Probing single cells using flow in microfluidic devices

Enabling fluids to be manipulated on the micron-scale, microfluidic technologies have facilitated major advances in how we study cells. In this review, we highlight key developments in how flow in microfluidic devices is exploited to investigate the behavior of individual cells, from trapping and positioning single cells to probing cell deformability. Exploiting the properties of fluids and flow patterns in microchannels makes it possible to study large populations of single cells at micron-length scales with increased throughput and efficiency.     

Chladni figures revisited based on nanomechanics

Chladni patterns based on nanomechanics in the microfluidic environment are presented. In contrast with the macroscopic observations in the gaseous environment, nanoparticles are found to move to the nodes, whereas micron-sized particles move to the antinodes of the vibrating interface. This opens the door to size-based sorting of particles in microfluidic systems, and to highly parallel and controlled assembly of biosensors and nanoelectronic circuits.     

Fluorescence lifetime imaging of mixing dynamics in continuous-flow microdroplet reactors

Water-in-oil microdroplets within fluidic channels have the potential to serve as isolated reaction compartments for monitoring real-time dynamics with high efficiency and repeatability. Droplets, usually generated from aqueous and oil solutions using standard microfluidic formats, can be produced at frequencies in excess of 1 kHz. Although mixing within such microdroplets is normally enhanced by chaotic advection, the mixing pattern from droplet to droplet is almost identical and reproducible in form. Herein, we demonstrate that fluorescence lifetime imaging can be used to reconstruct mixing patterns within a droplet with a time resolution of 5 micros.     

Microwave bonding of poly(methylmethacrylate) microfluidic devices using a conductive polymer

Component binding within microfluidic devices is a problem that has long been seeking a solution. In this investigation, the use of microwave radiation to seal PMMA components has been investigated using polyaniline as an absorber that is capable of inducting interfacial bonding. Straight microchannels were machined into PMMA using a Datron CAT3DM6 CNC machine with widths and depths across a range of 100-1000 μm. Prototype fluidic devices were prepared with channel patterns utilizing varying feature sizes, bends and flow profiling to demonstrate the application of the technique to real microfluidic devices. Experimental data illustrated the successful bonding of channels in the range stated previously and bonding (tensile) strength was assessed via pull tests on bonded PMMA using an Engstrom Zwick 100 tensile testing system (Engstrom Ltd, US). Coherent, defect free seals were attained with breakage tests requiring an excess of 1 kN force.     

Microfluidic colloidal island formation and erasure induced by surface acoustic wave radiation

Spatiotemporal patterns form in many nonlinear physicochemical or biological systems. Although unusual, microfluidic systems are no exception. We observe such patterns to form by colloids along the free surface of a drop beneath which surface acoustic waves propagate, and propose fundamental mechanisms to elucidate their formation. With increasing excitation amplitude, the colloids first assemble into concentric rings and then cluster into islands due to a combination of capillarity and surface acceleration. As the excitation is further increased, fluid streaming commences within the drop, inducing a transient metastable state in which the system alternates between colloidal island formation on the quiescent drop surface and subsequent erasure due to local vortex generation.     

4-D imaging of fluid flow with digital in-line holographic microscopy

The large depth of field of digital in-line holographic microscopy (DIHM) with numerical reconstruction provides an ideal tool for the study of microfluidic phenomena. As indicators of the flow patterns we use latex microspheres and also red blood cells whose three-dimensional trajectories and velocities can easily be measured as a function of time with subsecond and micron resolution. We demonstrate the efficiency of DIHM by showing 3-D views of the flow patterns around big spheres in various geometric arrangements.     

Investigating passive immunodiffusion reactions in the channels of a microfluidic device

The passive immunodiffusion reaction in a microfluidic system is investigated via the interaction between fluorescein human albumin conjugate and monoclonal antibodies. The topology of a microfluidic device suitable for this reaction was designed, and the device was fabricated using LIGA technology. A system for transitioning from the macro- to the microlevel is developed and produced. The dissociation constants and specificity of interaction in complexes formed as a result of the reaction between fluorescein human albumin conjugates and monoclonal antibodies are determined. The results from our study reveal that systems based on combinations of passive immunodiffusion reactions and microfluidic technologies are applicable for detecting antigens and antibodies in the analyzed fluids.     

Water-assisted femtosecond laser ablation for fabricating three-dimensional microfluidic chips

When the femtosecond laser focused in the water, the breakdown will be induced. The generated high-speed jet and shock wave can be used to etch silica glass for fabricating three-dimensional (3D) microfluidic chips. We present a simple and practical method to produce 3D multilayer microfluidic chips in silica glass. This method offers high design flexibility and fabricating feasibility. We also introduce a convenient cleaning method for diluting and ejecting the ablated debris from microchannel. Therefore, the femtosecond laser induced high-speed jet and shock wave can be used to fabricate complex microfluidic chips in silica glass. Experimental results show that the diameter of microchannel is uniform and the complexity of the microfluidic chip is under control. As a proof of principle, we demonstrate the feasibility of the fabricating process by using the water-assisted femtosecond laser ablation.     

Microfluidic mixing using periodically induced secondary potential in electroosmotic flow

This research develops a novel microfluidic mixing in a simple manner: An electrode was installed between two primary electrodes that generate DC fields and induce electroosmotic flow (EOF). While EOF was introduced by DC fields from the two primary electrodes, the active mixing could be achieved if potential was supplied to the secondary anode. Normal EOF was recovered as soon as the supply of potential to the secondary anode was terminated. By periodically repeating the on/off phase of the driving potential to the secondary anode, oscillating and vortex swirling flow patterns occurred and vigorous and efficient mixing was obtained.     

飞秒激光直写PMMA制备微流道的工艺技术研究

针对目前PMMA微流道加工质量差和效率低的问题, 对飞秒激光直写PMMA制备微流道的工艺技术进行了研究。通过实验分析了不同激光参数对微流道的宽度、深度、粗糙度、微流道两侧堆积物火山口高度的影响及变化规律。实验结果表明, 当激光扫描速度为20 mm/s时, 激光功率为0.5 W时, 微流道粗糙度较低且变化幅度不明显; 激光能量从0.5 W增加到0.75 W时, 微流道的宽度、深度与激光能量呈线性关系增加; 激光功率大于0.5 W时, 随着激光功率以及加工次数的增加, 微流道宽度、深度、粗糙度以及堆积物火山口的高度逐渐增加。经过计算得出, PMMA的烧蚀阈值为0.357 J/cm2。通过优化工艺参数, 制备出粗糙度较低、表面光滑、深度为16 μm的微流道芯片。     

Microfluidic laser embedded in glass by three-dimensional femtosecond laser microprocessing

Microfluidic dye lasers three-dimensionally embedded in glass have been fabricated for what is believed to be the first time by integrating micro-optical and microfluidic components by use of a femtosecond laser. By pumping the microfluidic laser, in which the microfluidic chamber was filled with the laser dye Rhodamine 6G dissolved in ethanol, with a frequency-doubled Nd:yttrium aluminum garnet laser, lasing action was confirmed by analysis of the emission spectra at different pump powers. In addition, by arranging two microfluidic chambers serially in the glass, we built a microfluidic twin laser that produces an array of two simultaneous laser emissions with one pump laser.     

Engineering a material for biomedical applications with electric field assisted processing

In this work, using multiple co-flows we demonstrate in-situ encapsulation of nano-particles, liquids and/or gases in different structural morphologies, which can also be deposited in a designated pattern by a direct write method and surface modification can be controlled to release encapsulated material. The range of possibilities offered by exposing a material solution to an applied electric field can result in a plethora of structures which can accommodate a whole host of biomedical applications from microfluidic devices (microchannels, loaded with various materials), printed 3D structures and patterns, lab-on-a-chip devices to encapsulated materials (capsules, tubes, fibres, dense multi-layered fibrous networks) for drug delivery and tissue engineering. The structures obtained in this way can vary in size from micrometer to the nanometer range and the processing is viable for all states of matter. The work shown demonstrates some novel structures and methodologies for processing a biomaterial.     

High-Q microfluidic cavities in silicon-based two-dimensional photonic crystal structures

We demonstrate postprocessed microfluidic double-heterostructure cavities in silicon-based photonic crystal slab waveguides. The cavity structure is realized by selective fluid infiltration of air holes using a glass microtip, resulting in a local change of the average refractive index of the photonic crystal. The microcavities are probed by evanescent coupling from a silica nanowire. An intrinsic quality factor of 57,000 has been derived from our measurements, representing what we believe to be the largest value observed in microfluidic photonic crystal cavities to date.     

Thermophoresis of DNA determined by microfluidic fluorescence

We describe a microfluidic all-optical technique to measure the thermophoresis of molecules. Within micrometer-thick chambers, we heat aqueous solutions with a micrometer-sized focus of infrared light. The temperature increase of about 1 K is monitored with temperature-sensitive fluorescent dyes. We test the approach in measuring the thermophoresis of DNA. We image the concentration of DNA in a second fluorescence-color channel. DNA is depleted away from the heated spot. The profile of depletion is fitted by the thermophoretic theory to reveal the Soret coefficient. We evaluate the method with numerical 3D calculations of temperature profiles, drift, convection and thermophoretic depletion using finite element methods. The approach opens new ways to monitor thermophoresis at the single molecule level, near boundaries and in complex mixtures. The flexible microfluidic setting is a good step towards microfluidic applications of thermophoresis in biotechnology.     

Preparation of nanoparticles by continuous-flow microfluidics

We review a variety of micro- and nanoparticle formulations produced with microfluidic methods. A diverse variety of approaches to generate microscale and nanoscale particles has been reported. Here we emphasize the use of microfluidics, specifically microfluidic systems that operate in a continuous flow mode, thereby allowing continuous generation of desired particle formulations. The generation of semiconductor quantum dots, metal colloids, emulsions, and liposomes is considered. To emphasize the potential benefits of the continuous-flow microfluidic methodology for nanoparticle generation, preliminary data on the size distribution of liposomes formed using the microfluidic approach is compared to the traditional bulk alcohol injection method.     

微流控SERS芯片及其生物传感应用

由于微流控芯片具有优异的集成性和灵活的可操作性,基于芯片上的检测方法被大量开发,发展十分迅速。其中,表面增强拉曼光谱（SERS）凭借其超高的灵敏度、独一无二的指纹谱和窄峰宽等特点成为一种广泛采用的检测手段。SERS微流控芯片集SERS检测技术与微流控芯片的优势于一体,一方面为SERS检测方法的重复性和可靠性提供了一个高效平台,另一方面推动了微流控芯片的功能拓展,在生物分子探测、细胞捕获乃至组织模拟等领域具有广阔的应用前景。本文在简要介绍SERS的原理及其生物传感应用的基础上,重点概述了SERS微流控芯片的构建及其在生物传感及检测中的应用,最后探讨了该研究方向存在的问题及发展方向。     

基于微流控技术的水的太赫兹吸收特性研究

很多生物大分子的特征振动模式和转动模式都位于太赫兹波段范围内,且太赫兹波的低电子能特性使其在实验过程中不会对待测样品造成破坏,所以可以采用太赫兹技术来鉴别生物样品。在许多研究中,生物样品都是溶液状态,溶液中水和其他分子之间的相互作用涉及很多生物现象,所以研究水的太赫兹特性就显得至关重要。众所周知,水分子是十分常见的极性分子,分子间氢键会与太赫兹波发生强烈的相互作用,从而使得水对太赫兹波有很强的吸收作用,导致利用太赫兹技术研究水溶液中生物样品的动态特性变得相当困难。为了解决这一难题,可以引入微流控技术。微流控技术以能精确操控微尺度流体而著称,其沟道深度可以达到50μm甚至更小。由于微流控技术减小了太赫兹波在流体中的传播距离,从而极大地减小了水对太赫兹波的吸收。本研究采用对太赫兹波具有高透过率的Zeonor 1420R材料制成了夹心式微流控芯片,芯片上微沟道的长度、宽度和深度分别为3 cm,4 mm和50 μm,太赫兹探测区的直径为3 mm。在制作微流控芯片时,利用厚度为50μm的强黏性双面胶代替传统夹心式微流控芯片中的聚二甲基硅氧烷(PDMS)薄膜,使微流控芯片在加热过程中不再有漏液现象。另外,设计了一个温控系统,它由加热片、温度传感器和温控仪构成,该温控系统能够以0.1 ℃的精度控制温度。利用该系统对微流控芯片中的去离子水进行加热,从20～90 ℃每隔5 ℃进行一次太赫兹透射测量,通过对实验数据的分析,发现随着温度升高,水的太赫兹透过率不断减小,说明水对太赫兹波的吸收随着温度的升高而变大。此结果为未来在不同环境温度下利用微流控技术研究液态样品的太赫兹吸收特性提供了先决条件,为未来太赫兹的应用与发展提供技术支持。     

微流控SERS及其在生物医学应用的研究进展

微流控芯片以其对微量样品的精确操控能力而引起特别关注,表面增强拉曼光谱(SERS)作为一种超灵敏的光谱检测技术近年来在痕量检测上应用广泛。微流控芯片与SERS相结合的系统可对微量生物样品进行无损、快速、高灵敏度且高通量的检测分析,在生物医学领域有巨大的应用潜力,是当前的研究热点之一。本文介绍了微流控SERS系统近年的发展情况,包括微流控芯片的制作加工和流体操控,以及微流控芯片中SERS基底的集成;并重点介绍了近年来SERS微流控芯片系统在生物医学上的应用,如生物分子的检测、细胞分析、药物监测和筛选、疾病诊断,以及在环境和食品健康安全方面的检测应用。     

Time-of-flight flow imaging of two-component flow inside a microfluidic chip

Here we report on using NMR imaging and spectroscopy in conjunction with time-of-flight tracking to noninvasively tag and monitor nuclear spins as they flow through the channels of a microfluidic chip. Any species with resolvable chemical-shift signatures can be separately monitored in a single experiment, irrespective of the optical properties of the fluids, thereby eliminating the need for foreign tracers. This is demonstrated on a chip with a mixing geometry in which two fluids converge from separate channels, and is generally applicable to any microfluidic device through which fluid flows within the nuclear spin-lattice relaxation time.     

电场对协流式微流控装置中乳液液滴生成行为的调控机理

建立了直流电场作用下协流式微流控装置中单乳液液滴乳化生成过程的非稳态理论模型,并开展了数值模拟研究,揭示了电场对液滴乳化生成动力学行为的调控机理,阐明了流场/电场参数对液滴乳化生成特性的影响规律.研究结果表明:沿流体流动方向施加静电场可在电物性参数不同的两相流体界面法线方向上产生指向内相流体的电场力,进而强化了内相流体界面的颈缩和断裂,提升了液滴生成速率和形变程度,减小了液滴生成尺寸;在同一毛细数下,随着电毛细数的增大,乳液乳化流型由每周期仅有单一液滴生成的滴式流型转变为每周期有一个主液滴并伴随有卫星液滴生成的滴式流型;随着毛细数和电毛细数的增大,黏性拖曳力以及电场力作用增强,使内相流体颈缩过程后期更容易形成细长型液线,从而有助于诱发液线上产生Rayleigh-Plateau不稳定现象,继而促进卫星液滴的形成.