Acoustofluidics 9: Modelling and applications of planar resonant devices for acoustic particle manipulation

This article introduces the design, construction and applications of planar resonant devices for particle and cell manipulation. These systems rely on the pistonic action of a piezoelectric layer to generate a one dimensional axial variation in acoustic pressure through a system of acoustically tuned layers. The resulting acoustic standing wave is dominated by planar variations in pressure causing particles to migrate to planar pressure nodes (or antinodes depending on particle and fluid properties). The consequences of lateral variations in the fields are discussed, and rules for designing resonators with high energy density within the appropriate layer for a given drive voltage presente

Membrane Deflection-based Fabrication and Design Automation for Integrated Acoustofluidics

Continuous-flow microfluidic large-scale integration (mLSI) is a developing field first introduced in the early 2000s, that continues to offer promising solutions to many biochemical, biophysical and biomedical problems. In his seminal paper, Thorsen et al. 2002 demonstrated the fabrication of high-density microfluidic systems capable of complex fluidic routing in combinatory arrays of multiplexors, mixers, and storage assemblies integrated with micromechanical valves. mLSI has been a powerful tool for scientific research by allowing for dramatic reduction in the volume of reagent needed for experimentation and offering highly parallelizable and dynamic process flows. These systems have since been the focus of strong interdisciplinary academic research efforts. Despite the success in scientific applications, the mLSI technologies have not found widespread use in commercial environments. One critical issue preventing mLSI to realize its full potential is the need for specialized fabrication techniques that are scalable and more suitable for the unique requirements of biology. The work presented here demonstrates an mLSI integrated acoustofluidic platform that offers versatility while maintaining a robust fabrication process. In particular, conductive liquid metal-based acoustic transducers integrated with micromechanical valves to facilitate dynamic switching of the resonant frequency of the device and generated surface acoustic waves (SAWs) is demonstrated. Shortcomings in the fabrication of fluidic channels for mLSI integrated acoustofluidic applications are examined, and solutions to these problems are presented. A novel and scalable soft-lithographic method is introduced, that allows for the fabrication of large valvable channels with tunable height that exceeds practical limitations dictated by previous photolithographic techniques. A thorough characterization of this method and demonstration of its robustness are included here as a promising data to promote further exploration of the technique as a viable commercial solution for the fabrication of many classes of mLSI bio-devices. The testing of a computeraided design software, Columba, is briefly discussed

IN-LINE MICROFLUIDIC PARTICLE PRECONCENTRATOR AND DETECTOR FOR CONTINUOUS FLOW MONITORING

This dissertation presents the design and prototyping of three in-line microfluidic devices for continuous monitoring of particulate flows. The three devices are AC electrokinetic (ACEK) and acoustic sample preconcentration techniques for resettable particle enrichment, and an in-line somatic cell counter for mastitis monitoring. For the ACEK preconcentrator, ACEK is a new and promising technique to manipulate micro/bio-fluid and particles. There are many advantages over other techniques, such as low applied voltage, low cost, portability and notable biocompatibility of lab-on-a-chip (LOC) device. We successfully developed a 3D multi-level electrode platform to extract bioparticles via AC electroosmosis (ACEO) and negative Dielectrophoresis (DEP). Based on ACEO and N-DEP, the device can exert a drag force on particles through fluid motion and collect and concentrate particles. Optimization with respect to AC frequency, external pumping rate and opening size of mesh electrode have been performed. This research also studies the concentration effect by acoustic wave on diatom cells in seawater environment, since ACEK has limitation in high conductivity medium. Acoustic trapping uses mechanical resonance to focus the target particles into the designated trapping area. It has the advantages of high trapping efficiency, contactless trapping and compatibility with various fluids. Furthermore, since the trapping effect and the vertical trapping location are dependent on the particle properties, binary particle separation and sorting are also highly possible. Another contribution of this dissertation is the ACEK based capacitive somatic cell counter for use in dairy industry. Using our design, capacitive sensing is capable of detecting and quantifying target concentration in many types of biological solutions. The capacitance changing rate of device can be correlated with different concentrations of somatic cells. In this work, we successfully detected the concentration level of somatic cells in raw milk. The results were verified by flow cytometry

Particle Enrichment in Longitudinal Standing Bulk Acoustic Wave Microfluidics

Separation, isolation, and enrichment of targeted nano- and microparticles are critical to a variety of biomedical applications from clinical research (development of therapeutics and diagnostics) to fundamental investigations that require concentration of specific cells from culture, separation of target species from heterogenous mixtures, or controlled perturbation of cells and microorganisms to determine their response to stimuli. Numerous techniques are available for bench-scale and medical settings; however, these traditional approaches are often labor intensive, time-consuming, costly, and/or require modification of the target. Efficiency and specificity are also lacking. Recently, techniques that exploit the similar scales of microfluidic technologies and the intrinsic properties of cells have allowed for increased automation, reduced reagent waste, and decreased cost, as well as improved performance. So-called lab-on-a-chip (LOC) approaches enable rapid fabrication and optimization of small-scale, low-volume microchannels capable of high performance enrichment and separation owing to precise control of the forces driving the manipulation. Depending on the physics underlying a particular method, devices are classified as optical, hydrodynamic, dielectrophoretic, magnetic, or acoustic. Acoustics, and specifically ultrasound, permits noncontact cell separation and retention, which reduces the potential for undesirable surface interactions and physical stress on sensitive biological samples. Typically, separation is achieved by pinning a standing wave perpendicular (conventional lateral acoustophoresis) or parallel (longitudinal acoustic trapping) to the direction of flow. In this thesis, we report a novel longitudinal standing bulk acoustic wave (LSBAW) microfluidic channel that incorporates pairs of pillar arrays oriented perpendicular to the inflow direction. The pillar arrays act as ‘pseudo walls’ that locally amplify the pressure in the enrichment zone, which can be tuned to overcome the drag force for particles of size greater than a critical diameter. Thus, these particles are preferentially retained within the nodes of the local pressure field. In our study, model predictions were used to guide experimental trapping of particles in microchannels with two pillar configurations. We created six different microfluidic channels with varying inlet/outlet geometries, widths, and pillar shapes. Model results showed pressure field amplification caused by the ‘pseudo walls’ bounding the enrichment zone of each design. We also demonstrated trapping of polystyrene beads (5 μm and 20 μm) and 10 μm fluorescent hollow glass spheres during actuation at various predicted half-wave resonances of these devices. Certain channel architectures achieved acoustic field amplification suitable for particle trapping at flow rates up to ~20 μL/min. In addition, the simulated pressure fields (eigenmodes) were consistent with experimentally observed mode shapes, which validated our modeling approach. Computational and experimental results suggest that LSBAW pillar geometries and flow parameters can be tuned to achieve enhanced enrichment of targeted particles in a predefined region

Recent advances in non-optical microfluidic platforms for bioparticle detection

The effective analysis of the basic structure and functional information of bioparticles are of great significance for the early diagnosis of diseases. The synergism between microfluidics and particle manipulation/detection technologies offers enhanced system integration capability and test accuracy for the detection of various bioparticles. Most microfluidic detection platforms are based on optical strategies such as fluorescence, absorbance, and image recognition. Although optical microfluidic platforms have proven their capabilities in the practical clinical detection of bioparticles, shortcomings such as expensive components and whole bulky devices have limited their practicality in the development of point-of-care testing (POCT) systems to be used in remote and underdeveloped areas. Therefore, there is an urgent need to develop cost-effective non-optical microfluidic platforms for bioparticle detection that can act as alternatives to optical counterparts. In this review, we first briefly summarise passive and active methods for bioparticle manipulation in microfluidics. Then, we survey the latest progress in non-optical microfluidic strategies based on electrical, magnetic, and acoustic techniques for bioparticle detection. Finally, a perspective is offered, clarifying challenges faced by current non-optical platforms in developing practical POCT devices and clinical applications.</p

Cell separation using tilted-angle standing surface acoustic waves

Separation of cells is a critical process for studying cell properties, disease diagnostics, and therapeutics. Cell sorting by acoustic waves offers a means to separate cells on the basis of their size and physical properties in a label-free, contactless, and biocompatible manner. The separation sensitivity and efficiency of currently available acoustic-based approaches, however, are limited, thereby restricting their widespread application in research and health diagnostics. In this work, we introduce a unique configuration of tilted-angle standing surface acoustic waves (taSSAW), which are oriented at an optimally designed inclination to the flow direction in the microfluidic channel. We demonstrate that this design significantly improves the efficiency and sensitivity of acoustic separation techniques. To optimize our device design, we carried out systematic simulations of cell trajectories, matching closely with experimental results. Using numerically optimized design of taSSAW, we successfully separated 2- and 10-µm-diameter polystyrene beads with a separation efficiency of ~99%, and separated 7.3- and 9.9-µm-polystyrene beads with an efficiency of ~97%. We illustrate that taSSAW is capable of effectively separating particles–cells of approximately the same size and density but different compressibility. Finally, we demonstrate the effectiveness of the present technique for biological–biomedical applications by sorting MCF-7 human breast cancer cells from nonmalignant leukocytes, while preserving the integrity of the separated cells. The method introduced here thus offers a unique route for separating circulating tumor cells, and for label-free cell separation with potential applications in biological research, disease diagnostics, and clinical practice.National Institutes of Health (U.S.) (Grant U01HL114476)National Institutes of Health (U.S.) (New Innovator Award 1DP2OD007209-01)National Science Foundation (U.S.). Materials Research Science and Engineering Centers (Program) (Grant DMR-0820404

Label-free cell separation and sorting in microfluidic systems

Cell separation and sorting are essential steps in cell biology research and in many diagnostic and therapeutic methods. Recently, there has been interest in methods which avoid the use of biochemical labels; numerous intrinsic biomarkers have been explored to identify cells including size, electrical polarizability, and hydrodynamic properties. This review highlights microfluidic techniques used for label-free discrimination and fractionation of cell populations. Microfluidic systems have been adopted to precisely handle single cells and interface with other tools for biochemical analysis. We analyzed many of these techniques, detailing their mode of separation, while concentrating on recent developments and evaluating their prospects for application. Furthermore, this was done from a perspective where inertial effects are considered important and general performance metrics were proposed which would ease comparison of reported technologies. Lastly, we assess the current state of these technologies and suggest directions which may make them more accessible

Microscale methods to investigate and manipulate multispecies biological systems

The continuing threats from viral infectious diseases highlight the need for new tools to study viral interactions with host cells. Understanding how these viruses interact and respond to their environment can help predict outbreaks, shed insight on the most likely strains to emerge, and determine which viruses have the potential to cause significant human illness. Animal studies provide a wealth of information, but the interpretation of results is confounded by the large number of uncontrolled or unknown variables in complex living systems. In contrast, traditional tissue culture approaches have provided investigators a valuable platform with a high degree of experimental control and flexibility, but the static nature of flask-based cell culture makes it difficult to study viral evolution. Serial passaging introduces un-physiological perturbations to cell and virus populations by drastically reducing the number of species with each passage. Low copy, high fitness viral variants maybe eliminated, while in vivo these variants would be essential in determining the virus’ evolutionary fate. Bridging technologies are urgently needed to mitigate the unrealistic dynamics in static flask-based cultures, and the complexity and expense of in vivo experiments. This thesis details the development of a continuous perfusion platform capable of more closely mimicking in vivo cell-virus dynamics, while surpassing the experimental control and flexibility of standard cell culture. First, a microfluidic flow through acoustic device is optimized to enable efficient and controllable separation of cells and viruses. Repeatable isolation of cell and virus species is demonstrated with both a well-characterized virus, Dengue Virus (DENV), and the novel Golden Gate Virus. Next, a platform is built around this device to enable controllable, automated, continuous cell culture. Beads are used to assess system performance and optimize operation. Subsequently, the platform is used to culture both murine hybridoma (4G2) and human monocyte (THP-1) cell lines for over one month, and demonstrate the ability to manipulate population dynamics. Finally, we use the platform to establish a multispecies culture with THP-1 cells and Sindbis Virus (SINV). This work integrates distinct engineering feats to create a platform capable of enhancing existing cell virus studies and opening the door to a variety of high-impact investigations

Acoustic Standing Wave Manipulation of Particles and Cells in Microfluidic Chips

The rise of MEMS and µTAS techniques has created a whole new family of microfluidic devices for a wide range of chemical and biomedical analyses to be performed on small Lab-on-a-chip platforms. The operations often include small samples of particle or cell suspensions on which separation, mixing, trapping or sorting is performed. External fields and forces are used for these operations, and this thesis is specifically focused the development of ultrasonic standing wave technology and the use of acoustic force fields to perform bioanalytical unit operations. The combination of acoustic standing waves and the laminar flow in microfluidics has proven to be well suited for performing particle and cell separation. The fundamental acoustic separator used in this thesis consists of a microfluidic flow channel with a three way flow splitter (trifurcation) in the end of the channel. An acoustic standing wave field is applied to the main flow channel by attaching the transducer underneath the chip. The acoustic standing wave is however obtained perpendicular to the axial propagation of the wave field and the direction of the flow. The half wavelength resonance affects rigid particles or cells driving them into the acoustic pressure node while liquid spheres having other density and compressibility properties may move to the pressure antinode. This enables acoustic separation of different particle types. Blood has proven to be very suitable for acoustic cell manipulation. An application where lipid particles can be removed acoustically from shed blood from open heart surgery is demonstrated. An application for acoustic plasmapheresis is also shown where high quality blood plasma is generated. Different separator designs, device material, and the influence of the separation channel cross-section design are also investigated