Particle separation and sorting in microfluidic devices: a review

Separation and sorting of micron-sized particles has great importance in diagnostics, chemical and biological analyses, food and chemical processing and environmental assessment. By employing the unique characteristics of microscale flow phenomena, various techniques have been established for fast and accurate separation and sorting of microparticles in a continuous manner. The advancements in microfluidics enable sorting technologies that combine the benefits of continuous operation with small-sized scale suitable for manipulation and probing of individual particles or cells. Microfluidic sorting platforms require smaller sample volume, which has several benefits in terms of reduced cost of reagents, analysis time and less invasiveness to patients for sample extraction. Additionally, smaller size of device together with lower fabrication cost allows massive parallelization, which makes high-throughput sorting possible. Both passive and active separation and sorting techniques have been reported in literature. Passive techniques utilize the interaction between particles, flow field and the channel structure and do not require external fields. On the other hand, active techniques make use of external fields in various forms but offer better performance. This paper provides an extensive review of various passive and active separation techniques including basic theories and experimental details. The working principles are explained in detail, and performances of the devices are discussed.     

A portable,hand-powered microfluidic device for sorting of biological particles

Manually hand-powered portable microfluidic devices are cheap alternatives for point-of-care diagnostics. Currently, on-field tests are limited by the use of bulky syringe pumps, pressure controller and equipment. In this work, we present a manually operated microfluidic device incorporated with a groove-based channel. We show that the device is capable to effectively sort particles/cells by manual hand powering. First, the grooved-based channel with differently sized polystyrene particles was characterized using syringe pumps to study their distributions under various flow rate conditions. Afterward, the particle mixtures were sorted manually using hand power to verify the capability of this device. Finally, the manually operated device was used to sort platelets from peripheral blood mononuclear cells (PBMCs). The platelets were collected with a purity of ~ 100%. The purity of PBMCs was enhanced from 0.8 to 10.4% after multiple processes which results in an enrichment ratio of 13.8. During the process of manual hand pumping, the flow fluctuation caused by unstable injection will not influence the sorting performance. Due to its simplicity, this manually operated microfluidic chip is suitable for outfield settings.     

Continuous-flow in-droplet magnetic particle separation in a droplet-based microfluidic platform

This paper presents a continuous-flow in-droplet magnetic particle separation in a droplet-based microfluidic device for magnetic bead-based bioassays. Two functions, electrocoalescence and magnetic particle manipulation, are performed in this device. A pair of charging metallic needles is inserted into two aqueous channels of the device. By electrostatic force, two different solutions can be merged to be mixed at a junction of droplet generation. The manipulation of magnetic particles is achieved using an externally applied magnetic field. The magnetic particles are separated by the magnetic field to one side of the droplet and extracted by splitting the droplet into two daughter droplets: one contains the majority of the magnetic particles and the other is almost devoid of magnetic particles. The applicability of the continuous-flow in-droplet magnetic particle separation is demonstrated by performing a proof-of-concept immunoassay between streptavidin-coated magnetic beads and biotin labelled with fluorescence. This approach will be useful for various biological and chemical analyses and compartmentalization of small samples.     

Sample preconcentration in microfluidic devices

Microfluidic systems have attracted considerable attention and have experienced rapid growth in the past two decades due to advantages associated with miniaturization, integration, and automation. Poor detection sensitivities mainly attributed to the small dimensions of these lab-on-a-chip (LOC) devices; however, sometimes can greatly hinder their practical applications in detecting low-abundance analytes, particularly those in bio-samples. Although off-chip sample pretreatment strategies can be used to address this problem prior to analysis, they may introduce contaminants or lead to an undesirable loss of some original sample volume. Moreover, they are often time-consuming and labor-intensive. Toward the goals of automation, improvement in analytical efficiency, and reductions in sample loss and contamination, many on-chip sample preconcentration techniques based on different working principles for improving the detection sensitivity have been developed and implemented in microchips. The aim of this article is to review recent works in microchip-based sample preconcentration techniques and give detailed discussions about these techniques. We start with a brief introduction regarding the importance of preconcentration techniques in microfluidics and the classification of these techniques based on their concentration mechanisms, followed by in-depth discussions of about these techniques. Finally, personal perspectives on microfluidic-based sample preconcentration will be provided. These advancements in microfluidic sample preconcentration techniques may provide promising strategies for improving the detection sensitivities of LOC devices in many practical applications.     

A facile strategy to integrate robust porous aluminum foil into microfluidic chip for sorting particles

High efficiency integration of functional microdevices into microchips is crucial for broad microfluidic applications. Here, a device-insertion and pressure sealing method was proposed to integrate robust porous aluminum foil into a microchannel for microchip functionalization which demonstrate the advantage of high efficient foil microfabrication and facile integration into the microfluidic chip. The porous aluminum foil with large area (10 × 10 mm²) was realized by one-step femtosecond laser perforating technique within few minutes and its pores size could be precisely controlled from 3 μm to millimeter scale by adjusting the laser pulse energy and pulse number. To verify the versatility and flexibility of this method, two kinds of different microchips were designed and fabricated. The vertical-sieve 3D microfluidic chip can separate silicon dioxide (SiO₂) microspheres of two different sizes (20 and 5 μm), whereas the complex stacking multilayered structures (sandwich-like) microfluidic chip can be used to sort three different kinds of SiO₂ particles (20, 10 and 5 μm) with ultrahigh separation efficiency of more than 92%. Furthermore, these robust filters can be reused via cleaning by backflow (mild clogging) or disassembling (heavy clogging).     

Particle focusing in microfluidic devices

Focusing particles (both biological and synthetic) into a tight stream is usually a necessary step prior to counting, detecting, and sorting them. The various particle focusing approaches in microfluidic devices may be conveniently classified as sheath flow focusing and sheathless focusing. Sheath flow focusers use one or more sheath fluids to pinch the particle suspension and thus focus the suspended particles. Sheathless focusers typically rely on a force to manipulate particles laterally to their equilibrium positions. This force can be either externally applied or internally induced by channel topology. Therefore, the sheathless particle focusing methods may be further classified as active or passive by the nature of the forces involved. The aim of this article is to introduce and discuss the recent developments in both sheath flow and sheathless particle focusing approaches in microfluidic devices.     

Application of polydopamine in biomedical microfluidic devices

Polydopamine (PDA) is a bioinspired material with tremendous potential for applications involving surface modifications. By simply immersing the substrate in the dopamine monomer solution, we are able to apply a hydrophilic and biofunctional PDA coating that adheres strongly to any surface, including (super)hydrophobic surface, with unprecedented ease. Using PDA, almost any materials can be immobilized on the surface in a single step by mixing them with the dopamine monomer solution. This review provides a comprehensive coverage of the applications of PDA in the device fabrication, surface modification, and biofunctionalization of biomedical microfluidic devices. While discussing the advantages and limitations of PDA, we pay special attention to its unique properties that specifically benefit biomedical microfluidic devices. We also discuss other potential applications of PDA beyond the current development. Through this review, we hope to promote PDA and encourage a broader adoption of PDA by the microfluidic community.     

Two-stage polymer embossing of co-planar microfluidic features for microfluidic devices

A two-stage embossing technique for fabricating microchannels for microfluidic devices is presented. A micromachined aluminum mold is used to emboss a polyetherimide (PEI) substrate with a relatively high glass transition temperature (T_g). The embossed PEI is then used as a mold for embossing an amorphous polyethylene terephthalate (APET) substrate with a lower T_g. The resulting APET substrate has the same features as those of the aluminum mold. Successful transfer of features from the aluminum mold to the APET substrate was verified by profilometry, and an application of this method in production of a microfluidic device is presented.     

Micro-injection moulding of polymer microfluidic devices

Microfluidic devices have several applications in different fields, such as chemistry, medicine and biotechnology. Many research activities are currently investigating the manufacturing of integrated microfluidic devices on a mass-production scale with relatively low costs. This is especially important for applications where disposable devices are used for medical analysis. Micromoulding of thermoplastic polymers is a developing process with great potential for producing low-cost microfluidic devices. Among different micromoulding techniques, micro-injection moulding is one of the most promising processes suitable for manufacturing polymeric disposable microfluidic devices. This review paper aims at presenting the main significant developments that have been achieved in different aspects of micro-injection moulding of microfluidic devices. Aspects covered include device design, machine capabilities, mould manufacturing, material selection and process parameters. Problems, challenges and potential areas for research are highlighted.     

Inertial particle focusing and spacing control in microfluidic devices

Focusing particles into a tight stream is usually a necessary step prior to counting, detecting and sorting them. Meanwhile, particle spacing control in microfluidic devices could also be applied in the field of accurate cell detection, material synthesis and chemical reaction. To achieve simultaneous particle focusing and spacing control, a novel microchannel composed by Dean and sheath flow section was proposed and fabricated according to the elaborated design principle with its manipulating performance in situ visualized. Using microspheres with a few microns as a template, the trajectory of the particles was discovered to follow lateral migration and reach certain equilibrium positions at the end of the designed Dean section. After being focused, the streamline was further concentrated and centralized with a controllable interparticle distance in sheath flow section. For sheath flow section, the angle between symmetrical tributaries and the mainstream channel and abrupt constriction/expansion structure of mainstream channel as important channel geometric features were investigated to minimize the focusing streamline width and optimize spacing control. An modified analytical model for sheath flow with different tributary angles was derived and proved to well describe the microsphere spacing control process.     

Methods for counting particles in microfluidic applications

Microfluidic particle counters are important tools in biomedical diagnostic applications such as flow cytometry analysis. Major methods of counting particles in microfluidic devices are reviewed in this paper. The microfluidic resistive pulse sensor advances in sensitivity over the traditional Coulter counter by improving signal amplification and noise reduction techniques. Nanopore-based methods are used for single DNA molecule analysis and the capacitance counter is useful in liquids of low electrical conductivity and in sensing the changes of cell contents. Light-scattering and light-blocking counters are better for detecting larger particles or concentrated particles. Methods of using fluorescence detection have the capability for differentiating particles of similar sizes but different types that are labeled with different fluorescent dyes. The micro particle image velocimetry method has also been used for detecting and analyzing particles in a flow field. The general limitation of microfluidic particle counters is the low throughput which needs to be improved in the future. The integration of two or more existing microfluidic particle counting techniques is required for many practical on-chip applications.     

Positioning system for particles in microfluidic structures

Fast continuous flow detection of biomolecules in lab-on-a-chip structures is a challenging task. Combining these molecules with small magnetic particles, the interaction between their stray field and, e.g., magneto-resistive sensors can be used to indirectly prove the biomolecules. To position the particles on top of a sensor array at the bottom of the flow channel, we propose a microfluidic structure of changing channel height combining hydrodynamic and gravitational effects. We present numerical calculations predicting an increase in the capture rate by more than 100% in comparison to a straight channel. We experimentally realize an optical analysis of the specific binding of biotin-functionalized Chemagen beads on a streptavidin-coated surface. To prove the binding is not due to the surface effects, a second uncoated bead species is employed.     

Driving and sorting of the fluorescent droplets on digital microfluidic platform

In this paper, we present a digital microfluidic droplet sorting platform to achieve automated droplet sorting based on fluorescent detection. We design and fabricate a kind of digital microfluidic chip for manipulating nano-liter-sized liquid droplets, and the chip is integrated with a fluorescence-initiated feedback system for real-time sorting control. The driving and sorting characteristics of fluorescent droplets encapsulating fluorescent-labeled particles are studied on this platform. The droplets dispensed from on-chip reservoir electrode are transported to a fluorescence detection site and sorted according to their fluorescence signals. The fluorescent droplets and non-fluorescent droplets are successfully separated and the number of fluorescent particles inside each droplet is quantified by its fluorescent intensity. We realize droplet sorting at 20 Hz and obtain a linear relationship between the fluorescent particle concentrations and the fluorescence signals. This work is easily adapted for sorting out fluorescent-labeled microparticles, cells and bacteria and thus has the potential of quantifying catalytic or regulatory bio-activities.     

Rapid prototyping of shrinkable BOPS-based microfluidic devices

Biaxially oriented polystyrene (BOPS) is a commercialized packaging material, which has the advantages of biocompatibility, non-toxic, transparency, light-weight and cost-effective. Due to the stress accumulated from both directions in plane during the fabrication process, when BOPS was reheated above the glass transition temperature, an isotropic shrinkage will occur. This study proposed a low-cost and rapid prototyping method for the fabrication of BOPS-based microfluidics device. Both laser ablation and micro-milling were used for the fabrication of microchannels on the surface of the BOPS sheet, after thermal induced shrinkage, microchannels with finer microstructure could be achieved. For the sealing of fabricated microchannels on BOPS, two approaches were made using a layer of BOPS or a layer of polyester adhesive film. The thermal induced shrinkage and bonding strength were carefully studied in this study. Several microfluidic devices, including a droplet generator and a diffusion mixer were also fabricated for demonstration. The proposed fabrication method for BOPS-based microfluidics is simple, rapid, cost-effective and without the requirement of cleanroom facility, with help of thermal induced shrinkage, finer structure with high resolution could be achieved with conventional lab tools.     

Numerical modeling of electrowetting processes in digital microfluidic devices

We use a three-dimensional multiphase lattice-Boltzmann model to study basic operations such as transport, merging and splitting of nanoliter water droplets actuated by electrowetting in digital microfluidic devices. In a first step, numerical and analytical predictions for the droplet transport velocity are compared and very good agreement is obtained for a wide range of contact angles. The same algorithm is employed then to study the dynamics of the splitting processes at different contact angles and different geometries of the cell. The configuration of the liquid droplet involved in a splitting process and the dependence of the splitting time on the transport velocity are also investigated and phenomenological laws describing these processes are also proposed.     

Safe and cost-effective rapid-prototyping of multilayer PMMA microfluidic devices

The rapid translation of research from bench to bedside, as well as the generation of commercial impact, has never been more important for both academic and industrial researchers. It is therefore unsurprising that more and more microfluidic groups are investigating research using a wide range of thermoplastics which can be readily translated to large-scale manufacturing, if the technology is taken to commercialisation. While structuring, via additive or subtractive manufacturing, is becoming relatively easy through the use of commercial-grade devices, reliable and fast assembly remains a challenge. In this article, we propose an innocuous and cost-effective, under 2-min technique which enables the bonding of multiple poly(methyl methacrylate) layers. This bonding technique relies on the application of small amounts (10 µl/cm²) of ethanol, low temperatures (70 °C) and relatively low pressures (~1.6 MPa). Our characterisation analysis shows that using a bonding time of 2 min is enough to produce a strong bond able to withstand pressures always above 6.2 MPa (with mean of 8 MPa, highest reported in the literature), with minimal channel deformation (<5%). This technique, which we demonstrate on assembly comprising up to 19 layers, presents an exciting improvement in the field of rapid prototyping of microfluidic devices, enabling extremely fast design cycles in microfluidic research with a material compatible with mass manufacturing, thus allowing a smoother transition from the laboratory to the market. Beyond the research community, this robust prototyping technique has the potential to impact on the deliverability of other scientific endeavours including educational projects and will directly fuel the fluidic maker movement.     

Automation of three-dimensional cell culture in arrayed microfluidic devices

The increasing interest in studying the interactions between cells and the extracellular matrix (ECM) has created a need for high throughput low-cost three-dimensional (3D) culture systems. The recent development of tubeless microfluidics via passive pumping provides a high throughput microchannel culture platform compatible with existing high throughput infrastructures (e.g., automated liquid handlers). Here, we build on a previously reported high throughput two-dimensional system to create a robust automated system for 3D culture. Operational controls including temperature and sample handling have been characterized and automated. Human mammary fibroblasts (HMFs) suspended in type I collagen are loaded and cultured in microchannel arrays and used to optimize the system operational parameters. A Peltier cooler maintains the collagen as a liquid at 4 °C during cell seeding, followed by polymerization at 37 °C. Optimization of this platform is discussed (e.g., controlling collagen contraction, increasing cell viability, preventing the removal of microchannel contents), and 3D distribution of HMFs is examined by fluorescent microscopy. Finally, we validate the platform by automating a previously developed 3D breast carcinoma coculture assay. The platform allows more efficient 3D culture experiments and lays the foundation for high throughput studies of cell-ECM interactions.     

Modular component design for portable microfluidic devices

A new modular design concept for microfluidic devices is proposed and demonstrated in this study. We designed three key modular microfluidic components: pumps, valves, and reservoirs, and demonstrated that a microfluidic device with specific functions can be easily assembled with those key modular components. Our pumps are man-powerable so that the assembled microfluidic devices require no any other power sources like expensive syringe pumps or air compressors. This feature makes the assembled microfluidic devices completely portable. We also combined our assembled device with other existing mixing microchannels to serve as the mixing and loading system in polymerase chain reaction experiment to amplify DNA successfully. This result shows that those modular components can be integrated into other microchannels, implying great potential applications of the modular design.     

Novel continuous particle sorting in microfluidic chip utilizing cascaded squeeze effect

This article presents a novel technique for the continuous sorting and collection of microparticles in a microfluidic chip using a cascaded squeeze effect. In the proposed approach, microparticles of different sizes are separated from the sample stream using sheath flows and are then directed to specific side channels for collection. The sheath flows required to separate the particles are generated using a single high voltage supply integrated with a series of variable resistors designed to create electric fields of different intensities at different points of the microchip. Numerical simulations are performed to analyze the electrical potential contours and flow streamlines within the microchannel. Experimental trials show that the microchip is capable of continuously separating microparticles with diameters of 5, 10 and 20 μm, respectively. To further evaluate the performance of the microchip, a sample composed of yeast cells and polystyrene beads is sorted and collected. The results indicate that the microchip achieves a recovery ratio of 87.7% and a yield ratio of 94.1% for the yeast cells and therefore attains a comparable performance to that of a large-scale commercial flow cytometer. Importantly, the high performance of the microchip is achieved without the need for a complex control system or for sophisticated actuation mechanisms such as embedded microelectrodes, ultrasonic generators, or micropumps, and so forth.