layer structure of each integrated chip, and is used for inputting the first substance;所述第二物质输入单元分别从各集成芯片的另一层结构的各通道1相连接，用于输入第二物质。The second substance input unit is respectively connected to each channel 1 of another layer structure of each integrated chip for inputting the second substance.10.根据权利要求9所述装置，其特征在于，所述第一物质输入单元与所述第二物质输入单元的组合，为气体输入单元与液体输入单元的组合；或者，所述第一物质输入单元与所述第二物质输入单元均为液体输入单元。10. The device according to claim 9, wherein the combination of the first substance input unit and the second substance input unit is a combination of a gas input unit and a liquid input unit; or, the first substance Both the input unit and the second substance input unit are liquid input units.11.根据权利要求10所述装置，其特征在于，所述气体输入单元包括顺序连接的压力储气罐、减压阀、第一传输管、微流量计、调压阀和第二传输管，所述第二传输管分别与各集成芯片的一层结构的各通道1相连接；11. The device according to claim 10, wherein the gas input unit comprises a pressure gas storage tank, a pressure reducing valve, a first delivery pipe, a microflow meter, a pressure regulating valve and a second delivery pipe connected in sequence, The second transmission pipe is respectively connected to each channel 1 of the one-layer structure of each integrated chip;所述液体输入单元包括顺序连接的储液器、第三传输管、数字控制式注射泵和第四传输管，所述第四传输管分别与各集成芯片的另一层结构的各通道1相连接。The liquid input unit includes a liquid reservoir, a third transfer tube, a digitally controlled syringe pump and a fourth transfer tube connected in sequence, and the fourth transfer tube is connected to each channel 1 of another layer structure of each integrated chip respectively. connect.12.一种制备微米级分散体的方法，其特征在于，应用于一包括集成芯片阵列的装置中，所述集成芯片阵列由至少一如权利要求1至权利要求8任一所述集成芯片组成；12. A method for preparing a micron-scale dispersion, characterized in that it is applied to a device comprising an array of integrated chips, and the array of integrated chips is composed of at least one integrated chip as described in any one of claims 1 to 8 ;所述方法包括以下步骤：The method comprises the steps of:A1、按第一预设置条件，向各集成芯片的一层结构的各通道1输入第一物质；按第二预设置条件，向各集成芯片的另一层结构的各通道1输入第二物质；A1. According to the first preset condition, input the first substance to each channel 1 of the one-layer structure of each integrated chip; according to the second preset condition, input the second substance to each channel 1 of another layer structure of each integrated chip ;A2、调节第一物质和/或第二物质的输入条件，使各个第一层结构的各通道M输出的物质以同轴流动形式，由对应的各特定通道N的物质按轴向进行包裹， 分别形成待收集产物流，其轴向与所述垂线重合；A2. Adjust the input conditions of the first substance and/or the second substance, so that the substances output by each channel M of each first layer structure are in the form of coaxial flow, and are wrapped axially by the corresponding substances of each specific channel N, Respectively form the product flow to be collected, whose axial direction coincides with the vertical line;A3、从各收集通道的输出端收集和导出产物。A3. Collect and export products from the output ends of each collection channel.13.根据权利要求12所述方法，其特征在于，所述第一物质与所述第二物质的组合，为气体与液体的组合；所述气体至少包括氮气、氟碳类气体、氟硫类气体其中之一；所述液体至少包括磷脂类液体、表面活性剂类液体、上述液体的改性液其中之一。13. The method according to claim 12, characterized in that the combination of the first substance and the second substance is a combination of gas and liquid; the gas at least includes nitrogen, fluorocarbon gases, fluorine sulfur One of the gases; the liquid at least includes one of the phospholipid liquid, the surfactant liquid, and the modified liquid of the above liquids.14.根据权利要求12所述方法，其特征在于，步骤A1中，向各集成芯片的第一层结构的各通道1输入第一液体，向各集成芯片的第二层结构的各通道1输入气体或第二液体；14. The method according to claim 12, characterized in that, in step A1, the first liquid is input to each channel 1 of the first layer structure of each integrated chip, and the first liquid is input to each channel 1 of the second layer structure of each integrated chip. gas or second liquid;步骤A2中，使各个第一层结构的各通道M输出的第一液体以同轴流动形式，由气体或第二液体按轴向包裹所述第一液体，分别形成待收集微液滴流。In step A2, the first liquid output from each channel M of each first layer structure is coaxially flowed, and the gas or the second liquid axially wraps the first liquid to form a micro-liquid droplet flow to be collected respectively.15.根据权利要求12所述方法，其特征在于，步骤A1中，向各集成芯片的一层结构的各通道1输入气体，压力低于10psi，向各集成芯片的另一层结构的各通道1输入液体，流速低于3μL/s；步骤A2中，调节气体和/或液体的输入条件。15. The method according to claim 12, characterized in that, in step A1, each channel 1 of the one-layer structure of each integrated chip is input gas, and the pressure is lower than 10psi, and the gas is injected into each channel of another layer structure of each integrated chip. 1 Input liquid, the flow rate is lower than 3 μL/s; in step A2, adjust the input conditions of gas and/or liquid.16.根据权利要求15所述方法，其特征在于，步骤A1中，输入气体的压力低于5psi，输入液体的流速低于1.5μL/s。16. The method according to claim 15, wherein in step A1, the pressure of the input gas is lower than 5 psi, and the flow rate of the input liquid is lower than 1.5 μL/s.17.根据权利要求15所述方法，其特征在于，步骤A1中，所述液体预先加入特异性配体；或者，步骤A3之后，向所述产物加入特异性配体。17. The method according to claim 15, characterized in that, in step A1, specific ligands are added to the liquid in advance; or, after step A3, specific ligands are added to the product.18.根据权利要求15所述方法，其特征在于，步骤A1中，向各集成芯片的第一层结构的各通道1输入气体，向各集成芯片的第二层结构的各通道1输入液体；18. The method according to claim 15, wherein in step A1, gas is input to each channel 1 of the first layer structure of each integrated chip, and liquid is input to each channel 1 of the second layer structure of each integrated chip;步骤A2中，调节气体和/或液体的输入条件，使各个第一层结构的各通道M输出的气体以同轴流动形式，按轴向包裹液体，分别形成待收集微球流。In step A2, adjust the input conditions of gas and/or liquid, so that the gas output from each channel M of each first layer structure flows coaxially and wraps the liquid axially to form a flow of microspheres to be collected respectively.19.根据权利要求18所述方法，其特征在于，步骤A2中，各个第一层结构的各通道M输出的气体，分别在两侧高速流动的液体聚焦作用下形成中轴线位于所述垂线位置的稳定倒锥体，各倒锥体的尖端分别与各收集通道位置相对。 19. The method according to claim 18, characterized in that, in step A2, the gas output from each channel M of each first layer structure is formed under the focusing action of liquids flowing at high speed on both sides to form a central axis located on the vertical line The position of the stable inverted cone, the tip of each inverted cone is respectively opposite to the position of each collection channel. the

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