This article reviews acoustic microfiuidics: the use of acoustic fields, principally ultrasonics, for application in microfiuidics. Although acoustics is a classical field, its promising, and indeed perplexing, capabilities in powerfully manipulating both fluids and particles within those fluids on the microscale to nanoscale has revived interest in it. The bewildering state of the literature and ample jargon from decades of research is reorganized and presented in the context of models derived from first principles. This hopefully will make the area accessible for researchers with experience in materials science, fluid mechanics, or dynamics. The abundance of interesting phenomena arising from nonlinear interactions in ultrasound that easily appear at these small scales is considered, especially in surface acoustic wave devices that are simple to fabricate with planar lithography techniques common in microfluidics, along with the many applications in microfluidics and nanofluidics that appear through the literature.

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Microscale acoustofluidics: Microfluidics driven via acoustics and ultrasonics

Droplet based microfluidics is a rapidly growing interdisciplinary field of research combining soft matter physics, biochemistry and microsystems engineering. Its applications range from fast analytical systems or the synthesis of advanced materials to protein crystallization and biological assays for living cells. Precise control of droplet volumes and reliable manipulation of individual droplets such as coalescence, mixing of their contents, and sorting in combination with fast analysis tools allow us to perform chemical reactions inside the droplets under defined conditions. In this paper, we will review available drop generation and manipulation techniques. The main focus of this review is not to be comprehensive and explain all techniques in great detail but to identify and shed light on similarities and underlying physical principles. Since geometry and wetting properties of the microfluidic channels are crucial factors for droplet generation, we also briefly describe typical device fabrication methods in droplet based microfluidics. Examples of applications and reaction schemes which rely on the discussed manipulation techniques are also presented, such as the fabrication of special materials and biophysical experiments.

https://iopscience.iop.org/article/10.1088/0034-4885/75/1/016601/pdf

Droplet based microfluidics

Accurate and high throughput cell sorting is a critical enabling technology in molecular and cellular biology, biotechnology, and medicine While conventional methods can provide high efficiency sorting in short timescales, advances in microfluidics have enabled the realization of miniaturized devices offering similar capabilities that exploit a variety of physical principles We classify these technologies as either active or passive Active systems generally use external fields (eg, acoustic, electric, magnetic, and optical) to impose forces to displace cells for sorting, whereas passive systems use inertial forces, filters, and adhesion mechanisms to purify cell populations Cell sorting on microchips provides numerous advantages over conventional methods by reducing the size of necessary equipment, eliminating potentially biohazardous aerosols, and simplifying the complex protocols commonly associated with cell sorting Additionally, microchip devices are well suited for parallelization, enabling complete lab-on-a-chip devices for cellular isolation, analysis, and experimental processing In this review, we examine the breadth of microfluidic cell sorting technologies, while focusing on those that offer the greatest potential for translation into clinical and industrial practice and that offer multiple, useful functions We organize these sorting technologies by the type of cell preparation required (ie, fluorescent label-based sorting, bead-based sorting, and label-free sorting) as well as by the physical principles underlying each sorting mechanism

Microfluidic cell sorting: a review of the advances in the separation of cells from debulking to rare cell isolation

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.

https://www.pnas.org/content/pnas/109/28/11105.full.pdf

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

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.

Surface acoustic wave microfluidics

We describe a novel microfluidic cell sorter which operates in continuous flow at high sorting rates. The device is based on a surface acoustic wave cell-sorting scheme and combines many advantages of fluorescence activated cell sorting (FACS) and fluorescence activated droplet sorting (FADS) in microfluidic channels. It is fully integrated on a PDMS device, and allows fast electronic control of cell diversion. We direct cells by acoustic streaming excited by a surface acoustic wave which deflects the fluid independently of the contrast in material properties of deflected objects and the continuous phase; thus the device underlying principle works without additional enhancement of the sorting by prior labelling of the cells with responsive markers such as magnetic or polarizable beads. Single cells are sorted directly from bulk media at rates as fast as several kHz without prior encapsulation into liquid droplet compartments as in traditional FACS. We have successfully directed HaCaT cells (human keratinocytes), fibroblasts from mice and MV3 melanoma cells. The low shear forces of this sorting method ensure that cells survive after sorting.

https://projects.iq.harvard.edu/files/weitzlab/files/2010_labchip_franke.pdf

Surface acoustic wave actuated cell sorting (SAWACS)

Using size and deformability as intrinsic biomarkers, we separate red blood cells (RBCs) from other blood components based on a repulsive hydrodynamic cell-wall-interaction. We exploit this purely viscous lift effect at low Reynolds numbers to induce a lateral migration of soft objects perpendicular to the streamlines of the fluid, which closely follows theoretical prediction by Olla [J. Phys. II 7, 1533, (1997)]. We study the effects of flow rate and fluid viscosity on the separation efficiency and demonstrate the separation of RBCs, blood platelets, and solid microspheres from each other. The method can be used for continuous and label-free cell classification and sorting in on-chip blood analysis.

Separation of blood cells using hydrodynamic lift

The effects of membrane cholesterol and simvastatin on red blood cell deformability and ATP release.

We study the mechanical relaxation behavior of human red blood cells by observing the time evolution of shape change of cells flowing through microchannels with abrupt constrictions. We observe two types of relaxation processes. In the first fast process (τ1 ∼ 200 ms) the initially parachute shaped cells relax into cup-shaped cells (stomatocytes). These cells relax and reorient in a second relaxation process with a response time of τ1/2 ∼ 10 s into the equilibrium discoid shapes. The values for the relaxation times of single red blood cells in the population scatter significantly within the cell population between 0.11 s < τ1 < 0.52 s and 9 s < τ1/2 < 49 s, respectively. However, when plotting τ1/2 against τ1, we find a linear relationship between the two timescales and are able to relate both to the elastic properties of the spectrin cytoskeleton underlying the red cell's plasma membrane. Adenosine Triphosphate (ATP) enhances dissociation of spectrin filaments resulting in a reduced shear modulus. We modify the cytoskeleton connectivity by depletion and repletion of ATP and study the effect on relaxation. Both the linear relationship of timescales as well as the ATP dependence can be understood by theoretical models.

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Hydrodynamic deformation reveals two coupled modes/time scales of red blood cell relaxation

Cryostorage media containing 1% of the non-ionic surfactant Poloxamer 188 provided full recovery of mammalian cells (Gonzalez Hernandez, 2006), but its effects during thawing of cryostored cells and proteolytic subcultivation are still unknown. In this study, the proliferation and viability of pre-confluent Caco-2 monolayers cultivated in media supplemented with the non-ionic surfactant were investigated. The addition of 0.5% Poloxamer 188 increased proliferation of subcultivated cells 1.5 fold and that of thawed cells about twofold. According to microaspiration experiments the non-ionic surfactant increased the tension of the cell membrane most notably at concentrations = 0.5% owing to adsorption and incorporation into the phospholipid bilayer. Thus, the performance of the cells appears to be improved. Since vitality of cells is a prerequisite for reproducibility and reliability of cell models for absorption studies at early stages of drug development, use of Poloxamer 188 supplemented cultivation media may help to refine cell culturing to further reduce animal trials in preclinical investigations.

Susanne Braunmüller

Papers

Surface acoustic wave actuated cell sorting (SAWACS)

Separation of blood cells using hydrodynamic lift

The effects of membrane cholesterol and simvastatin on red blood cell deformability and ATP release.

Hydrodynamic deformation reveals two coupled modes/time scales of red blood cell relaxation

Poloxamer 188 supplemented culture medium increases the vitality of Caco-2 cells after subcultivation and freeze/thaw cycles.