Quantifying cell adhesion through forces generated by acoustic streaming.

The strength of cell adhesion is important in understanding the cell's health and in culturing them. Quantitative measurement of cell adhesion strength is a significant challenge in bioengineering research. For this, the present study describes a system that can measure cell adhesion strength using acoustic streaming induced by Lamb waves. Cells are cultured on an ultrasound transducer using a range of preculture and incubation times with phosphate-buffered saline (PBS) just before the measurement. Acoustic streaming is then induced using several Lamb wave intensities, exposing the cells to shear flows and eventually detaching them. By relying upon a median detachment rate of 50 %, the corresponding detachment force, or force of cell adhesion, was determined to be on the order of several nN, consistent with previous reports. The stronger the induced shear flow, the more cells were detached. Further, we employed a preculture time of 8 to 24 h and a PBS incubation time of 0 to 60 min, producing cell adhesion forces that varied from 1.2 to 13 nN. Hence, the developed system can quantify cell adhesion strength over a wide range, possibly offering a fundamental tool for cell-based bioengineering.

Acousto-Photolithography for Programmable Shape Deformation of Composite Hydrogel Sheets.

Stimuli-responsive hydrogels with programmable shapes produced by defined patterns of particles are of great interest for the fabrication of small-scale soft actuators and robots. Patterning the particles in the hydrogels during fabrication generally requires external magnetic or electric fields, thus limiting the material choice for the particles. Acoustically driven particle manipulation, however, solely depends on the acoustic impedance difference between the particles and the surrounding fluid, making it a more versatile method to spatially control particles. Here, an approach is reported by combining direct acoustic force to align photothermal particles and photolithography to spatially immobilize these alignments within a temperature-responsive poly(N-isopropylacrylamide) hydrogel to trigger shape deformation under temperature change and light exposure. The spatial distribution of particles can be tuned by the power and frequency of the acoustic waves. Specifically, changing the spacing between the particle patterns and position alters the bending curvature and direction of this composite hydrogel sheet, respectively. Moreover, the orientation (i.e., relative angle) of the particle alignments with respect to the long axis of laser-cut hydrogel strips governs the bending behaviors and the subsequent shape deformation by external stimuli. This acousto-photolithography provides a means of spatiotemporal programming of the internal heterogeneity of composite polymeric systems.

Well-free agglomeration and on-demand three-dimensional cell cluster formation using guided surface acoustic waves through a couplant layer.

Three-dimensional cell agglomerates are broadly useful in tissue engineering and drug testing. We report a well-free method to form large (1.4-mm) multicellular clusters using 100-MHz surface acoustic waves (SAW) without direct contact with the media or cells. A fluid couplant is used to transform the SAW into acoustic streaming in the cell-laden media held in a petri dish. The couplant transmits longitudinal sound waves, forming a Lamb wave in the petri dish that, in turn, produces longitudinal sound in the media. Due to recirculation, human embryonic kidney (HEK293) cells in the dish are carried to the center of the coupling location, forming a cluster in less than 10 min. A few minutes later, these clusters may then be translated and merged to form large agglomerations, and even repeatedly folded to produce a roughly spherical shape of over 1.4 mm in diameter for incubation-without damaging the existing intercellular bonds. Calcium ion signaling through these clusters and confocal images of multiprotein junctional complexes suggest a continuous tissue construct: intercellular communication. They may be formed at will, and the method is feasibly useful for formation of numerous agglomerates in a single petri dish.

Focused surface acoustic wave locally removes cells from culture surface.

Regenerative medicine and drug development require large numbers of high-quality cells, usually delivered from in vitro culturing. During culturing, the appearance of unwanted cells and an inability to remove them without damaging or losing most if not all the surrounding cells in the culture reduce the overall quality of the cultured cells. This is a key problem in cell culturing, as is the inability to sample cells from a culture as desired to verify the quality of the culture. Here, we report a method to locally remove cells from an adherent cell culture using a 100.4 MHz focused surface acoustic wave (SAW) device. After exposing a plated C2C12 mouse myoblast cell culture to phosphate buffered solution (PBS), ultrasound from the SAW device transmitted into the cell culture via a coupling water droplet serves to detach a small grouping of cells. The cells are removed from an area 6 × 10-3 mm2, equivalent to about 12 cells, using a SAW device-Petri dish water gap of 1.5 mm, a PBS immersion time of 300 s, and an input voltage of 75 V to the SAW device. Cells were released as desired 90% of the time, releasing the cells from the target area nine times out of ten runs. In the one trial in ten that fails, the cells partially release and remain attached due to inter-cellular binding. By making it possible to target and remove small groups of cells as desired, the quality of cell culturing may be significantly improved. The small group of cells may be considered a colony of iPS cells. This targeted cell removal method may facilitate sustainable, contamination-free, and automated refinement of cultured cells.

Practical microcircuits for handheld acoustofluidics.

Acoustofluidics has promised to enable lab-on-a-chip and point-of-care devices in ways difficult to achieve using other methods. Piezoelectric ultrasonic transducers-as small as the chips they actuate-provide rapid fluid and suspended object transport. Acoustofluidic lab-on-chip devices offer a vast range of benefits in early disease identification and noninvasive drug delivery. However, their potential has long been undermined by the need for benchtop or rack-mount electronics. The piezoelectric ultrasonic transducers within require these equipment and thus acoustofluidic device implementation in a bedside setting has been limited. Here we detail a general process to enable the reader to produce battery or mains-powered microcircuits ideal for driving 1-300 MHz acoustic devices. We include the general design strategy for the circuit, the blocks that collectively define it, and suitable, specific choices for components to produce these blocks. We furthermore illustrate how to incorporate automated resonance finding and tracking, sensing and feedback, and built-in adjustability to accommodate devices' vastly different operating frequencies and powers in a single driver, including examples of fluid and particle manipulation typical of the needs in our discipline. With this in hand, the many groups active in lab-on-a-chip acoustofluidics can now finally deliver on the promise of handheld, point-of-care technologies.

Fabrication of Surface Acoustic Wave Devices on Lithium Niobate.

Manipulation of fluids and particles by acoustic actuation at small scale is aiding the rapid growth of lab-on-a-chip applications. Megahertz-order surface acoustic wave (SAW) devices generate enormous accelerations on their surface, up to 108 m/s2, in turn responsible for many of the observed effects that have come to define acoustofluidics: acoustic streaming and acoustic radiation forces. These effects have been used for particle, cell, and fluid handling at the microscale-and even at the nanoscale. In this paper we explicitly demonstrate two major fabrication methods of SAW devices on lithium niobate: the details of lift-off and wet etching techniques are described step-by-step. Representative results for the electrode pattern deposited on the substrate as well as the performance of SAW generated on the surface are displayed in detail. Fabrication tricks and troubleshooting are covered as well. This procedure offers a practical protocol for high frequency SAW device fabrication and integration for future microfluidics applications.

Optimized, Omnidirectional Surface Acoustic Wave Source: 152° Y-Rotated Cut of Lithium Niobate for Acoustofluidics.

Here, we propose an optimized Y -rotated cut of lithium niobate (LN) for multidirectional surface acoustic wave (SAW) propagation, simultaneously minimizing the anisotropic effects while maximizing the electromechanical properties of this cut of LN. The goal is to offer a piezoelectric material suitable for acoustofluidics applications that require greater flexibility in wave generation and propagation than the currently ubiquitous 128° Y -rotated X -propagating cut. The 128YX LN cut is known to most effectively generate Rayleigh SAW along the x -direction alone. Any SAW veering from this propagation direction is affected by beam steering and changes in resonance frequency and electromechanical coupling coefficients, consequently limiting the use of LN in various acoustofluidics applications, where more diverse configurations would be beneficial. The L2 -norm of these properties was evaluated under rotational transformation to produce a physical model with closed governing equations for 40-MHz surface wave propagation on the surface of a piezoelectric material. This was then utilized to obtain the surface wave velocity and coupling coefficient of the specific Y -cut LN with respect to the propagating direction. Next, the averaged coupling coefficients of various Y -cuts of LN in all propagating directions were calculated and integrated to simultaneously minimize anisotropy and maximize the electromechanical properties of the LN substrate. A 152° Y -rotated cut was found to be the optimal choice under these constraints, enabling multidirectional SAW propagation with greater coupling and lower variation in wave performance for SAW generated across the surface in any direction. Compared with the 128YX LN cut, this cut provides a 66.5% improvement in the in-plane isotropy and a 37.0% improvement in the average electromechanical coupling for in-plane SAW propagation. Experimental devices operating at the frequency of 40 MHz were designed, fabricated, and tested on the surface of this 500- [Formula: see text]-thick specific cut of LN and served to verify the supporting analysis and the superior isotropic properties of the 152° Y -rotated cut in generating SAW.

Micro/nano acoustofluidics: materials, phenomena, design, devices, and applications.

Acoustic actuation of fluids at small scales may finally enable a comprehensive lab-on-a-chip revolution in microfluidics, overcoming long-standing difficulties in fluid and particle manipulation on-chip. In this comprehensive review, we examine the fundamentals of piezoelectricity, piezoelectric materials, and transducers; revisit the basics of acoustofluidics; and give the reader a detailed look at recent technological advances and current scientific discussions in the discipline. Recent achievements are placed in the context of classic reports for the actuation of fluid and particles via acoustic waves, both within sessile drops and closed channels. Other aspects of micro/nano acoustofluidics are examined: atomization, translation, mixing, jetting, and particle manipulation in the context of sessile drops and fluid mixing and pumping, particle manipulation, and formation of droplets in the context of closed channels, plus the most recent results at the nanoscale. These achievements will enable applications across the disciplines of chemistry, biology, medicine, energy, manufacturing, and we suspect a number of others yet unimagined. Basic design concepts and illustrative applications are highlighted in each section, with an emphasis on lab-on-a-chip applications.

Magnetically separable Prussian blue analogue Mn3[Co(CN)6]2·nH2O porous nanocubes as excellent absorbents for heavy metal ions.

The application of Prussian blue analogue (PBA) Mn(3)[Co(CN)(6)](2)·nH(2)O porous nanocubes as absorbents for heavy metal ions has been demonstrated. The result indicates that Mn(3)[Co(CN)(6)](2)·nH(2)O porous nanocubes with average diameter of 240 nm possess excellent adsorption efficiency for Pb(2+) ions (94.21% at initial Pb(2+) concentration of 10 mg L(-1)). Moreover, Mn(3)[Co(CN)(6)](2)·nH(2)O porous nanocubes can also show high adsorption efficiency on heavy metal ions even in a strong acidic solution due to its chemical stability. Notably, an external magnet could be used to accelerate the separation of Mn(3)[Co(CN)(6)](2)·nH(2)O from the treated solution. It is suggested that the high adsorption efficiency may derive from the large surface area, M(3)(II)[M(III)(CN)(6)](2)·nH(2)O porous framework structure and affinity between polarizable π-electron clouds of the cyanide bridges and heavy metals ions.

Prussian Blue Analogue Mn3[Co(CN)6]2·nH2O porous nanocubes: large-scale synthesis and their CO2 storage properties.

Prussian Blue Analogue (PBA) Mn(3)[Co(CN)(6)](2)·nH(2)O porous nanocubes were successfully synthesized in high yield at room temperature in the presence of poly(vinylpyrrolidone) (PVP) and characterized by X-ray powder diffraction (XRD), field emission scanning electron microscopy (FE-SEM), and transmission electron microscopy (TEM). The effects of synthetic parameters such as surfactant, the ratio of different solvents on the morphology and size of the particles were investigated. The experimental results showed that poly(vinylpyrrolidone) (PVP) and solvent ethanol play critical roles in the formation of uniform porous nanocubes. N(2) adsorption properties indicated that the Mn(3)[Co(CN)(6)](2) porous nanocubes with an average diameter of 240 nm possessed a high surface area of 675 m(2) g(-1) with total volume of 0.354 cm(3) g(-1). Moreover, the porous nanocubes showed high CO(2) adsorption at room temperature and 1 bar of pressure. To our knowledge, this is the first report on the synthesis of Mn(3)[Co(CN)(6)](2) nanomaterials and their CO(2) adsorption applications at the nanoscale.