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.

Sub-micrometer-precision, three-dimensional (3D) hydrodynamic focusing via "microfluidic drifting".

In this article, we demonstrate single-layered, "microfluidic drifting" based three-dimensional (3D) hydrodynamic focusing devices with particle/cell focal positioning approaching submicron precision along both lateral and vertical directions. By systematically optimizing channel geometries and sample/sheath flow rates, a series of "microfluidic drifting" based 3D hydrodynamic focusing devices with different curvature angles are designed and fabricated. Their performances are then evaluated using confocal microscopy, fast camera imaging, and side-view imaging techniques. Using a device with a curvature angle of 180°, we have achieved a standard deviation of ±0.45 µm in particle focal position and a coefficient of variation (CV) of 2.37% in flow cytometric measurements. To the best of our knowledge, this is the best CV that has been achieved using a microfluidic flow cytometry device. Moreover, the device showed the capability to distinguish 8 peaks when subjected to a stringent 8-peak rainbow calibration test, signifying the ability to perform sensitive, accurate tests similar to commercial flow cytometers. We have further tested and validated our device by detection of HEK-293 cells. With its advantages in simple fabrication (i.e., single-layered device), precise 3D hydrodynamic focusing (i.e., submicrometer precision along both lateral and vertical directions), and high detection resolution (i.e., low CV), our method could serve as an important basis for high-performance, mass-producible microfluidic flow cytometry.

Surface acoustic wave microfluidics.

The recent introduction of surface acoustic wave (SAW) technology onto lab-on-a-chip platforms has opened a new frontier in microfluidics. The advantages provided by such SAW microfluidics are numerous: simple fabrication, high biocompatibility, fast fluid actuation, versatility, compact and inexpensive devices and accessories, contact-free particle manipulation, and compatibility with other microfluidic components. We believe that these advantages enable SAW microfluidics to play a significant role in a variety of applications in biology, chemistry, engineering and medicine. In this review article, we discuss the theory underpinning SAWs and their interactions with particles and the contacting fluids in which they are suspended. We then review the SAW-enabled microfluidic devices demonstrated to date, starting with devices that accomplish fluid mixing and transport through the use of travelling SAW; we follow that by reviewing the more recent innovations achieved with standing SAW that enable such actions as particle/cell focusing, sorting and patterning. Finally, we look forward and appraise where the discipline of SAW microfluidics could go next.

An on-chip, multichannel droplet sorter using standing surface acoustic waves.

The emerging field of droplet microfluidics requires effective on-chip handling and sorting of droplets. In this work, we demonstrate a microfluidic device that is capable of sorting picoliter water-in-oil droplets into multiple outputs using standing surface acoustic waves (SSAW). This device integrates a single-layer microfluidic channel with interdigital transducers (IDTs) to achieve on-chip droplet generation and sorting. Within the SSAW field, water-in-oil droplets experience an acoustic radiation force and are pushed toward the acoustic pressure node. As a result, by tuning the frequency of the SSAW excitation, the position of the pressure nodes can be changed and droplets can be sorted to different outlets at rates up to 222 droplets s(-1). With its advantages in simplicity, controllability, versatility, noninvasiveness, and capability to be integrated with other on-chip components such as droplet manipulation and optical detection units, the technique presented here could be valuable for the development of droplet-based micro total analysis systems (µTAS).

Holographically formed, acoustically switchable gratings based on polymer-dispersed liquid crystals.

We report holographic polymer-dispersed liquid crystal (H-PDLC) gratings driven by surface acoustic waves (SAWs). Our experiments show that upon applying SAWs, the H-PDLC grating exhibited switchable properties: The diffraction of the H-PDLC grating decreased, whereas the transmission increased. This acoustically switchable behavior is due to the acoustic streaming-induced realignment of liquid crystals as well as absorption-resulted thermal diffusion. Such SAW-driven H-PDLC gratings are potentially useful in many photonic applications, such as optical switches, spatial light modulators, and switchable add/drop filters.

Optofluidic imaging: now and beyond.

More than a decade of research work in optofluidics has yielded a large catalogue of optofluidic elements that can manipulate light at the micro-scale (e.g., lenses, prisms). Although these elements have proven useful for many on-chip processes (e.g., miniaturized flow cytometry, interferometry and sample spectroscopy), certain deficiencies have precluded their use in micro-scale imaging. However, recent work in optofluidic imaging has avoided optofluidic elements entirely and focused instead on image capture and composition techniques, demonstrating impressive resolution in both 2D imagery and 3D tomography. In this Focus article, we will discuss some of the recent successes in optofluidic imaging and will expound our expectations for the near future of the optofluidic imaging discipline.

Tunable, pulsatile chemical gradient generation via acoustically driven oscillating bubbles.

We present a novel concept of generating both static and pulsatile chemical gradients using acoustically activated bubbles designed in a ladder-like arrangement. Furthermore, by regulating the amplitude of the bubble oscillation, we demonstrate that the chemical gradient profiles can be effectively tuned.

A droplet-based, optofluidic device for high-throughput, quantitative bioanalysis.

Analysis of chemical or biomolecular contents in a tiny amount of specimen presents a significant challenge in many biochemical studies and diagnostic applications. In this work, we present a single-layer, optofluidic device for real-time, high-throughput, quantitative analysis of droplet contents. Our device integrates an optical fiber-based, on-chip detection unit with a droplet-based microfluidic unit. It can quantitatively analyze the contents of individual droplets in real-time. It also achieves a detection throughput of 2000 droplets per second, a detection limit of 20 nM, and an excellent reproducibility in its detection results. In a proof-of-concept study, we demonstrate that our device can be used to perform detection of DNA and its mutations by monitoring the fluorescent signal changes of the target DNA/molecular beacon complex in single droplets. Our approach can be immediately extended to a real-time, high-throughput detection of other biomolecules (such as proteins and viruses) in droplets. With its advantages in throughput, functionality, cost, size, and reliability, the droplet-based optofluidic device presented here can be a valuable tool for many medical diagnostic applications.

Standing surface acoustic wave (SSAW) based multichannel cell sorting.

We introduce a novel microfluidic device for cell sorting in continuous flow using tunable standing surface acoustic waves. This method allows individual cells to be precisely directed into five different outlet channels in a single step. It is versatile, simple, label-free, non-invasive, and highly controllable.

Single-shot characterization of enzymatic reaction constants Km and kcat by an acoustic-driven, bubble-based fast micromixer.

In this work we present an acoustofluidic approach for rapid, single-shot characterization of enzymatic reaction constants K(m) and k(cat). The acoustofluidic design involves a bubble anchored in a horseshoe structure which can be stimulated by a piezoelectric transducer to generate vortices in the fluid. The enzyme and substrate can thus be mixed rapidly, within 100 ms, by the vortices to yield the product. Enzymatic reaction constants K(m) and k(cat) can then be obtained from the reaction rate curves for different concentrations of substrate while holding the enzyme concentration constant. We studied the enzymatic reaction for ß-galactosidase and its substrate (resorufin-ß-D-galactopyranoside) and found K(m) and k(cat) to be 333 ± 130 µM and 64 ± 8 s(-1), respectively, which are in agreement with published data. Our approach is valuable for studying the kinetics of high-speed enzymatic reactions and other chemical reactions.

Design of acoustic beam aperture modifier using gradient-index phononic crystals.

This article reports the design concept of a novel acoustic beam aperture modifier using butt-jointed gradient-index phononic crystals (GRIN PCs) consisting of steel cylinders embedded in a homogeneous epoxy background. By gradually tuning the period of a GRIN PC, the propagating direction of acoustic waves can be continuously bent to follow a sinusoidal trajectory in the structure. The aperture of an acoustic beam can therefore be shrunk or expanded through change of the gradient refractive index profiles of the butt-jointed GRIN PCs. Our computational results elucidate the effectiveness of the proposed acoustic beam aperture modifier. Such an acoustic device can be fabricated through a simple process and will be valuable in applications, such as biomedical imaging and surgery, nondestructive evaluation, communication, and acoustic absorbers.

Surface acoustic wave (SAW) acoustophoresis: now and beyond.

On-chip manipulation of micro-objects has long been sought to facilitate fundamental biological studies and point-of-care diagnostic systems. In recent years, research on surface acoustic wave (SAW) based micro-object manipulation (i.e., SAW acoustophoresis) has gained significant momentum due to its many advantages, such as non-invasiveness, versatility, simple fabrication, easy operation, and convenient integration with other on-chip units. SAW acoustophoresis is especially useful for lab-on-a-chip applications where a compact and non-invasive biomanipulation technique is highly desired. In this Focus article, we discuss recent advancements in SAW acoustophoresis and provide some perspectives on the future development of this dynamic field.

Tunable patterning of microparticles and cells using standing surface acoustic waves.

We have developed an acoustic-based tunable patterning technique by which microparticles or cells can be arranged into reconfigurable patterns in microfluidic channels. In our approach, we use pairs of slanted-finger interdigital transducers (SFITs) to generate a tunable standing surface acoustic wave field, which in turn patterns microparticles or cells in one- or two-dimensional arrays inside the microfluidic channels--all without the assistance of fluidic flow. By tuning the frequency of the input signal applied to the SFITs, we have shown that the cell pattern can be controlled with tunability of up to 72%. This acoustic-based tunable patterning technique has the advantages of wide tunability, non-invasiveness, and ease of integration to lab-on-a-chip systems, and shall be valuable in many biological and colloidal studies.

On-chip manipulation of single microparticles, cells, and organisms using surface acoustic waves.

Techniques that can dexterously manipulate single particles, cells, and organisms are invaluable for many applications in biology, chemistry, engineering, and physics. Here, we demonstrate standing surface acoustic wave based "acoustic tweezers" that can trap and manipulate single microparticles, cells, and entire organisms (i.e., Caenorhabditis elegans) in a single-layer microfluidic chip. Our acoustic tweezers utilize the wide resonance band of chirped interdigital transducers to achieve real-time control of a standing surface acoustic wave field, which enables flexible manipulation of most known microparticles. The power density required by our acoustic device is significantly lower than its optical counterparts (10,000,000 times less than optical tweezers and 100 times less than optoelectronic tweezers), which renders the technique more biocompatible and amenable to miniaturization. Cell-viability tests were conducted to verify the tweezers' compatibility with biological objects. With its advantages in biocompatibility, miniaturization, and versatility, the acoustic tweezers presented here will become a powerful tool for many disciplines of science and engineering.

An integrated, multiparametric flow cytometry chip using "microfluidic drifting" based three-dimensional hydrodynamic focusing.

In this work, we demonstrate an integrated, single-layer, miniature flow cytometry device that is capable of multi-parametric particle analysis. The device integrates both particle focusing and detection components on-chip, including a "microfluidic drifting" based three-dimensional (3D) hydrodynamic focusing component and a series of optical fibers integrated into the microfluidic architecture to facilitate on-chip detection. With this design, multiple optical signals (i.e., forward scatter, side scatter, and fluorescence) from individual particles can be simultaneously detected. Experimental results indicate that the performance of our flow cytometry chip is comparable to its bulky, expensive desktop counterpart. The integration of on-chip 3D particle focusing with on-chip multi-parametric optical detection in a single-layer, mass-producible microfluidic device presents a major step towards low-cost flow cytometry chips for point-of-care clinical diagnostics.

Three-dimensional continuous particle focusing in a microfluidic channel via standing surface acoustic waves (SSAW).

Three-dimensional (3D) continuous microparticle focusing has been achieved in a single-layer polydimethylsiloxane (PDMS) microfluidic channel using a standing surface acoustic wave (SSAW). The SSAW was generated by the interference of two identical surface acoustic waves (SAWs) created by two parallel interdigital transducers (IDTs) on a piezoelectric substrate with a microchannel precisely bonded between them. To understand the working principle of the SSAW-based 3D focusing and investigate the position of the focal point, we computed longitudinal waves, generated by the SAWs and radiated into the fluid media from opposite sides of the microchannel, and the resultant pressure and velocity fields due to the interference and reflection of the longitudinal waves. Simulation results predict the existence of a focusing point which is in good agreement with our experimental observations. Compared with other 3D focusing techniques, this method is non-invasive, robust, energy-efficient, easy to implement, and applicable to nearly all types of microparticles.

Beam bending via plasmonic lenses.

We have designed and characterized three different types of plasmonic lenses that cannot only focus, but can also bend electromagnetic (EM) waves. The bending effect is achieved by constructing an asymmetric phase front caused by varying phase retardations in EM waves as they pass through a plasmonic lens. With an incident wave normal to the lens surface, light bends up to 8° off the axial direction. The optical wave propagation was numerically investigated using the finite-difference time-domain (FDTD) method. Simulation results show that the proposed plasmonic lenses allow effective beam bending under both normal and tilted incidence. With their relatively large bending range and capability to perform in the far field, the plamsonic lenses described in this article could be valuable in applications such as photonic communication and plasmonic circuits.

Tunable two-dimensional liquid gradient refractive index (L-GRIN) lens for variable light focusing.

We report a two-dimensional (2D) tunable liquid gradient refractive index (L-GRIN) lens for variable focusing of light in the out-of-plane direction. This lens focuses a light beam through a liquid medium with a 2D hyperbolic secant (HS) refractive index gradient. The refractive index gradient is established in a microfluidic chamber through the diffusion between two fluids with different refractive indices, i.e. CaCl(2) solution and deionized (DI) water. The 2D HS refractive index profile and subsequently the focal length of the L-GRIN lens can be tuned by changing the ratio of the flow rates of the CaCl(2) solution and DI water. The focusing effect is experimentally characterized through side-view and top-view image analysis, and the experimental data match well with the results from ray-tracing optical simulations. Advantages of the 2D L-GRIN lens include simple device fabrication procedure, low fluid consumption rate, convenient lens-tuning mechanism, and compatibility with existing microfluidic devices. We expect that with further optimizations, this 2D L-GRIN lens can be used in many optics-based lab-on-a-chip applications.

Optofluidic tunable microlens by manipulating the liquid meniscus using a flared microfluidic structure.

We have designed, demonstrated, and characterized a simple, novel in-plane tunable optofluidic microlens. The microlens is realized by utilizing the interface properties between two different fluids: CaCl(2)solution and air. A constant contact angle of ∼90° is the pivotal factor resulting in the outward bowing and convex shape of the CaCl(2) solution-air interface. The contact angle at the CaCl(2) solution-air interface is maintained by a flared structure in the polydimethylsiloxane channel. The resulting bowing interface, coupled with the refractive index difference between the two fluids, results in effective in-plane focusing. The versatility of such a design is confirmed by characterizing the intensity of a traced beam experimentally and comparing the observed focal points with those obtained via ray-tracing simulations. With the radius of curvature conveniently controlled via fluid injection, the resulting microlens has a readily tunable focal length. This ease of operation, outstandingly low fluid usage, large range tunable focal length, and in-plane focusing ability make this lens suitable for many potential lab-on-a-chip applications such as particle manipulation, flow cytometry, and in-plane optical trapping.