Microfluidic device embedding electrodes for dielectrophoretic manipulation of cells‐A review

Microfluidic device embedding electrodes realizes cell manipulation with the help of dielectrophoresis. Cell manipulation is an important technology for cell sorting and cell population purification. Till now, the theory of dielectrophoresis has been greatly developed. Microfluidic devices with various arrangements of electrodes have been reported from the beginning of the single non‐uniform electric field to the later multiple physical fields. This paper reviews the research status of microfluidic device embedding electrodes for cell manipulation based on dielectrophoresis. Firstly, the working principle of dielectrophoresis is explained. Next, cell manipulation approaches based on dielectrophoresis are introduced. Then, different types of electrode arrangements in the microfluidic device for cell manipulation are discussed, including planar, multilayered and microarray dot electrodes. Finally, the future development trend of the dielectrophoresis with the help of microfluidic devices is prospected. With the rapid development of microfluidic technology, in the near future, high precision, high throughput, high efficiency, multifunctional, portable, economical and practical microfluidic dielectrophoresis will be widely used in the fields of biology, medicine, agriculture and so on.     

Analysis Laboratory on a Chip

Microfluidic chip offers a promising platform for chemical or biological analysis on the basis of flexible integration of various functional operation units. This article provides an overview of the recent achievements of microfluidic chip and its applications based on the works mainly carried out in the authors' lab, especially for the purpose of constructing analytical laboratory on a microfluidic chip. Different operation units and some representative applications in molecules, cell and organism analysis are described.     

微流控芯片测定单细胞内化学组分的进展

细胞是生命的基本单元。由于细胞的个体差异,传统分析群体细胞的方法难以得到单细胞的重要信息。准确可靠地测定单细胞内化学组分的含量能大大提高从正常细胞中辨别不正常细胞的能力,为进一步研究和发展生物化学、医学和临床检验等领域奠定基础。近年来,用微流控芯片进行单细胞分析已引起广泛的兴趣。微流控芯片可以集成单细胞进样、溶膜、电泳分离胞内化学组分和高灵敏度测定等一系列操作步骤,为分析单细胞内的化学组分提供了新的技术平台。本文主要综述了近年来微流控芯片测定单细胞内化学组分的进展。重点在于利用电渗流、压力结合电渗流和激光镊子等技术操控单细胞在微流控芯片上完成单细胞进样、溶膜、细胞内化学组分的电泳分离和高灵敏度测定等一系列操作步骤。对在微流控芯片上的衍生技术也做了较为详细的阐述。     

微流控芯片上细胞培养与分析方法研究进展

微流控芯片以其强大的微流体和微小物质控制能力成为研究单细胞、细胞群落乃至生物组织的重要手段。在本篇综述中,我们将以微流控芯片上细胞体外培养模型的建立为主,对近几年来重要的研究工作加以评述,全面地介绍微流控技术在细胞生命科学研究中应用的优势和未来发展方向,具体包括微流控芯片的细胞操控能力、细胞培养微环境的构建以及芯片联用检测手段,希望为从事这一领域研究工作的读者提供一些新的思路。     

Recent advances in electric analysis of cells in microfluidic systems

Microfluidics offers an ideal platform to integrate cell-based assays with electric measurements. The technological advances in microfluidics, microelectronics, electrochemistry, and electrophysiology have greatly inspired the development of microfluidic/electric devices that work with a low number of cells or single cells. The applications of these microfluidic systems range from the detecting of cell culture density to the probing of cellular functions at the single-cell level. In this review, we introduce the recent advances in the electric analysis of cells on a microfluidic platform, specifically related to the quantification and monitoring of cells in static solution, on-chip patch-clamp measurement, and examination of flowing cells. We also point out future directions and challenges in this field. Figure Different microfluidic devices applied to electrical analysis of cells     

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.     

Biofunctionalization of electrowetting-on-dielectric digital microfluidic chips for miniaturized cell-based applications

In this paper we report on the controlled biofunctionalization of the hydrophobic layer of electrowetting-on-dielectric (EWOD) based microfluidic chips with the aim to execute (adherent) cell-based assays. The biofunctionalization technique involves a dry lift-off method with an easy to remove Parylene-C mask and allows the creation of spatially controlled micropatches of biomolecules in the Teflon-AF(?) layer of the chip. Compared to conventional methods, this method (i) is fully biocompatible; and (ii) leaves the hydrophobicity of the chip surface unaffected by the fabrication process, which is a crucial feature for digital microfluidic chips. In addition, full control of the geometry and the dimensions of the micropatches is achieved, allowing cells to be arrayed as cell clusters or as single cells on the digital microfluidic chip surface. The dry Parylene-C lift-off technique proves to have great potential for precise biofunctionalization of digital microfluidic chips, and can enhance their use for heterogeneous bio-assays that are of interest in various biomedical applications.     

A valve‐based microfluidic device for on‐chip single cell treatments

Assays toward single‐cell analysis have attracted the attention in biological and biomedical researches to reveal cellular mechanisms as well as heterogeneity. Yet nowadays microfluidic devices for single‐cell analysis have several drawbacks: some would cause cell damage due to the hydraulic forces directly acting on cells, while others could not implement biological assays since they could not immobilize cells while manipulating the reagents at the same time. In this work, we presented a two‐layer pneumatic valve‐based platform to implement cell immobilization and treatment on‐chip simultaneously, and cells after treatment could be collected non‐destructively for further analysis. Target cells could be encapsulated in sodium alginate droplets which solidified into hydrogel when reacted with Ca²⁺. The size of hydrogel beads could be precisely controlled by modulating flow rates of continuous/disperse phases. While regulating fluid resistance between the main channel and passages by the integrated pneumatic valves, on‐chip capture and release of hydrogel beads was implemented. As a proof of concept for on‐chip single‐cell treatments, we showed cellular live/dead staining based on our devices. This method would have potential in single cell manipulation for biochemical cellular assays.     

Microfluidic methods for cell separation and subsequent analysis

Cell is the most basic unit of the morphological structure and life activity of an organism. Learning the composition, structure and function of cells, exploring the life activities of cells and studying the interaction between cells are of great significance for human cognition and control of the life activities of organisms. Therefore, rapid, convenient, inexpensive, high-precision and reliable methods of cell separation and analysis are being developed to obtain accurate information for the s...     

Microfluidic integration for electrochemical biosensor applications

Electrochemical biosensors are particularly suitable for miniaturization and integration in microfluidic devices. Applications include the detection of whole cells, cell components, proteins, and small molecules to address tasks in the fields of diagnostics and food and environmental control. Microfluidic setups range from simple channels for sample transport to channels with integrated sensing electrodes to highly sophisticated platforms with additional elements for sample preparation. The design of the microfluidics depends on both the type of detection and on the application and sample material. This review summarizes recent work on electrochemical biosensors with integrated microfluidics with the focus on developments for real sample applications, particularly those including measurements with real sample media.     

微流控芯片技术在生命科学研究中的应用

微流控芯片最初起源于分析化学领域,是一种采用精细加工技术,在数平方厘米的基片,制作出微通道网络结构及其它功能单元,以实现集微量样品制备、进样、反应、分离及检测于一体的快速、高效、低耗的微型分析实验装置.随着微电子及微机械制作技术的不断进步,近年来微流控芯片技术发展迅猛,并开始在化学、生命科学及医学器件等领域发挥重要作用.本文首先简单介绍了微流控芯片制作材料和工艺,然后主要阐述了其在蛋白质分离、免疫分析、DNA分析和测序、细胞培养及检测等方面的应用进展.     

微流控技术在外泌体分离分析中的研究进展

外泌体是一类由细胞分泌的含有脂质、蛋白、核酸等多种物质的纳米级囊泡,主要参与细胞间的物质交换及信息传导,与多种疾病的发生发展密切相关。对外泌体进行深入研究,理解其生物学功能,对疾病诊断与治疗具有重要意义。由于外泌体尺寸较小且密度和体液接近,想要对复杂生物样本中的外泌体进行分离与分析十分困难。传统的外泌体分离方法如超速离心、超滤等大都需要借助大型仪器设备,且耗时长、操作复杂。因此迫切需要开发高效、便捷的外泌体分离检测手段。微流控技术因其微型化、高通量、可集成等特点,为外泌体的分离分析提供了一个新的平台。该文主要对近年来微流控技术在外泌体分离分析相关领域的研究进展进行了综述。重点从外泌体物理特性和生化特性两个角度出发,介绍了微流控芯片技术用于外泌体分离领域的主要原理、策略和方法。此外,还介绍了微流控技术与荧光、电化学传感、表面等离子体共振等多模态检测方法结合,实现外泌体一体化分析的新进展。最后,该文分析了目前微流控技术用于外泌体分离检测存在的挑战,并对其发展趋势和前景进行了展望。随着微流控外泌体分离分析装置的不断微型化、集成化、自动化,微流控芯片技术将在外泌体分离、生化检测、机制研究等方面将发挥越来越重要的作用。     

基于微流控技术的细胞水平高通量药物筛选系统的研究进展

药物筛选是新药研发的关键步骤,创新药物的发现需要采用适当的药物作用靶点对大量化合物样品进行筛选.高通量筛选系统能够实现数千个反应同时测试和分析,大大提高了药物筛选的实验规模和效率.其中基于细胞水平的高通量药物筛选系统因为更加接近人体生理条件,成为主要的筛选模式.而目前发展成熟的高通量细胞筛选系统主要基于多孔板,存在细胞...     

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.     

Integrated electrofluidic circuits: pressure sensing with analog and digital operation functionalities for microfluidics

Microfluidic technology plays an essential role in various lab on a chip devices due to its desired advantages. An automated microfluidic system integrated with actuators and sensors can further achieve better controllability. A number of microfluidic actuation schemes have been well developed. In contrast, most of the existing sensing methods still heavily rely on optical observations and external transducers, which have drawbacks including: costly instrumentation, professional operation, tedious interfacing, and difficulties of scaling up and further signal processing. This paper reports the concept of electrofluidic circuits - electrical circuits which are constructed using ionic liquid (IL)-filled fluidic channels. The developed electrofluidic circuits can be fabricated using a well-developed multi-layer soft lithography (MSL) process with polydimethylsiloxane (PDMS) microfluidic channels. Electrofluidic circuits allow seamless integration of pressure sensors with analog and digital operation functions into microfluidic systems and provide electrical readouts for further signal processing. In the experiments, the analog operation device is constructed based on electrofluidic Wheatstone bridge circuits with electrical outputs of the addition and subtraction results of the applied pressures. The digital operation (AND, OR, and XOR) devices are constructed using the electrofluidic pressure controlled switches, and output electrical signals of digital operations of the applied pressures. The experimental results demonstrate the designed functions for analog and digital operations of applied pressures are successfully achieved using the developed electrofluidic circuits, making them promising to develop integrated microfluidic systems with capabilities of precise pressure monitoring and further feedback control for advanced lab on a chip applications.     

Microfluidic chips for mass spectrometry‐based proteomics

Microfluidic devices coupled to mass spectrometers have emerged as excellent tools for solving the complex analytical challenges associated with the field of proteomics. Current proteome identification procedures are accomplished through a series of steps that require many hours of labor‐intensive work. Microfluidics can play an important role in proteomic sample preparation steps prior to mass spectral identification such as sample cleanup, digestion, and separations due to its ability to handle small sample quantities with the potential for high‐throughput parallel analysis. To utilize microfluidic devices for proteomic analysis, an efficient interface between the microchip and the mass spectrometer is required. This tutorial provides an overview of the technologies and applications of microfluidic chips coupled to mass spectrometry for proteome analysis. Various approaches for combining microfluidic devices with electrospray ionization (ESI) and matrix‐assisted laser desorption/ionization (MALDI) are summarized and applications of chip‐based separations and digestion technologies to proteomic analysis are presented. Copyright © 2009 John Wiley & Sons, Ltd.     

Continuous flow multi-stage microfluidic reactors via hydrodynamic microparticle railing

"Multi-stage" fluidic reactions are integral to diverse biochemical assays; however, such processes typically require laborious and time-intensive fluidic mixing procedures in which distinct reagents and/or washes must be loaded sequentially and separately (i.e., one-at-a-time). Microfluidic processors that enable multi-stage fluidic reactions with suspended microparticles (e.g., microbeads and cells) to be performed autonomously could greatly extend the efficacy of lab-on-a-chip technologies. Here we present a single-layer microfluidic reactor that utilizes a microfluidic railing methodology to passively transport suspended microbeads and cells into distinct, adjacent laminar flow streams for rapid fluidic mixing and assaying. Four distinct molecular synthesis processes (i.e., consisting of 48 discrete fluidic mixing stages in total) were accomplished on polystyrene microbead substrates (15 μm in diameter) in parallel, without the need for external observation or regulation during device operation. Experimental results also revealed successful railing of suspended bovine aortic endothelial cells (approximately 13 to 17 μm in diameter). The presented railing system provides an effective continuous flow methodology to achieve bead-based and cell-based microfluidic reactors for applications including point-of-care (POC) molecular diagnostics, pharmacological screening, and quantitative cell biology.     

Dose-dependent cell-based assays in V-shaped microfluidic channels

The capability of lab-on-a-chip technologies in controlling cell transportation, generating concentration gradients, and monitoring cellular responses offers an opportunity to integrate dose-dependent cell-based bioassays on a chip. In this study, we have developed microfluidic modules featured with channel components and sandbag structures for positioning biological cells within the microchip. We have demonstrated that by geometric modulation of the microchannel architectures, it is possible to immobilize individual cells at desired locations with controllable numbers, to generate defined concentration gradients at various channel lengths, and to improve the efficiency and reproducibility in data acquisition. The microfluidic module was used to exercise a series of cell-based assays, including the measurement of kinetics and dynamics of intracellular enzymatic activities, the analysis of cellular response under the stimulation of two chemicals with defined concentration profiles, and the study of laser irradiation effect on cellular uptake of photosensitizers. The results demonstrated the capabilities of the microfluidic module for simultaneously conducting multiple sets of dose-dependent, cell-based bioassays, and for quantitatively comparing responses of individual cells under various stimulations.     

Array-based capture, distribution, counting and multiplexed assaying of beads on a centrifugal microfluidic platform

We present a novel centrifugal microfluidic platform for the highly efficient manipulation and analysis of particles for applications in bead-based assays. The platform uses an array of geometrical V-cup barriers to trap particles using stopped-flow sedimentation under highly reproducible hydrodynamic conditions. The impact parameters governing the occupancy distribution and capture efficiency of the arrayed traps are investigated. The unique, nearly 100% capture efficiency paired with the capability to establish sharply peaked, single occupancy distributions enables a novel, digital readout mode for color-multiplexed, particle-based assays with low-complexity instrumentation. The presented technology marks an essential step towards a versatile platform for the integration of bead- and cell-based biological assays.     

微流控细胞芯片生命分析应用多元化

微流控芯片是现代生命科学研究领域的重要分析工具.结合研究者近年来开展的研究工作和取得的相关进展,本文主要介绍了微流控细胞芯片的功能特征,同时从动物细胞、植物细胞以及微生物细胞三方面系统阐述了微流控芯片生命分析多元化的发展现状,并对其应用前景进行了展望.