David J. Collins

Two-dimensional single-cell patterning with one cell per well driven by surface acoustic waves

In single-cell analysis, cellular activity and parameters are assayed on an individual, rather than population-average basis. Essential to observing the activity of these cells over time is the ability to trap, pattern and retain them, for which previous single-cell-patterning work has principally made use of mechanical methods. While successful as a long-term cell-patterning strategy, these devices remain essentially single use. Here we introduce a new method for the patterning of multiple spatially separated single particles and cells using high-frequency acoustic fields with one cell per acoustic well. We characterize and demonstrate patterning for both a range of particle sizes and the capture and patterning of cells, including human lymphocytes and red blood cells infected by the malarial parasite Plasmodium falciparum. This ability is made possible by a hitherto unexplored regime where the acoustic wavelength is on the same order as the cell dimensions.

Surface acoustic waves for on-demand production of picoliter droplets and particle encapsulation.

Microscopic water-in-oil droplets are a versatile chemical and biological platform whose dimensions result in short reaction times and require minuscule amounts of reagent. Methods exist for the production of droplets, though the vast majority are only able to do so in continuous flows, restricting the ability to independently control reactions of individual droplets, a prerequisite for programmable digital microfluidics. Here we present a novel method to produce individual picoliter-scale droplets on-demand using surface acoustic waves (SAW). Acoustic forces arising from SAW act on the oil-water interface, creating a droplet whose volume is defined by the applied power, duration of the force and system geometry. Additionally, this method is able to pre-concentrate particles simultaneously with droplet production, meaning that particles and cells, even if in a dilute mixture, can be easily encapsulated. Our method is expected to be applicable to high-throughput screening, bioreactor creation and other microfluidic processes.

Highly Localized Acoustic Streaming and Size-Selective Submicrometer Particle Concentration Using High Frequency Microscale Focused Acoustic Fields

Concentration and separation of particles and biological specimens are fundamental functions of micro/nanofluidic systems. Acoustic streaming is an effective and biocompatible way to create rapid microscale fluid motion and induce particle capture, though the >100 MHz frequencies required to directly generate acoustic body forces on the microscale have traditionally been difficult to generate and localize in a way that is amenable to efficient generation of streaming. Moreover, acoustic, hydrodynamic, and electrical forces as typically applied have difficulty manipulating specimens in the submicrometer regime. In this work, we introduce highly focused traveling surface acoustic waves (SAW) at high frequencies between 193 and 636 MHz for efficient and highly localized production of acoustic streaming vortices on microfluidic length scales. Concentration occurs via a novel mechanism, whereby the combined acoustic radiation and streaming field results in size-selective aggregation in fluid streamlines in the vicinity of a high-amplitude acoustic beam, as opposed to previous acoustic radiation induced particle concentration where objects typically migrate toward minimum pressure locations. Though the acoustic streaming is induced by a traveling wave, we are able to manipulate particles an order of magnitude smaller than possible using the traveling wave force alone. We experimentally and theoretically examine the range of particle sizes that can be captured in fluid streamlines using this technique, with rapid particle concentration demonstrated down to 300 nm diameters. We also demonstrate that locations of trapping and concentration are size-dependent, which is attributed to the combined effects of the acoustic streaming and acoustic forces.

Highly focused high-frequency travelling surface acoustic waves (SAW) for rapid single-particle sorting

High-speed sorting is an essential process in a number of clinical and research applications, where single cells, droplets and particles are segregated based on their properties in a continuous flow. With recent developments in the field of microscale actuation, there is increasing interest in replicating the functions available to conventional fluorescence activated cell sorting (FACS) flow cytometry in integrated on-chip systems, which have substantial advantages in cost and portability. Surface acoustic wave (SAW) devices are ideal for many acoustofluidic applications, and have been used to perform such sorting at rates on the order of kHz. Essential to the accuracy of this sorting, however, is the dimensions of the region over which sorting occurs, where a smaller sorting region can largely avoid inaccurate sorting across a range of sample concentrations. Here we demonstrate the use of flow focusing and a highly focused SAW generated by a high-frequency (386 MHz), 10 μm wavelength set of focused interdigital transducers (FIDTs) on a piezoelectric lithium niobate substrate, yielding an effective sorting region only ~25 μm wide, with sub-millisecond pulses generated at up to kHz rates. Furthermore, because of the use of high frequencies, actuation of particles as small as 2 μm can be realized. Such devices represent a substantial step forward in the evolution of highly localized forces for lab-on-a-chip microfluidic applications.

The particle valve: On-demand particle trapping, filtering, and release from a microfabricated polydimethylsiloxane membrane using surface acoustic waves

We introduce a surface acoustic wave (SAW) based method for acoustically controlled concentration, capture, release, and sorting of particles in a microfluidic system. This method is power efficient by the nature of its design: the vertical direction of a traveling acoustic wave, in which the majority of the energy at the SAW-water interface is directed, is used to concentrate particles behind a microfabricated polydimethylsiloxane membrane extending partially into a channel. Sorting is also demonstrated with this concentration shown to be size-dependent. Low-power, miniature SAW devices, using methods such as the one demonstrated here, are well placed for future integration into point-of-care diagnostic systems.

Detachable Acoustofluidic System for Particle Separation via a Traveling Surface Acoustic Wave

Components in biomedical analysis tools that have direct contact with biological samples, especially biohazardous materials, are ideally discarded after use to prevent cross-contamination. However, a conventional acoustofluidic device is typically a monolithic integration that permanently bonds acoustic transducers with microfluidic channels, increasing processing costs in single-use platforms. In this study, we demonstrate a detachable acoustofluidic system comprised of a disposable channel device and a reusable acoustic transducer for noncontact continuous particle separation via a traveling surface acoustic wave (TSAW). The channel device can be placed onto the SAW transducer with a high alignment tolerance to simplify operation, is made entirely of polydimethylsiloxane (PDMS), and does not require any additional coupling agent. A microstructured pillar is used to couple acoustic waves into the fluid channel for noncontact particle manipulation. We demonstrate the separation of 10 and 15 μm particles at high separation efficiency above 98% in a 49.5 MHz TSAW using the developed detachable acoustofluidic system. Its disposability and ease of assembly should enable broad use of noncontact, disposable particle manipulation techniques in practical biomedical applications related to sample preparation.

Characterization of adhesive properties of red blood cells using surface acoustic wave induced flows for rapid diagnostics

This letter presents a method which employs surface acoustic wave induced acoustic streaming to differentially peel treated red blood cells (RBCs) off a substrate based on their adhesive properties and separate populations of pathological cells from normal ones. We demonstrate the principle of operation by comparing the applied power and time required to overcome the adhesion displayed by healthy, glutaraldehyde-treated or malaria-infected human RBCs. Our experiments indicate that the method can be used to differentiate between various cell populations contained in a 9 μl droplet within 30 s, suggesting potential for rapid diagnostics.

Acoustic tweezers via sub-time-of-flight regime surface acoustic waves.

Researchers use pulsed excitation to generate localized 2D acoustic tweezers for spatially selective microfluidic patterning. Micrometer-scale acoustic waves are highly useful for refined optomechanical and acoustofluidic manipulation, where these fields are spatially localized along the transducer aperture but not along the acoustic propagation direction. In the case of acoustic tweezers, such a conventional acoustic standing wave results in particle and cell patterning across the entire width of a microfluidic channel, preventing selective trapping. We demonstrate the use of nanosecond-scale pulsed surface acoustic waves (SAWs) with a pulse period that is less than the time of flight between opposing transducers to generate localized time-averaged patterning regions while using conventional electrode structures. These nodal positions can be readily and arbitrarily positioned in two dimensions and within the patterning region itself through the imposition of pulse delays, frequency modulation, and phase shifts. This straightforward concept adds new spatial dimensions to which acoustic fields can be localized in SAW applications in a manner analogous to optical tweezers, including spatially selective acoustic tweezers and optical waveguides.

Continuous micro-vortex-based nanoparticle manipulation via focused surface acoustic waves

Despite increasing demand in the manipulation of nanoscale objects for next generation biological and industrial processes, there is a lack of methods for reliable separation, concentration and purification of nanoscale objects. Acoustic methods have proven their utility in contactless manipulation of microscale objects mainly relying on the acoustic radiation effect, though the influence of acoustic streaming has typically prevented manipulation at smaller length scales. In this work, however, we explicitly take advantage of the strong acoustic streaming in the vicinity of a highly focused, high frequency surface acoustic wave (SAW) beam emanating from a series of focused 6 μm substrate wavelength interdigital transducers patterned on a piezoelectric lithium niobate substrate and actuated with a 633 MHz sinusoidal signal. This streaming field serves to focus fluid streamlines such that incoming particles interact with the acoustic field similarly regardless of their initial starting positions, and results in particle displacements that would not be possible with a travelling acoustic wave force alone. This streaming-induced manipulation of nanoscale particles is maximized with the formation of micro-vortices that extend the width of the microfluidic channel even with the imposition of a lateral flow, occurring when the streaming-induced flow velocities are an order of magnitude larger than the lateral one. We make use of this acoustic streaming to demonstrate the continuous and differential focusing of 100 nm, 300 nm and 500 nm particles.

Batch process particle separation using surface acoustic waves (SAW): integration of travelling and standing SAW

Acoustic fields offer a versatile and non-contact method for particle and cell manipulation, where several acoustofluidic systems have been developed for the purpose of sorting. However, in almost all cases, these systems utilize a steady flow to either define the exposure time to the acoustic field or to counteract the acoustic forces. Batch-based systems, within which sorting occurs in a confined volume, are compatible with smaller sample volumes without the need for externally pumped flow, though remain relatively underdeveloped. Here, the effects of utilizing a combination of travelling and standing waves on particles of different sizes are examined. We use a pressure field combining both travelling and standing wave components along with a swept excitation frequency, to collect and isolate particles of different sizes in a static fluid volume. This mechanism is employed to demonstrate size-based deterministic sorting of particles. Specifically, 5.1 μm and 7 μm particles are separated using a frequency range from 60 MHz to 90 MHz, and 5.1 μm particles are separated from 3.1 μm using an excitation sweeping between 70 MHz and 120 MHz.