Digital microfluidics for cell-based assays

We introduce a new method for implementing cell-based assays. The method is based on digital microfluidics (DMF) which is used to actuate nanolitre droplets of reagents and cells on a planar array of electrodes. We demonstrate that this method is advantageous for cell-based assays because of automated manipulation of multiple reagents in addition to reduced reagent use and analysis time. No adverse effects of actuation by DMF were observed in assays for cell viability, proliferation, and biochemistry. A cytotoxicity assay using Jurkat T-cells was performed using the new method, which had approximately 20 times higher sensitivity than a conventional well plate assay. These results suggest that DMF has great potential as a simple yet versatile analytical tool for implementing cell-based assays on the microscale.     

微流控液滴技术:微液滴生成与操控

微液滴技术因具有高通量两相分割分离能力,吸引众多不同领域研究者的关注.本文回顾了微流控液滴技术领域的一些基本技术思路,涉及微液滴的流控生成方法,包括水动力法、电动法、气动法、光控法等,以及液滴生成后的操控技术,如液滴定向位移、融合、裂分、混合、分选、捕获等,同时对这些方法作了简要评述.     

A feedback control system for high-fidelity digital microfluidics

Digital microfluidics (DMF) is a technique in which discrete droplets are manipulated by applying electrical fields to an array of electrodes. In an ideal DMF system, each application of driving potential would cause a targeted droplet to move onto an energized electrode (i.e., perfect fidelity between driving voltage and actuation); however, in real systems, droplets are sometimes observed to resist movement onto particular electrodes. Here, we implement a sensing and feedback control system in which all droplet movements are monitored, such that when a movement failure is observed, additional driving voltages can be applied until the droplet completes the desired operation. The new system was evaluated for a series of liquids including water, methanol, and cell culture medium containing fetal bovine serum, and feedback control was observed to result in dramatic improvements in droplet actuation fidelity and velocity. The utility of the new system was validated by implementing an enzyme kinetics assay with continuous mixing. The new platform for digital microfluidics is simple and inexpensive and thus should be useful for scientists and engineers who are developing automated analysis platforms.     

Microfluidics technology for manipulation and analysis of biological cells

Analysis of the profiles and dynamics of molecular components and sub-cellular structures in living cells using microfluidic devices has become a major branch of bioanalytical chemistry during the past decades. Microfluidic systems have shown unique advantages in performing analytical functions such as controlled transportation, immobilization, and manipulation of biological molecules and cells, as well as separation, mixing, and dilution of chemical reagents, which enables the analysis of intracellular parameters and detection of cell metabolites, even on a single-cell level. This article provides an in-depth review on the applications of microfluidic devices for cell-based assays in recent years (2002–2005). Various cell manipulation methods for microfluidic applications, based on magnetic, optical, mechanical, and electrical principles, are described with selected examples of microfluidic devices for cell-based analysis. Microfluidic devices for cell treatment, including cell lysis, cell culture, and cell electroporation, are surveyed and their unique features are introduced. Special attention is devoted to a number of microfluidic devices for cell-based assays, including micro cytometer, microfluidic chemical cytometry, biochemical sensing chip, and whole cell sensing chip.     

A review of chemical gradient systems for cell analysis

Microfluidic spatial and temporal gradient generators have played an important role in many biological assays such as in the analysis of wound healing, inflammation, and cancer metastasis. Chemical gradient systems can also be applied to other fields such as drug design, chemical synthesis, chemotaxis, etc. Microfluidic systems are particularly amenable to gradient formation, as the length scales used in chips enable fluid processes that cannot be conducted in bulk scale. In this review we discuss new microfluidic devices for gradient generation and applications of those systems in cell analysis.     

Mimic Nature Using Chemotaxis of Ionic Liquid Microdroplets for Drug Delivery Purposes

Due to the growing prevalence of incurable diseases, such as cancer, worldwide, nowadays, the development of smart drug delivery systems is an inevitable necessity. Chemotaxis-driven movement of ionic liquid microdroplets containing therapeutic compounds is a well-known example of a smart drug delivery system. This review aims to classify, summarize, and compare ionic liquid-based chemotaxis systems in an easily understandable article. Chemotaxis is the basis of the movement of cells and microorganisms in biological environments, which is the cause of many vital biochemical and biological processes. This review attempts to summarize the available literature on single-component biomimetic and self-propelling microdroplet systems based on ionic liquids, which exhibit chemotaxis and spontaneously move in a determined direction by an external gradient, particularly a chemical change. It also aims to review artificial ionic liquid-based chemotaxis systems that can be used as drug carriers for medical purposes. The various ionic liquids used for this purpose are discussed, and different forms of chemical gradients and mechanisms that cause movement in microfluidic channels will be reviewed.     

Optically-actuated translational and rotational motion at the microscale for microfluidic manipulation and characterization

The single beam optical trap (optical tweezers), a highly focused beam, is on its way to revolutionizing not only the fields of colloidal physics and biology, but also materials science and engineering. Recently, spatially-extended three-dimensional light patterns have gained considerable usage for exerting force to alter, manipulate, organize and characterize materials. To advance the degree of manipulation, such as rotation of materials in microfluidic environments along with spatial structuring, other beam parameters such as phase and polarization have to be configured. These advances in optical tweezers' technology have enabled complex microfluidic actuation and sorting. In addition to remotely (in a non-contact way) applying force and torques in three-dimensions, which can be continuously varied unlike mechanical manipulators, optical tweezers-based methods can be used for sensing the force of interaction between microscopic objects in a microfluidic environment and for the characterization of micro-rheological properties. In this review, we place emphasis on applications of optical actuation based on novel beams in performing special functions such as rotation, transportation, sorting and characterization of the microscopic objects. Further, we have an extended discussion on optical actuation (transport and rotation) with fiber optic microbeams and spectroscopic characterization in the microfluidic environment. All these advancements in optical manipulation would further facilitate the growing use of optical tools for complex microfluidic manipulations.     

Pluronic additives: a solution to sticky problems in digital microfluidics

Digital microfluidics (DMF) is a promising technique for carrying out miniaturized, automated biochemical assays in which discrete droplets of reagents are actuated on the surface of an array of electrodes. A limitation for DMF is nonspecific protein adsorption to device surfaces, which interferes with assay fidelity and can cause droplets to become unmovable. Here, we report the results of a quantitative analysis of protein adsorption on DMF devices by means of confocal microscopy and secondary ion mass spectrometry. This study led us to a simple and effective method for limiting the extent of protein adsorption: the use of low concentrations of Pluronic F127 as a solution additive. This strategy has a transformative effect on digital microfluidics, facilitating the actuation of droplets containing greater than 1000-fold higher protein concentrations than is possible without the additive. To illustrate the benefits of this new method, we implemented a DMF-driven protein digest assay using large concentrations (1 mg/mL) of protein-substrate. The use of Pluronic additives solves a sticky problem in DMF, which greatly expands the range of applications that are compatible with this promising technology.     

响应性交联液晶高分子仿生致动器的研究进展

交联液晶高分子兼具液晶取向有序性和交联聚合物熵弹性等特点,能够以动态可调节和可逆的方式来模仿生物体的行为,在仿生器件、柔性机器人、智能表面、生物医药等领域具有良好的应用前景.本综述总结了近几年智能响应性交联液晶高分子在仿生致动器方面的研究进展,从响应性交联液晶高分子的结构和驱动机理出发,讨论了响应性交联液晶高分子的合成工艺、制备技术和成型方法,以及响应性交联液晶高分子对光、热、磁、湿度的响应.此外,介绍了响应性交联液晶高分子致动器在柔性机器人、人工肌肉、微流体运输等领域的应用.最后,对响应性交联液晶高分子的发展前景进行了展望.这项工作主要讨论了响应性交联液晶高分子,旨在为具有新颖功能和更有挑战性的智能微型致动器提供新的设计思路.     

基于SAFT理论关联醇与DMF体系的1H NMR

混合物核磁共振化学位移可以用不同的化学缔合理论研究,但当形成的缔合体种类很多时,需要很多优化参数.作者从统计缔合流体理论（SAFT）出发,提出了一个能关联混合物核磁共振化学位移,但又不需要假设缔合平衡常数的模型.对于醇与N,N-二甲基甲酰胺（DMF）体系,关联的均方根偏差小于1.01％.讨论了醇与DMF体系和醇与正己烷体系核磁共振化学位移随醇的浓度变化趋势的差异,认为醇与DMF形成比醇的自缔更强的交叉缔合是造成这种变化趋势不同的主要原因.     

Magnetization of individual yeast cells by in situ formation of iron oxide on cell surfaces

Magnetic functionalization of living cells has intensively been investigated with the aim of various bioapplications such as selective separation, targeting, and localization of the cells by using an external magnetic field. However, the magnetism has not been introduced to individual living cells through the in situ chemical reactions because of harsh conditions required for synthesis of magnetic materials. In this work, magnetic iron oxide was formed on the surface of living cells by optimizing reactions conditions to be mild sufficiently enough to sustain cell viability. Specifically, the reactive LbL strategy led to formation of magnetically responsive yeast cells with iron oxide shells. This facile and direct post-magnetization method would be a useful tool for remote manipulation of living cells with magnetic interactions, which is an important technique for the integration of cell-based circuits and the isolation of cell in microfluidic devices.     

One-dimensional magnetic inorganic-organic hybrid nanomaterials

One-dimensional (1D) magnetic inorganic-organic hybrid nanomaterials bear both the intrinsic magnetic properties of the inorganic components and the functionality and responsiveness of their organic part. In this tutorial review, we first emphasize various synthetic strategies for this type of materials: (i) template-directed synthesis employs different preformed templates such as channels in solids, mesostructures self-assembled from block copolymers, cylindrical polymer brushes, 1D biological templates and other existing 1D templates; (ii) electrospinning, which provides a simple and efficient technique that can lead to a potential large-scale production; (iii) 1D conjugation of building blocks which combines the physical attraction of magnetic nanoparticles in a magnetic field with chemical crosslinking and stabilization. The properties, functions and the future trends of these materials are also briefly introduced. It is foreseeable that these hybrid materials will play more and more important roles in the ever-advancing miniaturization of functional devices.     

Rational pathway engineering of type I fatty acid synthase allows the biosynthesis of triacetic acid lactone from D-glucose in vivo

Metabolic pathway engineering is a powerful tool to synthesize structurally diverse and complex chemicals via genetic manipulation of multistep catalytic systems involved in cell metabolism. Here, we report the rational design of a fatty acid biosynthetic pathway, Brevibacterium ammoniagenes fatty acid synthase B (FAS-B), that allows the microbial synthesis of triacetic acid lactone (TAL) from an inexpensive feedstock, d-glucose. TAL can be chemically converted to phloroglucinol, which is a core structure for the synthesis of various high value bioactive compounds and energetic compounds such as 1,3,5-triamino-2,4,6-trinitrobenzene (TATB). Synthesis of phloroglucinol from d-glucose using this combined biological and chemical synthesis may offer significant advantages over the current phloroglucinol manufacture, including environmental friendliness and reduction in the cost of phloroglucinol. More importantly, it represents a novel strategy for the benzene-free synthesis of aromatic chemicals.     

New sorbents for extraction and microextraction techniques

This review outlines recent progress in the research on some new classes of sorbents for extraction and microextraction techniques. Carbon nanotubes are allotropes of carbon with cylindrical structure. They are very stable systems having considerable chemical inertness due to the strong covalent bonds of the carbon atoms on the nanotube surface. Some applications of carbon nanotubes are presented in a perspective view. Molecular imprinting has proved to be an effective technique for the creation of recognition sites on a polymer scaffold. By a mechanism of molecular recognition, the molecularly imprinted polymers are used as selective tools for the development of various analytical techniques such as liquid chromatography, capillary electrochromatography, solid-phase extraction (SPE), binding assays and biosensors. Sol–gel chemistry provides a convenient pathway to create advanced material systems that can be effectively utilized to solve the solid phase microextraction fiber technology problems. This review is mainly focused on recent advanced developments in the design, synthesis and application of sol–gel in preparation of coatings for the SPME fibers.     

连续流动式聚合酶链式反应生物芯片/微装置中脱氧核糖核酸样品的驱动控制技术进展

介绍国内外连续流动式聚合酶链式反应生物芯片／微装置中脱氧核糖核酸样品的驱碧控制技术进展，主要包括恒流泵（注射泵驱动和蠕动泵驱动）、旋转泵驱动、磁流体动力驱动以及自然对流驱动等。并对这几种驱动方式的优缺点作简要的讨论（引用文献43篇）。     

A digital microfluidic method for multiplexed cell-based apoptosis assays

Digital microfluidics (DMF), a fluid-handling technique in which picolitre-microlitre droplets are manipulated electrostatically on an array of electrodes, has recently become popular for applications in chemistry and biology. DMF devices are reconfigurable, have no moving parts, and are compatible with conventional high-throughput screening infrastructure (e.g., multiwell plate readers). For these and other reasons, digital microfluidics has been touted as being a potentially useful new tool for applications in multiplexed screening. Here, we introduce the first digital microfluidic platform used to implement parallel-scale cell-based assays. A fluorogenic apoptosis assay for caspase-3 activity was chosen as a model system because of the popularity of apoptosis as a target for anti-cancer drug discovery research. Dose-response profiles of caspase-3 activity as a function of staurosporine concentration were generated using both the digital microfluidic method and conventional techniques (i.e., pipetting, aspiration, and 96-well plates.) As expected, the digital microfluidic method had a 33-fold reduction in reagent consumption relative to the conventional technique. Although both types of methods used the same detector (a benchtop multiwell plate reader), the data generated by the digital microfluidic method had lower detection limits and greater dynamic range because apoptotic cells were much less likely to de-laminate when exposed to droplet manipulation by DMF relative to pipetting/aspiration in multiwell plates. We propose that the techniques described here represent an important milestone in the development of digital microfluidics as a useful tool for parallel cell-based screening and other applications.     

Molecularly defined graphitic interface toward proton manipulation

Proton plays a critical role in electrochemical systems to control electrochemical reactivity or isotopic enrichment. Graphene is intensively investigated owing to its unique electronic structure and device fabrication. Through the structural tunability of graphitic materials by chemical or physical modification of the surface, graphene is revealed to be an ideal material for proton manipulation. Here, we review the use of graphene or graphitic materials toward the manipulation of proton with regard to the following three points. (1) Electronic properties of graphene: The electronic band structure of graphene can be modified by metal contacts owing to the interaction with a metal surface. (2) Molecular control of graphitic interface: The chemical structure of graphene can be modified, as is done in molecular chemistry, and can be used as a catalytic platform. (3) Proton conduction by graphene: Proton transport through a graphene layer occurs with a unique mechanism such as tunneling. We provide a perspective on the use of graphitic materials toward controlling the behavior of protons on the basis of the aforementioned points. From the above, graphene can be used as a platform for proton manipulation.     

Liquid interfacial nanoarchitectonics: Molecular machines,organic semiconductors,nanocarbons, stem cells,and others

The concept of nanoarchitectonics has been proposed as an extensional development of nanotechnology through fusions with material science and the other fields. In nanoarchitectonics, nano-units of atoms, molecules, and nanomaterials are architected into construction of functional material systems. In order to assemble intended structures or hierarchical structures from nano-units, it is more useful to confine nano-units at the interface. In addition, nanoarchitectonics is expected to output functions by harmonizing many units in dynamic environments. However, the liquid interfaces still have lots of unexplored matters in nanoscale because supports by advanced apparatus and techniques in nanotechnology are not always available. Specifically, this review paper summarizes examples of research on molecular manipulation, molecular arrangement and assembly, materials synthesis, and life manipulation at the liquid interface. These examples demonstrate that the liquid interface enables the control of dynamic functions of various size regions, from molecular-level phenomena such as the control of molecular machines to techniques of living creature size such as the control of stem cell differentiation. Liquid interfaces are very useful environments for controlling dynamic functions for a wide range of targets and would have tremendous potential in terms of functional exploration. The great potential of nanoarchitectonics at the liquid interface and the challenges to be solved in the future are also discussed.     

Chemical Synthesis at Surfaces with Atomic Precision: Taming Complexity and Perfection

Scanning probe microscopy (SPM) is a powerful tool to study the structure and dynamics of molecules at surfaces and interfaces as well as to precisely manipulate atoms and molecules by applying an external force, by inelastic electron tunneling, or by means of an electric field. The rapid development of these SPM manipulation modes made it possible to achieve fine‐control over fundamental processes in the physics of interfaces as well as chemical reactivity, such as adsorption, diffusion, bond formation, and bond dissociation with precision at the single atom/molecule level. Their controlled use for the fabrication of atomic‐scale structures and synthesis of new, perhaps uncommon, molecules with programmed properties are reviewed. Opportunities and challenges towards the development of complex chemical systems are discussed, by analyzing potential future impacts in nanoscience and nanotechnology.     

Beyond the spin model: exchange coupling in molecular magnets with unquenched orbital angular momenta

In this critical review we review the problem of exchange interactions in polynuclear metal complexes involving orbitally degenerate metal ions. The key feature of these systems is that, in general, they carry an unquenched orbital angular momentum that manifests itself in all their magnetic properties. Thus, interest in degenerate systems involves fundamental problems related to basic models in magnetism. In particular, the conventional Heisenberg-Dirac-Van Vleck model becomes inapplicable even as an approximation. In the first part we attempt to answer two key questions, namely which theoretical tools are to be used in the case of degeneracy, and how these tools can be employed. We demonstrate that the exchange interaction between orbitally degenerate metal ions can be described by the so-called orbitally-dependent exchange Hamiltonian. This approach has shown to reveal an anomalously strong magnetic anisotropy that can be considered as the main physical manifestation of the unquenched orbital angular momentum in magnetic systems. Along with the exchange coupling, a set of other interactions (such as crystal field effects, spin-orbit and Zeeman coupling), which are specific for the degenerate systems, need to be considered. All these features will be discussed in detail using a pseudo-spin-1/2 Hamiltonian approach. In the second part, the described theoretical background will be used to account for the magnetic properties of several magnetic metal clusters and low-dimensional systems: (i) the dinuclear face-sharing unit [Ti(2)Cl(9)](3-), which exhibits a large magnetic anisotropy; (ii) the rare-earth compounds Cs(3)Yb(2)Cl(9) and Cs(3)Yb(2)Br(9), which, surprisingly, exhibit a full magnetic isotropy; (iii) a zig-zag Co(II) chain exhibiting unusual combination of single-chain magnet behavior and antiferromagnetic exchange coupling; (iv) a trigonal bipyramidal Ni(3)Os(2) complex; (v) various Co(II) clusters encapsulated by polyoxometalate ligands. In the two last examples a pseudospin-1/2 Hamiltonian approach is applied to account for the presence of exchange anisotropy (150 references).