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ACOUSTOFLUIDICS: TECHNIQUES FOR MICROFLUIDIC PARTICLE MANIPULATION

This review explores the interdisciplinary field of acoustofluidics, which merges acoustics with microfluidics to manipulate fluids and particles at the microscale using sound waves. It examines fundamental concepts, including acoustic wave types such as Surface Acoustic Waves (SAW) and Bulk Acoustic Waves (BAW). Applications in microfluidic devices like particle sorting, mixing, biosensing, and medium switching are emphasized. Additionally, the review addresses the advantages, challenges, and future directions of these technologies, emphasizing their role in advancing lab-on-a-chip innovations and enhancing microfluidic device capabilities. Notable recent advances include single-cell analysis, point-of-care diagnostics, and tissue engineering applications.

What is Acoustofluidics?

Acoustofluidics is an interdisciplinary field that combines acoustics and microfluidics to manipulate fluids and particles at the microscale using sound waves. Emerging from advancements in microfabrication, it allows for precise, contactless control over processes such as acoustic particle sorting and separation based on properties like shape, flexibility, and activity. This makes it invaluable in medical, biological, and chemical analyses. Acoustic waves, owing to their efficiency and biocompatibility, are particularly effective for sorting particles of all sizes. Recent advancements have also integrated dynamic field manipulation and multi-resonator designs to expand its capabilities.

Acoustofluidics principles adapted from (Mengxi Wu et al., 2019)

 

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Physics Behind Acoustofluidics

Acoustofluidics involves the manipulation of particles and fluids using acoustic waves, which are mechanical vibrations traveling through air, liquid, or solid media. These waves generate forces that control particle movement, separation, and concentration at the microscale. The tunability and non-invasive nature of these forces make acoustofluidics particularly useful in biological and chemical applications. Key factors influencing these interactions include wave frequency, amplitude, particle properties (size, density, compressibility), and the propagation medium. Understanding these principles is crucial for optimizing device performance in complex systems. Acoustic forces include:

  • Acoustic Radiation Force: Arising from scattering and absorption, this force transfers linear momentum from the acoustic field to the object. Gor’kov’s potential is often used to predict these forces based on particle size, density, and compressibility.
  • Acoustic Streaming: A nonlinear phenomenon resulting from acoustic wave attenuation in a viscous medium. This streaming enhances fluid mixing and particle motion.
  • Buoyancy and Drag Forces: These forces act alongside acoustic forces in fluid flow, with drag forces influenced by streaming velocities.
  • Other Field-Induced Forces: Examples include dielectrophoretic forces and optical forces, which may complement acoustic manipulation in hybrid systems.

Recent computational approaches, such as finite element methods (e.g., COMSOL), have improved the modelling and understanding of these forces, aiding in device optimization.

Types of Waves Used for Acoustofluidics

Acoustofluidics employs two primary wave types:

  1. Surface Acoustic Waves (SAW): Propagate along a substrate’s surface and operate in the megahertz frequency range. They include:
    • Traveling SAW (TSAW): Waves travel in a single direction.
    • Standing SAW (SSAW): Interfering waves form nodes and antinodes. Advanced implementations use holographic SAW transducers for dynamic pressure distributions.
  2. Bulk Acoustic Waves (BAW): Travel through the bulk of a medium and are generated by piezoelectric transducers. These waves are effective for larger fluid volumes and operate within the medium itself. Multi-element transducer arrays now allow dynamic adjustments of acoustic fields for specific applications.
Schematic summarizing different acoustic waves adapted from Surface-acoustic-wave (SAW) and bulk-acoustic-wave (BAW)
Schematic summarizing different acoustic waves adapted from Surface-acoustic-wave (SAW) and bulk-acoustic-wave (BAW) Filters will be disruptor in the military RF and microwave industries, https://idstch.com/, Rajesh Uppal, 2021

Acoustofluidic Setups

Acoustofluidic systems are highly specialized assemblies that integrate cutting-edge components to precisely manipulate fluids and particles at the microscale. These systems combine mechanical precision with optical monitoring, making them versatile tools for research and industrial applications ranging from biological analysis to chemical processing. 

  • A fluidic source (e.g., syringe pump, flow controller).
  • A microfluidic chip with a piezoelectric acoustic transducer.
  • A signal generator for acoustic wave production.
  • An optical microscope for visualization.

Modern setups integrate laser Doppler vibrometers and hydrophones for precise field measurements. Additionally, metasurfaces and holographic elements enable tailored acoustic field patterns for complex applications.

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Applications for Acoustofluidics

Acoustofluidics applications

Cell and Particle Sorting

Cell sorting using acoustofluidics provides a contact-free and biocompatible method for separating cells based on their size, density, and mechanical properties. This approach is particularly valuable for applications where maintaining cell viability and function is critical, such as isolating rare cell populations (e.g., circulating tumor cells or stem cells) for diagnostic and therapeutic purposes. The ability to sort cells efficiently and gently without the use of harsh chemicals or mechanical stress ensures that the sorted cells retain their natural characteristics, making them suitable for downstream applications in research, medicine, and biotechnology. There are several ways for cell sorting using acoustofluidics:

Free-Flow Acoustophoresis

Particles move to pressure nodes based on size, with larger particles experiencing stronger acoustic radiation forces. Tilted-angle standing SAWs improve sorting efficiency, and advanced designs now handle highly concentrated solutions effectively.

Seeding Beads-Assisted Separation

Preloaded micron-sized particles trap targets via secondary radiation forces, allowing separation based on density and compressibility. Acoustic contrast factor-based binary separations are widely applied in medical diagnostics and lipid emboli removal. (Persson et al., 2008)

Manipulating Frequencies of Acoustic Waves

By tuning wave frequencies, particles align at specific pressure nodes. Multi-resonator designs allow simultaneous sorting of multiple populations, improving throughput for complex biological samples.

Separating blood cells from plasma by acoustic waves for the applications
Separating blood cells from plasma by acoustic waves for the applications of blood wash or plasmapheresis (Mengxi Wu, Microsystems & Nanoengineering 2019) https://www.nature.com/articles/s41378-019-0064-3

3D Cell Culture

Acoustic waves play a critical role in advancing 3D cell culture techniques, particularly for tissue engineering and organoid assembly. By leveraging acoustic forces, researchers can aggregate and pattern cells with precision, facilitating the creation of complex tissue structures. Acoustic streaming further enhances these processes by enabling efficient scaffold patterning and the manipulation of organoids in a contact-free manner. This approach ensures minimal disruption to cell integrity while promoting the assembly of functional biological tissues for research and therapeutic applications.

Single Cell Analysis

Acoustic tweezers are advanced tools that use highly focused acoustic waves to manipulate individual particles or cells with precision. By generating localized acoustic radiation forces, these tweezers can trap and move cells without direct contact, ensuring minimal physical or chemical interference. Acoustic tweezers provide a powerful tool for the precise manipulation of individual cells, enabling detailed studies of cell-cell interactions and signaling. Recent advancements, such as acoustical vortex tweezers, offer enhanced selectivity and minimal biological impact, preserving cell viability during manipulation. Additionally, acoustic force spectroscopy has emerged as a valuable technique for probing the mechanical properties of cells, including measurements of T-cell adhesion strength and red blood cell deformability, providing insights into cellular biomechanics and pathophysiology.

Acoustic tweezers model from Duke’s Acoustofluidics Lab, The Inset indicates a single particle within a “3D trapping node,” which is independently manipulated along the x, y, or z axes.
Acoustic tweezers model from Duke’s Acoustofluidics Lab, The Inset indicates a single particle within a “3D trapping node,” which is independently manipulated along the x, y, or z axes.

Automation Methods

Acoustofluidics enhances liquid handling for applications like point-of-care diagnostics, reducing contamination risks and improving workflow efficiency. An example is liquid biopsy, where recent methods differentiate nanometer- to micrometer-sized particles, such as liposomes, exosomes, and circulating tumor cells.

Fluid Mixing

Fluid mixing is a critical process in microfluidic systems, and acoustofluidics provides an innovative approach to enhance mixing efficiency. Acoustic waves, when applied to a fluid, generate acoustic streaming patterns that create localized vortices and enhance fluid movement. These streaming patterns overcome the challenges of laminar flow typically observed in microfluidic channels, where mixing is limited by diffusion. By employing sharp-edged structures or asymmetric geometries in the microchannel design, the acoustic streaming effect is amplified, leading to more efficient and rapid mixing of reactants. This capability is particularly valuable in chemical reactions, biochemical assays, and sample preparation, where uniform mixing is essential for accuracy and reproducibility. Additionally, acoustofluidic mixing methods are contact-free and do not require mechanical agitation, preserving the integrity of sensitive biological samples or delicate chemical reactions. The ability to precisely control mixing parameters by adjusting wave frequency and amplitude further enhances the versatility of this technique, making it indispensable for applications in diagnostics, drug delivery, and chemical synthesis.

Buffer Switching

Using acoustic waves is also a good way to change particles from a solution to another. The flows in microfluidic channels are laminar. It is then possible to have two laminar streams flowing parallel to each other in a microfluidic channel. By applying an acoustic wave, it is possible to move particles from one stream to the other. The medium/buffer switch is equivalent to a centrifugal wash. Different designs to complete the medium/buffer switch have been reported: a layered resonator, a microfluidic channel with two inlets and two outlets, a transversal resonator and a device with several inlets and outlets located on the side of the channel. (Hawkes et al., 2004)

The use of acoustophoresis reduces the analysis time and the risk of errors as several centrifugation steps are not needed any more. This technique is also of great interest in order to pull out rare species present in complex solutions. 

Advantages and Challenges of Acoustofluidics

Acoustofluidics pros and cons

Acoustofluidics offers transformative benefits that make it an essential technology in modern microfluidics, enabling precise and efficient manipulation of fluids and particles across various scientific and industrial domains:

  • Non-Invasive Operation: Acoustofluidics offers unparalleled precision in manipulating fluids and particles without direct contact, significantly reducing contamination risks and ensuring sample purity.
  • Biocompatibility: The gentle nature of acoustic forces ensures minimal impact on biological samples, preserving cell viability and functionality even during complex manipulations.
  • Versatility: Capable of handling a vast range of particle sizes and fluid types, acoustofluidic devices demonstrate unmatched adaptability across diverse applications, from medical diagnostics to industrial processing.

Acoustofluidics faces several challenges that require attention for widespread adoption and optimization:

  • Complex Device Design and Data Reproducibility: The design of devices is inherently complex, demanding precise fabrication and meticulous tuning to achieve desired performance while balancing cost and scalability. Additionally, the absence of standardized protocols undermines the reliability and comparability of results, posing a significant hurdle for collaborative research and industrial applications.
  • Energy Efficiency and Heat Management: Managing power consumption and heat dissipation is critical to maintaining device functionality and ensuring the integrity of biological samples during operation.
  • Standardization and Technical Expertise Requirements: The lack of standardized methods to evaluate the biological effects of acoustic waves leads to inconsistencies in experimental results across laboratories, complicating reproducibility. Furthermore, operating these systems necessitates high technical expertise, which can limit accessibility and broader adoption.

Conclusion

Acoustofluidics stands at the intersection of acoustics and microfluidics, offering revolutionary methods for microscale particle and fluid manipulation. Applications like particle sorting, biosensing, and fluid mixing highlight its importance in scientific research. Addressing challenges such as energy efficiency and standardization will drive further innovation, unlocking new capabilities for microfluidic devices. The integration of advanced tools, such as dynamic field manipulation and metasurfaces, is paving the way for transformative progress in biomedical diagnostics and tissue engineering.

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References
  • M. Wu, et al., Principles of Acoustofluidics, Microsystems & Nanoengineering, Nature, 2019. Available: https://www.nature.com/articles/s41378-019-0064-3
  • R. Uppal, Surface-acoustic-wave (SAW) and bulk-acoustic-wave (BAW) Filters as Disruptors in Military RF and Microwave Industries, IDST, 2021. Available: https://idstch.com/technology/electronics/surface-acoustic-wave-saw-bulk-acoustic-wave-baw-filters-will-disruptor-military-rf-microwave-industries/
  • Persson, et al., Applications of Bead Affinity in Acoustic Medium Switching, 2008.
  • Hawkes, et al., Layered Resonators for Medium Switching in Microfluidic Devices, 2004.
  • Petersson, et al., Transversal Resonator Design for Particle Focusing in Microfluidics, 2005.
  • Lenshof, et al., Acoustic Particle Sorting Based on Size and Density, 2012.
  • Grenvall, et al., Separation of Lipid Particles in Milk by Acoustic Contrast Factors, 2009.
  • Gupta, et al., Binary Acoustic Contrast Separation in Microfluidic Systems, 1995.
  • Augustsson, et al., Techniques for Measuring Compressibility and Density of Particles in Acoustofluidic Systems, 2011.
  • Adams and Soh, Acoustic “Band-Pass” Particle Size Filtering, 2010.
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