微流控芯片酶分析及其在酶抑制剂筛选中的应用

微流控芯片以其消耗少、易于微型化和集成化等优点在酶分析领域占有重要地位。近年来随着新检测技术的不断出现,酶抑制剂筛选芯片的结构也从简单的"混合-反应"和"分离-检测"变得更加多样化和多功能化。微流控芯片上分子固定化酶、细胞培养等技术的进步为微流控芯片上实现酶抑制剂的高通量和高内涵筛选带来了巨大优势。本文对用于酶分析的微流控芯片的种类和构型进行简介和归纳总结,重点讨论和综述了其在酶抑制剂筛选中的应用及其最新研究进展。     

微流控芯片的研究及产业化

以大连研究团队的近期工作为基础,结合2015年末召开的“深圳-大连微流控芯片及其产业化战略研讨会”内容,扼要阐述作者对近期微流控芯片的研究及产业化的基本看法.鉴于微流控芯片研究的主流已从平台构建和方法发展转为不同领域的广泛应用,本文重点介绍了微流控芯片在现代生物化学分析、即时诊断、材料筛选-材料合成以及组织-器官仿生等4个应用领域的研究趋势,讨论了3D打印技术的崛起对微流控芯片的影响和挑战,阐述了微流控芯片作为当代极为重要的新兴科学技术平台和国家层面产业转型的潜在战略领域,在全球范围内产业化的发展势头.全文引用文献69篇.     

微流控技术筛选酶抑制剂的研究进展

随着近年新合成或提取的化合物大量涌现,药物筛选朝着快速、高效、高通量方向发展。微流控分析技术具有的分析微型化、高通量化、可集成化和良好的生物相容性等特点,为药物的筛选提供了新的方法和技术平台。本文简要介绍了酶抑制剂筛选,重点评述基于微流控技术筛选酶抑制剂的研究进展。     

微流控芯片荧光检测系统的研究进展北大核心CSCD

微流控芯片又称芯片实验室,具有检测高效、消耗试剂少、高通量、微型化和集成化等特点,许多检测方式(如光学检测、电化学检测)已经集成于微流控芯片上,而荧光检测是微流控芯片检测技术的常见手段之一。为此,在介绍了荧光检测技术的基本原理和光路结构的基础上,从激发光源、光传辅助手段和检测器等方面综述了微流控芯片荧光检测系统的研究进展,并对其发展进行了展望(引用文献55篇)。     

微流控分析芯片上生化反应技术研究进展*

综述了近几年来在微流控芯片上以检测或动力学研究为目的而进行的各种生物化学反应技术,包括酶反应、免疫反应、酶联免疫反应、PCR反应,含氮氧化物释放反应。对非均相反应的蛋白质在微管道中的固定技术,均相反应的管道设计技术,以及在微流控芯片上集成生化反应的优越性等问题做了详细的讨论。     

3D打印微流控芯片技术研究进展

近年来,微流控技术在生命科学和医学诊断等领域得到广泛的应用,显示出了其在检测速度、精度以及试剂损耗等方面相比传统方法的显著优势.然而,使用从半导体加工技术继承而来的微加工技术制作微流控芯片具有比较高的资金和技术门槛,在一定程度上阻碍了微流控技术的推广和应用.近年来随着3D打印技术的兴起,越来越多的研究者尝试使用3D打印技术加工微流控芯片.相比于传统的微加工技术,3D打印微流控芯片技术显示出了其设计加工快速、材料适应性广、成本低廉等优势.本文针对近年来国内外在3D打印微流控芯片领域的最新进展进行了综述,着重介绍了采用微立体光刻、熔融沉积成型以及喷墨打印等3D打印技术加工制作微流控芯片的方法,以及这些微流控芯片在分析化学、生命科学、医学诊断等领域的应用,并对3D打印微流控芯片技术未来的发展进行了展望.     

微流控SERS芯片的设计制备及其在细菌检测中的应用

微流控芯片分析平台与表面增强拉曼散射(Surface enhanced Raman scattering,SERS)光谱分析方法结合,充分利用了SERS法所具备的样品前处理简便、检测无损、成分辨识度高以及适宜水环境检测等优点,在生化分析检测领域备受关注。微流控SERS芯片设计及芯片上SERS增强基质的制备是构建微流控SERS芯片分析方法和系统的关键,也是提高检测灵敏度和可重复性的核心问题。该文在介绍微流控SERS芯片的基本构型和功能的基础上,重点综述了微流控SERS芯片上SERS基质的制备方法及其测试效果。基于微电子机械系统(Micro-Electro-mechanical-System,MEMS)加工技术制备的SERS基质,具有纳米粒径有序可控、便于集成制备但增强基质材料种类有限的特点;基于化学沉积和自组装等理化方法制备的SERS基质具有基质种类易拓展、成本低、与微流控通道结合方法灵活等特点。在这些基础上构建的微流控SERS芯片及其分析测试方法和系统,在细菌等许多生化检测领域显示出强大的发展潜力。     

微流控芯片上电驱动在线富集技术的研究进展

微流控芯片上电驱动在线富集技术是一种有效提高分析效率、检测灵敏度和降低对检测器要求的技术和方法。本文针对目前微流控芯片分析系统中生化样品的预处理问题,对芯片上电驱动在线富集技术进行了分析讨论,介绍了等速电泳、等电聚焦、场放大和介电电泳的样品预富集技术在微流控芯片上的实现与应用,并对每一种技术的原理、特点、存在的问题、近年发展的状况和发展趋势进行了综述。     

近期微流控芯片疾病诊断技术的研究进展

微流控芯片具有液流可控、样品消耗量小、反应速度快、易于集成化等特点,在临床诊断和疾病筛查领域具有广阔的发展前景。本文针对近年来微流控芯片技术在疾病诊断方面的最新研究进展,从疾病标志物检测、细胞筛选和药物代谢研究及疾病诊断微流控芯片装置的发展现状等方面概述其在疾病诊断方面的应用和发展。     

微流控芯片上的免疫分析

近20年来,随着微流控芯片加工技术的不断发展,微流控分析已从一个概念发展为当前世界上最前沿的科技领域之一,微流控芯片上免疫分析的方法研究也取得重要进展。这些芯片包含传输流体的微通道和免疫分析程序中部分或全部的必要组件。微流控技术用于免疫分析在减少试剂用量、缩短分析时间、自动化等方面提高了分析性能。本文综述了微流控芯片上免疫分析的发展、分类,并评述了各类微流控免疫分析芯片的性能及优缺点。     

Solid supports for micro analytical systems

The development of micro analytical systems requires that fluids are able to interact with the surface of the microfluidic chip in order to perform analysis such as chromatography, solid phase extraction, and enzymatic digestion. These types of analyses are more efficient if there are solid supports within the microfluidic channels. In addition, solid supports within microfluidic chips are useful in producing devices with multiple functionalities. In recent years there have been many approaches introduced for incorporating solid supports within chips. This review will explore several state of the art methods and applications of introducing solid supports into chips. These include packing chips with beads, incorporating membranes into chips, creating supports using microfabrication, and fabricating gels and polymer monoliths within microfluidic channels.     

微流控纸芯片在环境分析检测中的应用

近年来,微流控纸芯片由于低成本、便携化、检测快等优点,在需要快速检测的环境分析领域中展现出了巨大的应用前景。该综述从微流控纸芯片在环境分析中的应用角度,总结归纳了微流控纸芯片在环境分析中的最新研究进展,并展望了其在未来的发展趋势与挑战。论文内容引用150余篇源于科学引文索引(SCI)与中文核心期刊中的相关论文。该综述包括微流控纸芯片在环境检测中的优势与制造方法介绍;电化学法、荧光法、比色法、表面增强拉曼法、集成传感法等基于纸芯片的先进分析方法介绍;根据环境分析目标物种类,如重金属离子、营养盐、农药、微生物、抗生素以及其他污染物等,对纸芯片的最新应用现状进行了举例评述;基于微流控纸芯片的环境分析研究的未来发展趋势和前景展望。通过综述近期相关研究,表明微流控纸芯片从提出至今虽然只有十几年的发展历程,但其在环境分析研究中的发展却十分迅速。微流控纸芯片可以根据不同的环境条件和检测要求灵活选择制作与分析方法,实现最佳的检测效果。但是微流控纸芯片也面临一些挑战,如纸张机械强度不足、流体控制程度不佳等问题。这些问题指出了微流控纸芯片在环境检测领域的发展趋势,相信随着不断深入的研究,纸芯片将会在未来的环境分析中发挥更大作用。     

Rapid fabrication of electrochemical enzyme sensor chip using polydimethylsiloxane microfluidic channel

Applicability of polydimethylsiloxane (PDMS) for easy and rapid fabrication of enzyme sensor chips, based on electrochemical detection, is examined. The sensor chip consists of PDMS substrate with a microfluidic channel fabricated in it, and a glass substrate with enzyme-modified microelectrodes. The two substrates are clamped together between plastic plates. The sensor chip has shown no leakage around the microelectrodes under continuous solution flow (34 μl/min). Amperometric response of the sensor chips developed in this work suggest that various types of enzyme sensors can be designed by using PDMS microfluidic channels.     

Biochemical analysis with microfluidic systems

Microfluidic systems are capillary networks of varying complexity fabricated originally in silicon, but nowadays in glass and polymeric substrates. Flow of liquid is mainly controlled by use of electroosmotic effects, i.e. application of electric fields, in addition to pressurized flow, i.e. application of pressure or vacuum. Because electroosmotic flow rates depend on the charge densities on the walls of capillaries, they are influenced by substrate material, fabrication processes, surface pretreatment procedures, and buffer additives. Microfluidic systems combine the properties of capillary electrophoretic systems and flow-through analytical systems, and thus biochemical analytical assays have been developed utilizing and integrating both aspects. Proteins, peptides, and nucleic acids can be separated because of their different electrophoretic mobility; detection is achieved with fluorescence detectors. For protein analysis, in particular, interfaces between microfluidic chips and mass spectrometers were developed. Further levels of integration of required sample-treatment steps were achieved by integration of protein digestion by immobilized trypsin and amplification of nucleic acids by the polymerase chain reaction. Kinetic constants of enzyme reactions were determined by adjusting different degrees of dilution of enzyme substrates or inhibitors within a single chip utilizing mainly the properties of controlled dosing and mixing liquids within a chip. For analysis of kinase reactions, however, a combination of a reaction step (enzyme with substrate and inhibitor) and a separation step (enzyme substrate and reaction product) was required. Microfluidic chips also enable separation of analytes from sample matrix constituents, which can interfere with quantitative determination, if they have different electrophoretic mobilities. In addition to analysis of nucleic acids and enzymes, immunoassays are the third group of analytical assays performed in microfluidic chips. They utilize either affinity capillary electrophoresis as a homogeneous assay format, or immobilized antigens or antibodies in heterogeneous assays with serial supply of reagents and washing solutions.     

Chiral separations on multichannel microfluidic chips

Chiral separations of FITC-labeled basic drugs on multichannel microfluidic chips with LIF detector were investigated. A preliminary screening procedure for seven neutral CDs was performed under optimized conditions for chiral separations of three FITC-labeled drugs (baclofen, norfenefrine, and tocainide) on a mono-channel microfluidic chip. According to the results of screening, FITC-baclofen and FITC-norfenefrine as well as two chiral selectors including gamma-CD and dimethyl-beta-CD (DM-beta-CD) were selected as models to perform chiral separations on a two-channel chip. FITC-baclofen enantiomers were separated completely by gamma-CD in one channel, while resolution of FITC-norfenefrine enantiomers was achieved by DM-beta-CD in the other channel in the same run. Furthermore, the feasibility of using one chiral selector to separate multiple chiral samples was studied on a four-channel chip. These results show that multichannel chip has a potential for chiral high-throughput screening.     

Analytical and numerical study of Joule heating effects on electrokinetically pumped continuous flow PCR chips

Joule heating is an inevitable phenomenon for microfluidic chips involving electrokinetic pumping, and it becomes a more important issue when chips are made of polymeric materials because of their low thermal conductivities. Therefore, it is very important to develop methods for evaluating Joule heating effects in microfluidic chips in a relatively easy manner. To this end, two analytical models have been established and solved using the Green's function for evaluating Joule heating effects on the temperature distribution in a microfluidic-based PCR chip. The first simplified model focuses on the understanding of Joule heating effects by ignoring the influences of the boundary conditions. The second model aims to consider practical experimental conditions. The analytical solutions to the two models are particularly useful in providing guidance for microfluidic chip design and operation prior to expensive chip fabrication and characterization. To validate the analytical solutions, a 3-D numerical model has also been developed and the simultaneous solution to this model allows the temperature distribution in a microfluidic PCR chip to be obtained, which is used to compare with the analytical results. The developed numerical model has been applied for parametric studies of Joule heating effects on the temperature control of microfluidic chips.     

Advances in the preparation of porous polymer monoliths in capillaries and microfluidic chips with focus on morphological aspects

Porous polymer monoliths have emerged as unique materials for many applications, including liquid-chromatographic analyses at an unrivaled speed, solid-phase extraction, and enzyme immobilization in capillary and microfluidic chip format. This article reviews the state of the art in the preparation of monoliths in narrow-bore capillaries and microfluidic chips and their miniaturization under conditions of spatial confinement. New developments in their preparation mainly using free radical polymerization techniques with a focus on morphological aspects in view of homogeneous porous materials are described. The suitability of monoliths for analysis of both large and small molecules is also discussed.     

基于微流控芯片的乳腺癌细胞微环境酸化检测

建立了基于微流控芯片的乳腺癌微环境酸化模型和动态检测微环境酸化情况的分析方法。设计了一种多层复合式微流控芯片,将乳腺癌细胞悬液引入含有水凝胶前体的芯片培养室后,在硝酸纤维素薄膜上固化形成3D培养支架。芯片通道连续灌流模拟血流供应,并将非电化学的pH检测器引入芯片,通过图像分析得到实时的pH变化。通过观察癌细胞的存活率、增殖率、乳酸水平及pH值,分析微环境的酸化情况,同时与正常细胞进行比较。结果表明,连续灌流培养7 d,乳腺癌细胞的存活率保持在90%以上;随着培养天数的增加,芯片上癌细胞微环境的pH值逐渐降低,且灌流速度越低,pH值下降越明显,而正常细胞微环境的pH值无明显变化。基于微流控芯片的微环境酸化检测平台可实时动态检测微环境的pH值,有望成为相关肿瘤研究的有力工具。     

DNA Nanolithography Enables a Highly Ordered Recognition Interface in a Microfluidic Chip for the Efficient Capture and Release of Circulating Tumor Cells

Microfluidic chips with nano‐scale structures have shown great potential, but the fabrication and cost issues restrict their application. Herein, we propose a conceptually new “DNA nanolithography in a microfluidic chip” by using sub‐10 nm three‐dimensional DNA structures (TDNs) as frameworks with a pendant aptamer at the top vertex (ApTDN‐Chip). The nano‐scale framework ensures that the aptamer is in a highly ordered upright orientation, avoiding the undesired orientation or crowding effects caused by conventional microfluidic interface fabrication processes. Compared with a monovalent aptamer modified chip, the capture efficiency of ApTDN‐Chip was enhanced nearly 60 % due to the highly precise dimension and rigid framework of TDNs. In addition, the scaffolds make DNase I more accessible to the aptamer with up to 83 % release efficiency and 91 % cell viability, which is fully compatible with downstream molecular analysis. Overall, this strategy provides a novel perspective on engineering nano‐scaffolds to achieve a more ordered nano‐topography of microfluidic chips.     

Parameters Governing the Formation of Photopolymerized Silica Sol–Gel Monoliths in PDMS Microfluidic Chips

Although polydimethylsiloxane (PDMS) microfluidic chips provide an alternative to more expensive microfabricated glass chips, formation of monolithic stationary phases in PDMS is not a trivial task. Photopolymerized silica sol–gel monoliths were fabricated in PDMS-based microfluidic devices using 3-trimethoxysilylpropylmethacrylate and glycidyloxypropyltrimethoxysilane. The monolith formation was optimized by identifying a suitable porogen, controlling monomer concentration, functional additives, salts, porogen, wall attachment methods, and rinsing procedures. The resulting monoliths were evaluated using scanning electron microscopy, image analysis, differential scanning calorimetry, and separation performance. Monoliths functionalized with boronic acid ligands were used for the separation of cis-diol containing compounds both in batch mode and in the microfluidic chip.