微流控芯片单细胞进样和溶膜

单细胞分析对重大疾病的早期诊断、治疗和药物筛选以及细胞生理、病理过程的研究有重要意义.将毛细管电泳用于单细胞多组分的测定已取得一些成果,但受毛细管的一维结构限制,单细胞进样和溶膜操作较复杂.微流控分析芯片的网络结构和微米级的通道尺寸使简化单细胞分析成为可能.     

微流控芯片系统在单细胞研究中的应用

微流控芯片具有网络式通道结构,扩展了在细胞和亚细胞水平进行生命科学研究的能力,为单细胞研究提供了一个新的平台.在微流控芯片通道中,人们利用气压、液压和电压,或利用介电电泳、光学陷阱、行波介电电泳以及磁场等技术,可以操纵细胞通过或驻留在通道内的任意位置,从而使单细胞计数、筛选以及胞内组分分析等操作大大简化.本文对微流控芯片系统在血液流变学、单细胞操纵与计数以及单细胞胞内组分分析中的应用进行了综述,介绍了用于单细胞研究的多种微芯片系统,讨论了芯片上进行单细胞操纵的各种方法     

微流控芯片毛细管电泳在蛋白质分离分析中的应用研究进展

重点介绍了近年来国内外在微流控芯片毛细管电泳法用于蛋白质分离分析方面的研究进展。按照分离模式的不同,综述了各种应用于蛋白质分离的微流控芯片毛细管电泳系统,讨论了抑制芯片中的蛋白吸附的各种方法,并展望了芯片毛细管电泳系统在蛋白质分离领域的发展前景。引用文献47篇。     

微流控芯片操纵传输及实时监测单细胞量子释放

微流控芯片技术用于细胞生化分析已引起了广泛关注.Harrison等首次在微流控芯片上对细胞群体进行操纵、传输及反应.yang等在微流控芯片上操纵细胞群体的排列,并用荧光检测细胞群体摄取钙的反应.至今还未见到微流控芯片对单个细胞进行操纵传输、定位及实时监测的报道.单细胞受激释放的监测对探索生物体神经传导具有重要意义.     

蛋白质/多肽的微流控二维芯片电泳*

在微流控芯片上构建多维分离系统,为蛋白质组学研究提供了一个有发展前景的高效分离分析技术平台。本文介绍了二维芯片电泳系统耦联模式选取及正交性评价的方法;综述了针对蛋白质/多肽分离分析的各种耦联模式微流控二维芯片电泳分析系统,如胶束电动力学色谱（MEKC）与毛细管区带电泳（CZE）,开管电色谱（OECE）与CZE,等电聚焦（IEF）与CZE, IEF与SDS毛细管凝胶电泳（CGE）, SDS-CGE与MEKC等。特别对二维电泳芯片切换接口的类型进行了分类,探讨了用于微流控二维芯片电泳系统的检测技术,并展望了微流控二维电泳芯片在蛋白质组学研究中的应用前景和发展方向。     

毛细管电泳和微流控芯片技术在临床尿检中的应用

本文就毛细管电泳和微流控芯片技术在临床尿检中的应用,以及毛细管电泳和微流控芯片技术在尿样前处理、样品富集方面的进展进行了综述。主要介绍了临床尿液一般化学检查和特殊化学检查,着重对肾功能指标的生化检查进行了总结。根据目前的研究状况,对毛细管电泳和微流控芯片技术在临床检测上的应用前景和发展方向进行了展望。     

微流控芯片单细胞进样,溶膜及分离检测胞内谷胱甘肽

单细胞分析对重大疾病的早期诊断等方面有重要意义[1].微流控分析芯片的网络结构和微米级的通道尺寸适合于单细胞进样、溶膜和分离分析.但目前的报道主要集中在细胞培养、计数和筛选[2].我们在十字通道微流控芯片上,通过调节储液池的液面高度和细胞悬液密度,使单细胞逐个通过芯片进样通道和分离通道之间的区域,再结合控制电渗流方向,使单细胞固定在分离通指定位置,然后用电泳缓冲液结合高电场实现细胞快速溶膜,接着进行电泳分离和LIF检测.实现了单个血红细胞内谷胱甘肽(GSH)的高效分离及定量分析.     

基于毛细管电泳及微流控芯片的药物提取分离技术

近年来,在提取分离方面出现了许多新技术和新方法.其中毛细管电泳和微流控芯片技术以其微量、高效、快速等特点,在药物提取分离中已渐显优势.该文对基于毛细管电泳和微流控芯片的两相电泳技术、微流控液液萃取技术、微流控固液萃取技术、微流控过滤式分离技术、微流控膜分离技术在药物分离提取中的应用进行了综述.     

基于流体动力单细胞高效捕获的微流控芯片结构研究进展

单细胞分析对于重大疾病的早期诊断及治疗、药物筛选和生理病理过程的研究具有重要意义。微流控芯片能够精确控制单细胞的微环境,实时监测单细胞的行为,已成为单细胞分析的强大工具。单细胞捕获是单细胞分析的重要步骤。目前已报道了多种微流控芯片用于单细胞捕获的方法,其中基于流体动力的微流控芯片单细胞捕获方法具有操作方便、单细胞捕获效率高等优点,受到研究人员的广泛关注及使用。为了全面了解基于流体动力的微流控芯片单细胞捕获方法的研究现状,掌握单细胞高效捕获的微流控芯片结构设计,实现单细胞精准快速分析,本文综述了基于流体动力的单细胞高效捕获（>70%）原理及微流控芯片结构,根据结构设计不同分为微井结构、微柱结构和旁路通道结构,介绍了单细胞高效捕获的微流控芯片优化过程,总结了微流控芯片的材质、结构特点及单细胞捕获效率等,对不同单细胞捕获结构的优势及不足进行了分析。最后,对基于流体动力的微流控芯片单细胞捕获方法的发展趋势进行了展望。     

基于微流控技术的单细胞生物物理特性表征

单细胞水平的生物物理特性表征,可有效阐明细胞的功能和状态,揭示细胞的单体差异性,对于细胞的分化和病理研究,以及疾病的早期临床诊断和治疗具有非常重要的意义。由于具有与细胞尺度相匹配的微米级腔道,微流控芯片比传统生化方法更适合单细胞样本的微环境精确控制、高通量定向操纵及多参数非特异性检测,已成为单细胞表征与分析的一项重要技术平台。本文总结了基于微流控技术的单细胞生物物理特性表征方法及其应用的最新进展,着重分析微流控芯片在常规方法难以达成的单细胞和高通量研究中的独特优势,最后探讨了微流控单细胞生物物理特性检测芯片在临床应用中面临的挑战和未来的发展动向,并提出一种新型的单细胞多参数同时表征的微流控分析器件。     

Microfluidic chip electrophoresis for biochemical analysis

Microfluidic chip electrophoresis has been widely employed for separation of various biochemical species owing to its advantages of low sample consumption, low cost, fast analysis, high throughput, and integration capability. In this article, we reviewed the development of four different modes of microfluidics‐based electrophoresis technologies including capillary electrophoresis, gel electrophoresis, dielectrophoresis, and field (electric) flow fractionation. Coupling detection schemes on microfluidic electrophoresis platform were also reviewed such as optical, electrochemical, and mass spectrometry method. We further discussed the innovative applications of microfluidic electrophoresis for biomacromolecules (nucleic acids and proteins), biochemical small molecules (amino acids, metabolites, ions, etc.), and bioparticles (cells and pathogens) analysis. The future direction of microfluidic chip electrophoresis was predicted.     

A capillary electrophoresis chip with hydrodynamic sample injection for measurements from a continuous sample flow

A microchip-based capillary electrophoresis device supported by a microfluidic network made of poly(dimethylsiloxane), used for measuring target analytes from a continuous sample flow, is presented. The microsystem was fabricated by means of replica molding in combination with standard microfabrication technologies, resulting in microfluidic components and an electrochemical detector. A new hydrodynamic sample injection procedure is introduced, and the maximum number of consecutive measurements that can be made with a poly(dimethylsiloxane) capillary electrophoresis chip with amperometric detection is investigated with respect to reproducibility. The device features a high degree of functional integration, so the benefits associated with miniaturized analysis systems apply to it.     

Analytical detection techniques for droplet microfluidics—A review

In the last decade, droplet-based microfluidics has undergone rapid progress in the fields of single-cell analysis, digital PCR, protein crystallization and high throughput screening. It has been proved to be a promising platform for performing chemical and biological experiments with ultra-small volumes (picoliter to nanoliter) and ultra-high throughput. The ability to analyze the content in droplet qualitatively and quantitatively is playing an increasing role in the development and application of droplet-based microfluidic systems. In this review, we summarized the analytical detection techniques used in droplet systems and discussed the advantage and disadvantage of each technique through its application. The analytical techniques mentioned in this paper include bright-field microscopy, fluorescence microscopy, laser induced fluorescence, Raman spectroscopy, electrochemistry, capillary electrophoresis, mass spectrometry, nuclear magnetic resonance spectroscopy, absorption detection, chemiluminescence, and sample pretreatment techniques. The importance of analytical detection techniques in enabling new applications is highlighted. We also discuss the future development direction of analytical detection techniques for droplet-based microfluidic systems.     

Development of an ultra-low volume flow cell for surface plasmon resonance detection in a miniaturized capillary electrophoresis system

A miniaturized capillary electrophoresis system coupled to a surface plasmon resonance (SPR) sensor on a microfluidic platform fabricated from PDMS is detailed. A previously described split-flow injection technique is first utilized to manipulate sample into the microfluidic chip, followed by separation within the fused-silica capillary and final off-capillary detection of analytes via SPR. Instead of using commercial SPR flow cells requiring relatively large detection volumes, samples of less than 1 nL volume are utilized. The interface between the CE system and SPR sensor made it possible to detect minute volumes of sample with minimal dispersion. The flow cell has the potential to be applicable to miniaturized flow-injection (FI) systems where submicroliter volumes of sample are frequently only available for analysis. The components present in solution, but not bound to the sensor surface, were also investigated. The sensitivity of the CE-SPR system was similar to that found in UV-spectrometric instruments and nonchromophoric components could also be measured.     

Sampling techniques for single-cell electrophoresis

Cells are extraordinarily complex, containing thousands of different analytes with concentrations spanning at least nine orders of magnitude. Analyzing single cells instead of tissue homogenates provides unique insights into cell-to-cell heterogeneity and aids in distinguishing normal cells from pathological ones. The high sensitivity and low sample consumption of capillary and on-chip electrophoresis, when integrated with fluorescence, electrochemical, and mass spectrometric detection methods, offer an ideal toolset for examining single cells and even subcellular organelles; however, the isolation and loading of such small samples into these devices is challenging. Recent advances have addressed this issue by interfacing a variety of enhanced mechanical, microfluidic, and optical sampling techniques to capillary and on-chip electrophoresis instruments for single-cell analyses.     

Automated analysis of single stem cells in microfluidic traps

We report a reliable strategy to perform automated image cytometry of single (non-adherent) stem cells captured in microfluidic traps. The method rapidly segments images of an entire microfluidic chip based on the detection of horizontal edges of microfluidic channels, from where the position of the trapped cells can be derived and the trapped cells identified with very high precision (>97%). We used this method to successfully quantify the efficiency and spatial distribution of single-cell loading of a microfluidic chip comprised of 2048 single-cell traps. Furthermore, cytometric analysis of trapped primary hematopoietic stem cells (HSC) faithfully recapitulated the distribution of cells in the G1 and S/G2-M phase of the cell cycle that was measured by flow cytometry. This approach should be applicable to automatically track single live cells in a wealth of microfluidic systems.     

Laminar flow mediated continuous single-cell analysis on a novel poly(dimethylsiloxane) microfluidic chip

A novel microfluidic chip with simple design, easy fabrication and low cost, coupled with high-sensitive laser induced fluorescence detection, was developed to provide continuous single-cell analysis based on dynamic cell manipulation in flowing streams. Making use of laminar flows, which formed in microchannels, single cells were aligned and continuously introduced into the sample channel and then detection channel in the chip. In order to rapidly lyse the moving cells and completely transport cellular contents into the detection channel, the angle of the side-flow channels, the asymmetric design of the channels, and the number, shape and layout of micro-obstacles were optimized for effectively redistributing and mixing the laminar flows of single cells suspension, cell lysing reagent and detection buffer. The optimized microfluidic chip was an asymmetric structure of three microchannels, with three microcylinders at the proper positions in the intersections of channels. The microchip was evaluated by detection of anticancer drug doxorubicin (DOX) uptake and membrane surface P-glycoprotein (P-gp) expression in single leukemia K562 cells. An average throughput of 6–8 cells min⁻¹ was achieved. The detection results showed the cellular heterogeneity in DOX uptake and surface P-gp expression within K562 cells. Our researches demonstrated the feasibility and simplicity of the newly developed microfluidic chip for chemical single-cell analysis.     

Quantitative determination of dopamine in single rat pheochromocytoma cells by microchip electrophoresis with only one high‐voltage power supply

We developed a method for the direct identification of dopamine in single cultured rat pheochromocytoma cells by capillary electrophoresis using an end‐channel carbon fiber nanoelectrode amperometric detector. The operation mode was designed to achieve single‐cell injection and lysis in microfluidic chip electrophoresis with only one high‐voltage power supply. The separation and detection conditions were optimized. Four catecholamines were baseline‐separated and determined with this system, and the cell density and liquid height of the reservoirs were accommodated for single cell loading, docking and analysis. The microchip capillary electrophoresis system was successfully applied to determine dopamine in single cultured rat pheochromocytoma cells.     

Rapid Analysis of Oligonucleotides on a Planar Microfluidic Chip

State-of-the-art microfluidic analytical systems are briefly surveyed. Attention is focused on the use of microchip capillary electrophoresis. The main results obtained in the development of a prototype analytical system with a laser-induced fluorescence detector for electrophoresis on a glass microfluidic chip are presented. Experimental data on electroosmotic flow and the distribution of sample fluorescence intensity over the cross section of a microchannel are analyzed. A procedure for the rapid analysis of oligonucleotides on a microfluidic chip is described.