Acoustofluidics-Assisted Multifunctional Paper-Based Analytical Devices.

Microfluidic paper-based analytical devices (µPADs) feature an economic and sensitive nature, while acoustofluidics displays contactless and versatile virtue, and both of them gained tremendous interest in the past decades. Integrating µPADs with acoustofluidic techniques provides great potential to overcome the inherent shortcomings and make appealing achievements. Here, we present acoustofluidics-assisted multifunctional paper-based analytical devices that leverage bulk acoustic waves to realize multiple applications on paper substrates, including uniform colorimetric detection, microparticle/cell enrichment, fluorescence amplification, homogeneous mixing, and nanomaterial synthesis. The glucose detection in the range of 5-15 mM was conducted to perform uniform colorimetric detection. Various types (brass powder, copper powder, diamond powder, and yeast cells) and sizes (5-200 µm) of solid particles and biological cells can be enriched on paper in a few seconds or minutes; thus, fluorescence amplification by 3 times was realized with the enrichment. The high-throughput and homogeneous mixing of two fluids can be achieved, and based on the mixing, nanomaterials (ZnO nanosheets) were synthesized on paper. We analyzed the underlying mechanisms of these applications in the devices, which are attributed to Faraday waves and Chladni patterns. With their simple fabrication and prominent effectiveness, the devices open up new possibilities for paper-based microfluidic devices.

Rational Design of Liquid-Liquid Microdispersion Droplet Microreactors for the Controllable Synthesis of Highly Uniform and Monodispersed Dextran Microspheres.

Hydrogel microspheres are biocompatible materials widely used in biological and medical fields. Emulsification and stirring are the commonly used methods to prepare hydrogels. However, the size distribution is considerably wide, the monodispersity and the mechanical intensity are poor, and the stable operation conditions are comparatively narrow to meet some sophisticated applications. In this paper, a T-shaped stepwise microchannel combined with a simple side microchannel structure is developed to explore the liquid-liquid dispersion mechanism, interfacial evolution behavior, satellite droplet formation mechanism and separation, and the eventual successful synthesis of dextran hydrogel microspheres. The effect of the operation parameters on droplet and microsphere size is comprehensively studied. The flow pattern and the stable operation condition range are given, and mathematical prediction models are developed under three different flow regimes for droplet size prediction. Based on the stable operating conditions, a microdroplet-based method combined with UV light curing is developed to synthesize the dextran hydrogel microsphere. The highly uniform and monodispersed dextran microspheres with good mechanical intensity are synthesized in the developed microfluidic platform. The size of the microsphere could be tuned from 50 to 300 µm with a capillary number in the range of 0.006-0.742. This work not only provides a facile method for functional polymeric microsphere preparation but also offers important design guidelines for the development of a robust microreactor.

Capillary-based, multifunctional manipulation of particles and fluids via focused surface acoustic waves.

Surface acoustic wave (SAW)-enabled acoustofluidic technologies have recently atttracted increasing attention for applications in biology, chemistry, biophysics, and medicine. Most SAW acoustofluidic devices generate acoustic energy which is then transmitted into custom microfabricated polymer-based channels. There are limited studies on delivering this acoustic energy into convenient commercially-available glass tubes for manipulating particles and fluids. Herein, we have constructed a capillary-based SAW acoustofluidic device for multifunctional fluidic and particle manipulation. This device integrates a converging interdigitated transducer to generate focused SAWs on a piezoelectric chip, as well as a glass capillary that transports particles and fluids. To understand the actuation mechanisms underlying this device, we performed finite element simulations by considering piezoelectric, solid mechanic, and pressure acoustic physics. This experimental study shows that the capillary-based SAW acoustofluidic device can perform multiple functions including enriching particles, patterning particles, transporting particles and fluids, as well as generating droplets with controlled sizes. Given the usefulness of these functions, we expect that this acoustofluidic device can be useful in applications such as pharmaceutical manufacturing, biofabrication, and bioanalysis.

Fabrication of monodisperse magnetic nanorods for improving hyperthermia efficacy.

BACKGROUND: Hyperthermia is one of the promising cancer treatment strategies enabled by local heating with the use of tumor-targeting magnetic nanoparticles (MNP) under a non-invasive magnetic field. However, one of the remaining challenges is how to achieve therapeutic levels of heat (without causing damages to regular tissues) in tumors that cannot be effectively treated with anti-tumor drug delivery. RESULTS: In this work, we report a facile method to fabricate magnetic nanorods for hyperthermia by one-step wet chemistry synthesis using 3-Aminopropyltrimethoxysilane (APTMS) as the shape-controlling agent and ferric and ferrous ions as precursors. By adjusting the concentration of APTMS, hydrothermal reaction time, ratios of ferric to ferrous ions, magnetic nanorods with aspect ratios ranging from 4.4 to 7.6 have been produced. At the clinically recommended field strength of 300 Oe (or less) and the frequency of 184 kHz, the specific absorption rate (SAR) of these nanorods is approximately 50 % higher than that of commercial Bionized NanoFerrite particles. CONCLUSIONS: This increase in SAR, especially at low field strengths, is crucial for treating deep tumors, such as pancreatic and rectal cancers, by avoiding the generation of harmful eddy current heating in normal tissues.

Acoustofluidics-Assisted Fluorescence-SERS Bimodal Biosensors.

Acoustofluidics, the fusion of acoustics and microfluidic techniques, has recently seen increased research attention across multiple disciplines due in part to its capabilities in contactless, biocompatible, and precise manipulation of micro-/nano-objects. Herein, a bimodal signal amplification platform which relies on acoustofluidics-induced enrichment of nanoparticles is introduced. The dual-function biosensor can perform sensitive immunofluorescent or surface-enhanced Raman spectroscopy (SERS) detection. The platform functions by using surface acoustic waves to concentrate nanoparticles at either the center or perimeter of a glass capillary; the concentration location is adjusted simply by varying the input frequency. The immunofluorescence assay is achieved by concentrating fluorescent analytes and functionalized nanoparticles at the center of the microchannel, thereby improving the visibility of the fluorescent output. By modifying the inner wall of the glass capillary with plasmonic Ag nanoparticle-deposited ZnO nanorod arrays and focusing analytes toward the perimeter of the microchannel, SERS sensing using the same device setup is achieved. Nanosized exosomes are used as a proof-of-concept to validate the performance of the acoustofluidic bimodal biosensor. With its sample-enrichment functionality, bimodal sensing, short processing time, and minute sample consumption, the acoustofluidic chip holds great potential for the development of lab-on-a-chip based analysis systems in many real-world applications.

Flexible Piezoelectric Nanogenerators Using Metal-doped ZnO-PVDF Films.

Piezoelectric nanomaterial-polymer composites represent a unique paradigm for making flexible energy harvesting and sensing devices with enhanced devices' performance. In this work, we studied various metal doped ZnO nanostructures, fabricated and characterized ZnO nanoparticle-PVDF composite thin film, and demonstrated both enhanced energy generation and motion sensing capabilities. Specifically, a series of flexible piezoelectric nanogenerators (PENGs) were designed based on these piezoelectric composite thin films. The voltage output from cobalt (Co), sodium (Na), silver (Ag), and lithium (Li) doped ZnO-PVDF composite as well as pure ZnO-PVDF samples were individually studied and compared. Under the same experimental conditions, the Li-ZnO based device produces the largest peak-to-peak voltage (3.43 Vpp) which is about 9 times of that of the pure ZnO based device, where Co-ZnO, Na-ZnO and Ag-ZnO are 1.2, 4.9 and 5.4 times, respectively. In addition, the effect of doping ratio of Li-ZnO is studied, and we found that 5% is the best doping ratio in terms of output voltage. Finally, we demonstrated that the energy harvested by the device from finger tapping at ~2 Hz can charge a capacitor with a large output power density of 0.45 W/cm3 and light up an ultraviolet (UV) light-emitting diode (LED). We also showed the device as a flexible wearable motion sensor, where different hand gestures were detected by the device with distinctive output voltage amplitudes and patterns.

Microfluidic continuous flow synthesis of functional hollow spherical silica with hierarchical sponge-like large porous shell.

Microfluidics brings unique opportunities for engineering micro-/nanomaterials with well-controlled physicochemical properties. Herein, using a miniaturized multi-run spiral-shaped microreactor, we develop a flow synthesis strategy to continuously produce hollow spherical silica (HSS) with hierarchical sponge-like pore sizes ranging from several nanometers to over one hundred nanometers. The formation of HSS is realized by mixing two reactant flows, one containing cetyltrimethylammonium bromide (CTAB) and diluted ammonia and the other 1,3,5-trimethylbenzene (TMB) and diluted tetraethyl orthosilicate (TEOS), at a flow rate as high as 5 mL/min. The effect of the reactant concentration and the flow rate on the structural change of the resultant materials is examined. Functional small-sized nanoparticles (magnetic nanoparticle, quantum dot, and silver nanoparticle) can be separately assembled into HSS and high molecular weight protein (bovine serum albumin) can be successfully loaded into HSS and delivered into cancer cells afterward, making them promising in the fields of separation and purification, bioimaging, catalysis, and theranostics.

Microfluidics-enabled rational design of ZnO micro-/nanoparticles with enhanced photocatalysis, cytotoxicity, and piezoelectric properties.

Microfluidics-based reactors enables the controllable synthesis of micro-/nanostructures for a broad spectrum of applications from materials science, bioengineering to medicine. In this study, we first develop a facile and straightforward flow synthesis strategy to control zinc oxide (ZnO) of different shapes (sphere, ellipsoid, short rod, long rod, cube, urchin, and platelet) on a few seconds time scale, based on the 1.5-run spiral-shaped microfluidic reactor with a relative short microchannel length of ca. 92 mm. The formation of ZnO is realized simply by mixing reactants through two inlet flows, one containing zinc nitrate and the other sodium hydroxide. The structures of ZnO are tuned by choosing appropriate flow rates and reactant concentrations of two inlet fluids. The formation mechanism behind microfluidics is proposed. The photocatalysis, cytotoxicity, and piezoelectric capabilities of as-synthesized ZnO from microreactors are further examined, and the structure-dependent efficacy is observed, where higher surface area ZnO structures generally behave better performance. These results bring new insights not only in the rational design of functional micro-/nanoparticles from microfluidics, but also for deeper understanding of the structure-efficacy relationship when translating micro-/nanomaterials into practical applications.

Biomimetic hierarchical walnut kernel-like and erythrocyte-like mesoporous silica nanomaterials: controllable synthesis and versatile applications.

We developed a facile and controllable strategy to fabricate biomimic walnut kernel-like mesoporous silica nanomaterial (WMSN) and erythrocyte-like mesoporous silica nanomaterial (EMSN). The former possesses unique multi-shell hollow structure and surface wrinkles while the latter has special multi-stack structure and bowl-shaped depression. These hierarchical materials with distinct structures can be finely tuned by changing the molar ratios of two surfactants, cetyltrimethylammonium bromide and 11-mercaptoundecanoic acid. The mechanism of structural formation through intermolecular interactions was revealed and validated experimentally. The promising potential applications of WMSN and EMSN in adsorption, cellular imaging, drug delivery, and cancer theranostics were further identified.

Acoustophoresis-driven particle focusing and separation with standard/inverse Chladni patterns.

Manipulating objects with acoustics has been developed for hundreds of years since Chladni patterns in gaseous environments were exhibited. In recent decades, acoustic manipulation in microfluidics, known as acoustofluidics, has rapidly thrived and many sophisticated technologies were born. However, the basic background motion of particles under acoustic excitation is usually neglected and the classical Chladni patterns haven't been reproduced in an aqueous environment. In this study, we investigated the basic mechanism and the motion of suspended particles and sinking particles in a plain microchamber under low-frequency excitation (3-5 kHz). The mechanisms were clearly distinguished by comparing the differences among colored fluids, suspended particles, and sinking particles. The suspended particles rotated around the antinode with a speed up to 55.1 µm s-1 at 100 Vpp by the acoustic streaming and they approached each other by the secondary acoustic radiation force. The sinking particles concentrated at the node with a speed up to 22.3 µm s-1 at 100 Vpp by bouncing on the vibrating surface and the primary acoustic radiation force. We have reproduced the classical standard/inverse Chladni patterns in an aqueous environment for the first time, and they were leveraged to separate SiO2 particles with different sizes. The big particles with an average diameter of 9.68 µm were concentrated at the node while the small particles with an average diameter of 2.72 µm were collected at the antinode within 2 min. These results not only provide insightful perspectives of basic mechanisms, but also open up new possibilities for advanced acoustic tweezers.

Droplet Microfluidic Devices: Working Principles, Fabrication Methods, and Scale-Up Applications.

Compared with the conventional emulsification method, droplets generated within microfluidic devices exhibit distinct advantages such as precise control of fluids, exceptional monodispersity, uniform morphology, flexible manipulation, and narrow size distribution. These inherent benefits, including intrinsic safety, excellent heat and mass transfer capabilities, and large surface-to-volume ratio, have led to the widespread applications of droplet-based microfluidics across diverse fields, encompassing chemical engineering, particle synthesis, biological detection, diagnostics, emulsion preparation, and pharmaceuticals. However, despite its promising potential for versatile applications, the practical utilization of this technology in commercial and industrial is extremely limited to the inherently low production rates achievable within a single microchannel. Over the past two decades, droplet-based microfluidics has evolved significantly, considerably transitioning from a proof-of-concept stage to industrialization. And now there is a growing trend towards translating academic research into commercial and industrial applications, primarily driven by the burgeoning demands of various fields. This paper comprehensively reviews recent advancements in droplet-based microfluidics, covering the fundamental working principles and the critical aspect of scale-up integration from working principles to scale-up integration. Based on the existing scale-up strategies, the paper also outlines the future research directions, identifies the potential opportunities, and addresses the typical unsolved challenges.

Acoustofluidic one-step production of plasmonic Ag nanoparticles for portable paper-based ultrasensitive SERS detection of bactericides.

SERS measurements for monitoring bactericides in dairy products are highly desired for food safety problems. However, the complicated preparation process of SERS substrates greatly impedes the promotion of SERS. Here, we propose acoustofluidic one-step synthesis of Ag nanoparticles on paper substrates for SERS detection. Our method is economical, fast, simple, and eco-friendly. We adopted laser cutting to cut out appropriate paper shapes, and aldehydes were simultaneously produced at the cutting edge in the pyrolysis of cellulose by laser which were leveraged as the reducing reagent. In the synthesis, only 5 µL of Ag precursor was added to complete the reaction, and no reducing agent was used. Our recently developed acoustofluidic device was employed to intensely mix Ag+ ions and aldehydes and spread the reduced Ag nanoparticles over the substrate. The SERS substrate was fabricated in 1 step and 3 min. The standard R6G solution measurement demonstrated the excellent signal and prominent uniformity of the fabricated SERS substrates. SERS detection of the safe concentration of three bactericides, including tetracycline hydrochloride, thiabendazole, and malachite green, from food samples can be achieved using fabricated substrates. We take the least cost, time, reagents, and steps to fabricate the SERS substrate with satisfying performance. Our work has an extraodinary meaning for the green preparation and large-scale application of SERS.

Acousto-dielectric tweezers for size-insensitive manipulation and biophysical characterization of single cells.

The intrinsic biophysical properties of cells, such as mechanical, acoustic, and electrical properties, are valuable indicators of a cell's function and state. However, traditional single-cell biophysical characterization methods are hindered by limited measurable properties, time-consuming procedures, and complex system setups. This study presents acousto-dielectric tweezers that leverage the balance between controllable acoustophoretic and dielectrophoretic forces applied on cells through surface acoustic waves and alternating current electric fields, respectively. Particularly, the balanced acoustophoretic and dielectrophoretic forces can trap cells at equilibrium positions independent of the cell size to differentiate between various cell-intrinsic mechanical, acoustic, and electrical properties. Experimental results show our mechanism has the potential for applications in single-cell analysis, size-insensitive cell separation, and cell phenotyping, which are all primarily based on cells' intrinsic biophysical properties. Our results also show the measured equilibrium position of a cell can inversely determine multiple biophysical properties, including membrane capacitance, cytoplasm conductivity, and acoustic contrast factor. With these features, our acousto-dielectric tweezing mechanism is a valuable addition to the resources available for biophysical property-based biological and medical research.

In vitro degradation behavior of silica nanoparticles under physiological conditions.

Understanding the degradability of silica nanoparticles is significant for the rational design of desired nanomaterials for various biomedical applications. However, the effect of the intrinsic properties of silica nanoparticles, such as particle shape, surface chemistry, and porosity, on kinetic degradation process under different extrinsic conditions has still received little attention. Herein, mesoporous silica nanoparticles (MSNs) with different aspect ratios (ARs, 1, 2, and 4), the corresponding PEG-functionalized MSNs, and amorphous Stöber spherical silica nanoparticles were specially designed and their degradation was evaluated in in vitro simulated physiological media. The results show that shape, surface properties and porosity of nanoparticles, as well as the component of simulated physiological media, play important roles in tuning the degradation kinetics and behaviors. Sphere-shaped MSNs have a faster degradation rate than rod-shaped counterparts. Naked MSNs are eroded from particle external surface, while PEGylated MSNs from interior of particles. And spherical MSNs display more extensive degradation than amorphous silica nanoparticles. The presence of fetal bovine serum (FBS) in Dulbecco's Modified Eagle's Medium (DMEM) can accelerate the degradation process. These results can provide useful guidelines for the rational design of silica nanoparticles for biomedical applications.

Overcoming multidrug resistance with mesoporous silica nanorods as nanocarrier of doxorubicin.

Multidrug resistance (MDR) is a major obstacle to the effective chemotherapy in many human malignancies. Nanoparticulate drug delivery systems (NDDSs) have been reported to be able to bypass MDR, but the cancer therapeutic efficacy is still limited. In this study, we firstly designed the nonspherical mesoporous silica nanorods (MSNRs) with aspect ratio (AR) of 1.5 and 5 as drug delivery systems of doxorubicin to overcome multidrug resistance. For drug loading, the long-rod MSNRs (NLR, AR = 5) showed higher drug loading capacity of doxorubicin (DOX) than the short-rod MSNRs (NSR, AR = 1.5). NLR encapsulated DOX had increased intracellular DOX accumulation in drug-resistant Chinese hamster ovary (CHO) cells compared with free DOX by observablly increased cellular uptake and significantly prolonged intracellular drug retention. It further exhibited increased cytotoxicity compared with free DOX under different drug concentrations. These findings may provide a new perspective for designing high-performance nanoparticulate drug delivery systems for bypassing multidrug resistance of cancer therapy.

Acoustics-Actuated Microrobots.

Microrobots can operate in tiny areas that traditional bulk robots cannot reach. The combination of acoustic actuation with microrobots extensively expands the application areas of microrobots due to their desirable miniaturization, flexibility, and biocompatibility features. Herein, an overview of the research and development of acoustics-actuated microrobots is provided. We first introduce the currently established manufacturing methods (3D printing and photolithography). Then, according to their different working principles, we divide acoustics-actuated microrobots into three categories including bubble propulsion, sharp-edge propulsion, and in-situ microrotor. Next, we summarize their established applications from targeted drug delivery to microfluidics operation to microsurgery. Finally, we illustrate current challenges and future perspectives to guide research in this field. This work not only gives a comprehensive overview of the latest technology of acoustics-actuated microrobots, but also provides an in-depth understanding of acoustic actuation for inspiring the next generation of advanced robotic devices.

Acoustofluidics for simultaneous nanoparticle-based drug loading and exosome encapsulation.

Nanocarrier and exosome encapsulation has been found to significantly increase the efficacy of targeted drug delivery while also minimizing unwanted side effects. However, the development of exosome-encapsulated drug nanocarriers is limited by low drug loading efficiencies and/or complex, time-consuming drug loading processes. Herein, we have developed an acoustofluidic device that simultaneously performs both drug loading and exosome encapsulation. By synergistically leveraging the acoustic radiation force, acoustic microstreaming, and shear stresses in a rotating droplet, the concentration, and fusion of exosomes, drugs, and porous silica nanoparticles is achieved. The final product consists of drug-loaded silica nanocarriers that are encased within an exosomal membrane. The drug loading efficiency is significantly improved, with nearly 30% of the free drug (e.g., doxorubicin) molecules loaded into the nanocarriers. Furthermore, this acoustofluidic drug loading system circumvents the need for complex chemical modification, allowing drug loading and encapsulation to be completed within a matter of minutes. These exosome-encapsulated nanocarriers exhibit excellent efficiency in intracellular transport and are capable of significantly inhibiting tumor cell proliferation. By utilizing physical forces to rapidly generate hybrid nanocarriers, this acoustofluidic drug loading platform wields the potential to significantly impact innovation in both drug delivery research and applications.

Acoustofluidic black holes for multifunctional in-droplet particle manipulation.

Acoustic black holes offer superior capabilities for slowing down and trapping acoustic waves for various applications such as metastructures, energy harvesting, and vibration and noise control. However, no studies have considered the linear and nonlinear effects of acoustic black holes on micro/nanoparticles in fluids. This study presents acoustofluidic black holes (AFBHs) that leverage controlled interactions between AFBH-trapped acoustic wave energy and particles in droplets to enable versatile particle manipulation functionalities, such as translation, concentration, and patterning of particles. We investigated the AFBH-enabled wave energy trapping and wavelength shrinking effects, as well as the trapped wave energy-induced acoustic radiation forces on particles and acoustic streaming in droplets. This study not only fills the gap between the emerging fields of acoustofluidics and acoustic black holes but also leads to a class of AFBH-based in-droplet particle manipulation toolsets with great potential for many applications, such as biosensing, point-of-care testing, and drug screening.

Acoustofluidic multimodal diagnostic system for Alzheimer's disease.

Alzheimer's disease (AD) is a progressive and irreversible neurodegenerative brain disorder that affects tens of millions of older adults worldwide and has significant economic and societal impacts. Despite its prevalence and severity, early diagnosis of AD remains a considerable challenge. Here we report an integrated acoustofluidics-based diagnostic system (ADx), which combines triple functions of acoustics, microfluidics, and orthogonal biosensors for clinically accurate, sensitive, and rapid detection of AD biomarkers from human plasma. We design and fabricate a surface acoustic wave-based acoustofluidic separation device to isolate and purify AD biomarkers to increase the signal-to-noise ratio. Multimodal biosensors within the integrated ADx are fabricated by in-situ patterning of the ZnO nanorod array and deposition of Ag nanoparticles onto the ZnO nanorods for surface-enhanced Raman scattering (SERS) and electrochemical immunosensors. We obtain the label-free detections of SERS and electrochemical immunoassay of clinical plasma samples from AD patients and healthy controls with high sensitivity and specificity. We believe that this efficient integration provides promising solutions for the early diagnosis of AD.

A solution to the biophysical fractionation of extracellular vesicles: Acoustic Nanoscale Separation via Wave-pillar Excitation Resonance (ANSWER).

High-precision isolation of small extracellular vesicles (sEVs) from biofluids is essential toward developing next-generation liquid biopsies and regenerative therapies. However, current methods of sEV separation require specialized equipment and time-consuming protocols and have difficulties producing highly pure subpopulations of sEVs. Here, we present Acoustic Nanoscale Separation via Wave-pillar Excitation Resonance (ANSWER), which allows single-step, rapid (<10 min), high-purity (>96% small exosomes, >80% exomeres) fractionation of sEV subpopulations from biofluids without the need for any sample preprocessing. Particles are iteratively deflected in a size-selective manner via an excitation resonance. This previously unidentified phenomenon generates patterns of virtual, tunable, pillar-like acoustic field in a fluid using surface acoustic waves. Highly precise sEV fractionation without the need for sample preprocessing or complex nanofabrication methods has been demonstrated using ANSWER, showing potential as a powerful tool that will enable more in-depth studies into the complexity, heterogeneity, and functionality of sEV subpopulations.