Droplet Trapping Research Articles

Acoustically tunable intra-droplet assembly of organoids towards high-throughput tumor model construction

Three-dimensional cell organoids and spheroids are increasingly recognized as physiologically relevant models for ex vivo research and therapeutic screening. However, there remains a need for a cost-effective and convenient platform to fabricate these models with well-defined architectures for downstream applications. Here, this work proposes micro models constructing in the droplet platform, by acoustic forces, that can obtain 3D cellular organoids with tunable structures at high yield. By modulating signals applied to acoustic devices, a periodic potential array with adjustable structures is formed, enabling uniform trapping and patterning of droplets. Simultaneously, the localized potential is also formed inside droplets, resulting in the active assembly of cells. Then, hydrogel droplets containing the constructed models are crosslinked, providing ECM to mimic the 3D scaffolds. In experiment, MCF-7 cells are assembled into spherical, linear and cross structures. This method can generate thousands of tumor spheroids in one minute. The long-term culturing results indicate an enhanced efficiency in forming mature tumor models compared with control groups. This biomanufacturing technology has the advantages of being high-throughput, low-cost, and tunable, making it a robust tool for droplet-based applications such as tissue engineering, drug screening, and disease modeling etc.

Three-dimensional isotropic imaging of live suspension cells enabled by droplet microvortices

Fast, nondestructive three-dimensional (3D) imaging of live suspension cells remains challenging without substrate treatment or fixation, precluding scalable single-cell morphometry with minimal alterations. While optical sectioning techniques achieve 3D live cell imaging, lateral versus depth resolution differences further complicate analysis. We present a scalable microfluidic method capable of 3D fluorescent isotropic imaging of live, nonadherent cells suspended inside picoliter droplets with high-speed single-cell volumetric readout (800 to 1,200 slices in 5 to 8 s) and near-diffraction limit resolution (~216 nm). The platform features a droplet trap array that leverages flow-induced droplet interfacial shear to generate intradroplet microvortices, which rotate single cells on their axis to enable optical projection tomography (OPT)-based imaging. This allows gentle (~1 mPa shear stress) observation of cells encapsulated inside nontoxic isotonic buffer droplets, facilitating scalable OPT acquisition by simultaneous spinning of hundreds of cells. We demonstrate 3D imaging of live myeloid and lymphoid cells in suspension, including K562 cells, as well as naive and activated T cells-small cells prone to movement in their suspended phenotype. Our fully suspended, orientation-independent cell morphometry, driven by isotropic imaging and spherical harmonic analysis, enabled the study of primary T cells across various immunological activation states. This approach unveiled six distinct nuclear content distributions, contrasting with conventional 2D images that typically portray spheroid and bean-like nuclear shapes associated with lymphocytes. Our arrayed-droplet OPT technology is capable of isotropic, single live-cell 3D imaging, with the potential to perform large-scale morphometry of immune cell effector function states while providing compatibility with microfluidic droplet operations.

The Cryptochrome CryA Regulates Lipid Droplet Accumulation, Conidiation, and Trap Formation via Responses to Light in Arthrobotrys oligospora.

Light is a key environmental factor affecting conidiation in filamentous fungi. The cryptochrome/photolyase CryA, a blue-light receptor, is involved in fungal development. In the present study, a homologous CryA (AoCryA) was identified from the widely occurring nematode-trapping (NT) fungus Arthrobotrys oligospora, and its roles in the mycelial growth and development of A. oligospora were characterized using gene knockout, phenotypic comparison, staining technique, and metabolome analysis. The inactivation of AocryA caused a substantial decrease in spore yields in dark conditions but did not affect spore yields in the wild-type (WT) and ∆AocryA mutant strains in light conditions. Corresponding to the decrease in spore production, the transcription of sporulation-related genes was also significantly downregulated in dark conditions. Contrarily, the ∆AocryA mutants showed a substantial increase in trap formation in dark conditions, while the trap production and nematode-trapping abilities of the WT and mutant strains significantly decreased in light conditions. In addition, lipid droplet accumulation increased in the ∆AocryA mutant in dark conditions, and the mutants showed an increased tolerance to sorbitol, while light contributed to the synthesis of carotenoids. Finally, AoCryA was found to affect secondary metabolic processes. These results reveal, for the first time, the function of a homologous cryptochrome in NT fungi.

Biomimetic Fog Collector with Hybrid and Gradient Wettabilities.

Water scarcity is a global problem and collecting water from the air is a viable solution to this crisis. Inspired by Namib Desert beetle, leaf venation and spider silk, we designed an integrated biomimetic system with hybrid wettability and wettability gradient. The hybrid hydrophilic-hydrophobic wettability design that bionomics desert beetle's back can construct a three-dimensional topography with a water layer on the surface, expanding the contact area with the fog flow and thus improving the droplet trapping efficiency. The venation-like structure with wettability gradient not only provides a planned path for water transportation, but also accelerates water removal under the synergistic effect of gravity and wettability driving force, thus further improving the surface regeneration rate. The collector combines droplet capture, coalescence, transportation, separation, and storage capabilities, which provides new ideas for the design of future high-efficiency fog collectors.

Trapping of isotropic droplets by disclinations in nematic liquid crystals controlled by surface anchoring and elastic constant disparity.

Linear defects such as dislocations and disclinations in ordered materials attract foreign particles since they replace strong elastic distortions at the defect cores. In this work, we explore the behavior of isotropic droplets nucleating at singular disclinations in a nematic liquid crystal, predesigned by surface photopatterning. Experiments show that in the biphasic nematic-isotropic region, although the droplets are attracted to the disclination cores, their centers of mass shift away from the core centers as the temperature increases. The shift is not random, being deterministically defined by the surrounding director field. The effect is explained by the balance of interfacial anchoring and bulk elasticity. An agreement with the experiment can be achieved only if the model accounts for the disparity of the nematic elastic constants; the so-called one-constant approximation, often used in the theoretical analysis of liquid crystals, produces qualitatively wrong predictions. In particular, the experimentally observed shift towards the bend region around a +1/2 disclination core can be explained only when the bend constant is larger than the splay constant. The described dependence of the precise location of a foreign inclusion at defect cores on the elastic and surface anchoring properties can be used in rational design of microscale architectures.

Characteristics of droplet dispersion and interpersonal exposure in indoor and outdoor dining areas of urban street: Elementary numerical investigations

Indoor and outdoor dining areas are special spaces where people generally sit close and talk face-to-face for a long time (∼hour). Semi-open dining area is of popularity among them, one of the reason is the thoughts that outdoor dining area have experienced small infectious risks, even though this opinion has not been thoroughly validated yet. As a novelty, this study conducts CFD simulations to investigate droplet dispersion and face-to-face exposure risk of customers in the indoor and outdoor dining areas of typical street canyons (H/W = 1) by coupling indoor/outdoor airflows. Different scenarios are considered including index patient locations, the setting of the desk isolation board (DIB, HDIB = 0.6 m or 1.2 m), and the outdoor semi-open street roof (SSR).Results show that indoor dining spaces with single-sided ventilation experience one-order poorer ventilation (net escape velocity NEV∼0.036 m/s) and subsequently 10–200 times higher face-to-face droplet trap fraction than those in outdoor dining areas (NEV∼0.407 m/s, TF∼3–102 ppm). Indoor desk isolation board can decrease face-to-face TF by 50 %, but in outdoors, TF without desk-isolation board is 6.5 ppm for 50 μm droplet and this value changes little or slightly increases as HDIB is 0.6 m (∼8.9 ppm) or 1.2 m (∼7.7 ppm), indicating DIB seems not helpful to reduce airborne exposure risk in outdoors. Moreover, the setting of an outdoor semi-open street roof (SSR) may reduce NEV in the indoor or outdoor dining area by half or 90 % respectively. Therefore, SSR may raise indoor and outdoor customers' face-to-face exposure risk (e.g. TF) by hundreds of times and seventy times respectively.

Simple Method for On-Demand Droplet Trapping in a Microfluidic Device Based on the Concept of Hydrodynamic Resistance.

We demonstrate an innovative method to catch the desired droplets from a train of droplets and immobilize them in traps located in an integrated microfluidic device. To this end, water-in-oil droplets are generated in a flow-focusing junction and then guided to a channel connected to chambers designated for on-demand droplet trapping. Each chamber is connected to a side channel through a batch of microposts. The side channels are also connected to the flexible poly(vinyl chloride) tubes, which can be closed by attaching binder clips. The hydrodynamic resistance of the routes in the device can be changed by opening and closing the binder clips. In this way, droplets are easily guided into individual traps based on the user's demand. A set of numerical simulations was also conducted to investigate the authenticity of the employed idea and to find the optimal geometry for implementing our strategy. This simple method can be easily employed for on-demand droplet trapping without using on-chip valves or complex off-chip actuators proposed in previous studies.

Controlled Trapping and Release of Droplets through Dynamically Reconfigurable Surfaces

AbstractMost previous publications attempt to control the trapping and release of liquid droplets on a surface through tuning the surface energy of the substrate, for example, by photoisomerization. In this work, the energy barrier between the trapped and released state is raised or lowered by controlling the droplet contact area. This is achieved by designing a dynamically reconfigurable surface where an external stimulus is used to change the surface topography. The system consists of two microstructured components, one being ridges separated by microtrenches and the other a perforated membrane. They are aligned in such a way that the ridges can be raised or lowered through the perforations with the help of a piezo drive. This way the topography changes from surfaces consisting of hills to one consisting of holes – and back again. This dynamic surface reconfiguration changes the contact area such that drops resting or rolling on them become captured or released on demand.

Solid-like slippery surface for anti-icing and efficient fog collection

Ice accumulation on the surface of aerospace and transportation equipment under extreme weather seriously endangers the safety and efficiency of equipment operation. In recent years, metal corrosion, especially in the marine engineering industry, has become a global concern. The slippery liquid infused porous surface (SLIPS) is a new bionic surface which has caused extensive research in recent years, and has great potential in anti-corrosion and anti-icing. In this study, a multifunctional solid-like lubricated slippery surface (SL-SLIPS) was developed by superhydrophobic modification of the base of copper hydroxide nanoneedles and injection of solid-like lubricant synthesized by embedding oil-stored mesoporous silica nanoparticles on the surface of epoxy resin. At −30 °C, the delayed icing time on the surface of the SL-SLIPS sample was 22.6 times that of the original copper substrate. The ice adhesion was tested on the surface of the sample, and the results showed that the adhesion of the icicle on the SL-SLIPS surface was the smallest, which decreased by 89% compared with the original copper substrate. In addition, SL-SLIPS have excellent corrosion resistance and rapid droplet trapping, nucleation and drop. It has good practical application prospects in corrosion resistance of marine equipment and icing resistance of aerospace and transportation equipment. This study provides new ideas for the design of new slippery surfaces and their application in practical fields, such as anti-corrosion of offshore equipment, anti-icing of equipment in aviation and fog collection engineering in industry.

Effects of ultrasound on the removal of emulsion plugging in oil reservoirs

The generation of water-in-crude oil emulsions in a reservoir can cause formation damage due to droplet trapping at pore spaces. The removal of the damage is anticipated to be inexpensive and eco-friendly when done with ultrasound as opposed to chemical demulsifiers. The influence of ultrasonic power and frequencies on the removal process, however, is not well understood. Additionally, the process's underlying mechanism is largely speculated. In this study, the effect of ultrasound on the removal of emulsion plugging in oil reservoirs was investigated using a glass micromodel. Emulsion blockage during oil production was replicated in the micromodel and subjected to different ultrasonic frequencies (20 and 40 kHz) and powers (100–1000 watt). The experiments demonstrate that when ultrasonic power and frequency increase from 100 to 1000 watt and 20–40 kHz, respectively, demulsification effectiveness decreases. Ultrasound at low frequency (20 kHz) and power (100 watt) proved to be the most efficient condition to dislodge trapped emulsions in the micromodel pores, facilitate droplet coalescence and increase fluid recovery. The percentage of recovered emulsions increased to 58 % when the micromodel was exposed to ultrasound (20 kHz, 100 watt), as opposed to 53.3 % in the case without ultrasound. This study provides insights into the microscopic behavior of emulsions under the influence of ultrasonic waves, allowing petroleum engineers to optimize ultrasound demulsification process.

Co-stabilization mechanisms of solid particles and soluble compounds in hybrid Pickering emulsions stabilized by unrefined apple pomace powder

Vegetal by-product powders, such as apple pomace powder, represent an efficient green alternative to conventional emulsifiers to stabilize oil-in-water Pickering emulsions. The natural material being used without purification, its multiple components shall impact differently the emulsion stability and interact with each other. These effects might be either synergic, competitive, or both depending on their ratio. Herein, the co-stabilizing effects of the apple pomace compounds on Pickering emulsions were explored from microscopic to macroscopic scales. The apple pomace powder was first fragmented into several fractions which were characterized (particle sizes, shape and wettability, oil-water surface activity). Emulsions were then prepared by varying the stabilizing material composition using the different fractions obtained from the apple powder. Emulsions properties (droplet size, rheological properties) and stability were monitored over a month. The key role of solid particles in the surface activity of the apple pomace powder was shown by interfacial tension measurements. All particle-stabilized emulsions were stable against coalescence during storage despite high droplet diameters (up to 80 μm) and low surface coverage. Soluble matter and particles show a synergic behavior to stabilize oil-in-water emulsions. The soluble matter drives droplet size whereas insoluble cellulosic particles are responsible for emulsion stability by both Pickering adsorption to the oil droplet and oil droplet trapping in a three-dimensional network. This study provides new insights into the understanding of natural powders stabilizing mechanisms that may lead to a better prediction of the potential of vegetal by-products as stabilizing agents from their chemical composition.

Mechanism of droplet motion in the typical micro-channel of porous media

The investigation of a two-phase flow in porous media has significant implications for a wide range of applications. Previous research has focused on exploring the variations in flow and phase fields in a two-phase flow using experimental and numerical methods. However, the complex structure of porous media introduces many uncertainties that can impact research outcomes. In recent years, some scholars have tried to study the dynamic mechanics of a two-phase flow through typical structures to eliminate these confounding factors. Therefore, this paper focuses on examining the flow patterns of dispersed phases with different sizes during the displacement process based on the typical micro-channel of porous media. Furthermore, the study examines various dimensionless parameters that impact alterations in the streamlines of a two-phase flow as well as the carrying capacity for dispersed phases. The findings suggest that the capillary number governs the ability of the continuous phase to transport the droplet. Consequently, the dispersed droplets tend to become trapped in weak flow regions. The dynamic mechanisms of the dispersive droplet trapping are systematically analyzed by combining the numerical simulation results and experimental evidence from previous studies. Based on these findings, the paper puts forth some mechanistic suggestions that could contribute to a more effective displacement of a two-phase flow in porous media.

On-demand light-driven release of droplets stabilized via a photoresponsive fluorosurfactant

Water-in-oil droplets have emerged as promising microreactors for high-throughput biochemical analysis due to their features of reduced sample consumption and automated operation. For a typical screening application, droplets are often trapped for continuous monitoring of the reaction over an extended period, followed by the selective retrieval of targeted droplets based on the after-effect of biochemical reactions. While techniques for droplet trapping are well developed, retrieval of targeted droplets mainly demands complicated device fabrication or sophisticated control. Herein, facile and rapid selective droplet release is achieved by utilizing a new class of photoresponsive fluorosurfactant based on plasmonic nanoparticles. The intense photothermal response provided by this novel photoresponsive fluorosurfactant is capable of vaporizing the fluorocarbon oil at the droplet interface under laser illumination, resulting in a bubble releasing a trapped droplet on demand. A fully automated fluorescence-activated droplet release platform has also been developed to demonstrate its potential for droplet-based large-scale screening applications.

Tin(IV) Oxide Electron Transport Layer via Industrial-Scale Pulsed Laser Deposition for Planar Perovskite Solar Cells.

Electron transport layers (ETL) based on tin(IV) oxide (SnO2) are recurrently employed in perovskite solar cells (PSCs) by many deposition techniques. Pulsed laser deposition (PLD) offers a few advantages for the fabrication of such layers, such as being compatible with large scale, patternable, and allowing deposition at fast rates. However, a precise understanding of how the deposition parameters can affect the SnO2 film, and as a consequence the solar cell performance, is needed. Herein, we use a PLD tool equipped with a droplet trap to minimize the number of excess particles (originated from debris) reaching the substrate, and we show how to control the PLD chamber pressure to obtain surfaces with very low roughness and how the concentration of oxygen in the background gas can affect the number of oxygen vacancies in the film. Using optimized deposition conditions, we obtained solar cells in the n-i-p configuration employing methylammonium lead iodide perovskite as the absorber layer with power conversion efficiencies exceeding 18% and identical performance to devices having the more typical atomic layer deposited SnO2 ETL.

AoMae1 Regulates Hyphal Fusion, Lipid Droplet Accumulation, Conidiation, and Trap Formation in Arthrobotrys oligospora.

Malate dehydrogenase (MDH) is a key enzyme in the tricarboxylic acid (TCA) cycle and is essential for energy balance, growth, and tolerance to cold and salt stresses in plants. However, the role of MDH in filamentous fungi is still largely unknown. In this study, we characterized an ortholog of MDH (AoMae1) in a representative nematode-trapping (NT) fungus Arthrobotrys oligospora via gene disruption, phenotypic analysis, and nontargeted metabolomics. We found that the loss of Aomae1 led to a weakening of MDH activity and ATP content, a remarkable decrease in conidia yield, and a considerable increase in the number of traps and mycelial loops. In addition, the absence of Aomae1 also caused an obvious reduction in the number of septa and nuclei. In particular, AoMae1 regulates hyphal fusion under low nutrient conditions but not in nutrient-rich conditions, and the volumes and sizes of the lipid droplets dynamically changed during trap formation and nematode predation. AoMae1 is also involved in the regulation of secondary metabolites such as arthrobotrisins. These results suggest that Aomae1 has an important role in hyphal fusion, sporulation, energy production, trap formation, and pathogenicity in A. oligospora. Our results enhance the understanding of the crucial role that enzymes involved in the TCA cycle play in the growth, development, and pathogenicity of NT fungi.

Single Beam Acoustical Tweezers Based on Focused Beams: A Numerical Analysis of Two-Dimensional and Three-Dimensional Trapping Capabilities

Selective single beam tweezers open tremendous perspectives in microfluidics and microbiology for the micromanipulation, assembly, and mechanical properties testing of microparticles, cells, and microorganisms. In optics, single beam optical tweezers rely on tightly focused laser beams, generating a three-dimensional (3D) trap at the focal point. In acoustics, 3D traps have so-far only been reported experimentally with specific wavefields called acoustical vortices. Indeed, many types of particles are expelled (not attracted to) the center of a focused beam. Yet the trapping capabilities of focused beams have so far only been partially explored. In this paper, we numerically explore with an angular spectrum code the trapping capabilities of focused beams on a wide range of parameters (size over wavelength ratio and type of particles). We demonstrate that (i) 3D trapping of particles, droplets, and microorganisms more compressible than the surrounding fluid is possible in and beyond the Rayleigh regime [e.g., polydimethylsiloxane (PDMS), olive oil, benzene, and lipid sphere] and that (ii) 2D trapping (without an axial trap) of particles with positive contrast factor can be achieved by using the particles resonances. The 3D trapping of biocompatible functionalized PDMS particles with integrable, high-frequency focused-beam tweezers opens the way towards acoustic force spectroscopy in some force ranges that were not accessible with optical methods.

Generation of 3D Spheroids Using a Thiol-Acrylate Hydrogel Scaffold to Study Endocrine Response in ER+ Breast Cancer.

Culturing cancer cells in a three-dimensional (3D) environment better recapitulates in vivo conditions by mimicking cell-to-cell interactions and mass transfer limitations of metabolites, oxygen, and drugs. Recent drug studies have suggested that a high rate of preclinical and clinical failures results from mass transfer limitations associated with drug entry into solid tumors that 2D model systems cannot predict. Droplet microfluidic devices offer a promising alternative to grow 3D spheroids from a small number of cells to reduce intratumor heterogeneity, which is lacking in other approaches. Spheroids were generated by encapsulating cells in novel thiol–acrylate (TA) hydrogel scaffold droplets followed by on-chip isolation of single droplets in a 990- or 450-member trapping array. The TA hydrogel rapidly (∼35 min) polymerized on-chip to provide an initial scaffold to support spheroid development followed by a time-dependent degradation. Two trapping arrays were fabricated with 150 or 300 μm diameter traps to investigate the effect of droplet size and cell seeding density on spheroid formation and growth. Both trapping arrays were capable of ∼99% droplet trapping efficiency with ∼90% and 55% cellular encapsulation in trapping arrays containing 300 and 150 μm traps, respectively. The oil phase was replaced with media ∼1 h after droplet trapping to initiate long-term spheroid culturing. The growth and viability of MCF-7 3D spheroids were confirmed for 7 days under continuous media flow using a customized gravity-driven system to eliminate the need for syringe pumps. It was found that a minimum of 10 or more encapsulated cells are needed to generate a growing spheroid while fewer than 10 parent cells produced stagnant 3D spheroids. As a proof of concept, a drug susceptibility study was performed treating the spheroids with fulvestrant followed by interrogating the spheroids for proliferation in the presence of estrogen. Following fulvestrant exposure, the spheroids showed significantly less proliferation in the presence of estrogen, confirming drug efficacy.

A new mechanism of viscoelastic fluid for enhanced oil recovery: Viscoelastic oscillation

This report summarizes our recent experimental findings [Xie et al., Phys. Rev. Lett., 2022] and pore-scale simulation results [Xie et al., Phys. Rev. Fluids., 2020] on viscoelastic oscillation, which is a new observation of viscoelastic instability in the multiphase flow state. The viscoelastic oscillation causes trapping of droplets in contraction-expansion micro-channels regardless of the injection rate. Based on the force balance analysis on the viscous, capillary and elastic forces, the oscillation amplitude is found to linearly increase with viscoelasticity, and the trapped droplet size is determined by the elasto-capillary number. The oscillation also helps to extract droplets from their originally trapped positions such as dead-ends once a critical Deborah number is reached. These results successfully explain the phenomenon that the alternative injection of viscoelastic and inelastic fluids continually produces additional oil, indicating that the viscoelastic oscillation is a new important mechanism of viscoelastic fluid for enhanced oil recovery. Cited as: Xie, C., Xu, K., Qi, P., Xu, J., Balhoff, M. T. A new mechanism of viscoelastic fluid for enhanced oil recovery: Viscoelastic oscillation. Advances in Geo-Energy Research, 2022, 6(3): 267-268. https://doi.org/10.46690/ager.2022.03.10

Trapping of Aqueous Droplets under Surface Acoustic Wave-Driven Streaming in Oil-Filled Microwells.

Microwell arrays are ideal platforms for cell culturing, cell separation, and low-volume liquid handling. The ability to manipulate droplets in microwells could open up the opportunity for developing new biochemical assays. Here, we study the trapping of aqueous droplets in an oil-filled microwell driven by the application of nanometer amplitude vibrations called surface acoustic waves (SAW). We elucidate the dynamics of the droplet within the vortex toward the final trapping location and the physics of the trapping phenomenon using a theoretical model by considering the relevant forces. Our study revealed that the combined effect of acoustic radiation and hydrodynamic forces leads to droplet migration and trapping. We demarcate the trapping and nontrapping regimes in terms of the minimum critical input power required for the trapping of droplets of different sizes and densities. We find that the critical power varies as the square of the droplet size and is higher for a denser droplet. The effects of input power and droplet size on the trapping location and trapping time are also studied.

Transport of condensing droplets in Taylor-Green vortex flow in the presence of thermal noise.

We study the role of phase change and thermal noise in particle transport in turbulent flows. We employ a toy model to extract the main physics: Condensing droplets are modelled as heavy particles which grow in size, the ambient flow is modelled as a two-dimensional Taylor-Green flow consisting of an array of vortices delineated by separatrices, and thermal noise are modelled as uncorrelated Gaussian white noise. In general, heavy inertial particles are centrifuged out of regions of high vorticity and into regions of high strain. In cellular flows, we find, in agreement with earlier results, that droplets with Stokes numbers smaller than a critical value, St<St_{cr}, remain trapped in the vortices in which they are initialized, while larger droplets move ballistically away from their initial positions by crossing separatrices. We independently vary the Péclet number Pe characterizing the amplitude of thermal noise and the condensation rate Π to study their effects on the critical Stokes number for droplet trapping, as well as on the final states of motion of the droplets. We find that the imposition of thermal noise, or of a finite condensation rate, allows droplets of St<St_{cr} to leave their initial vortices. We find that the effects of thermal noise become negligible for growing droplets and that growing droplets achieve ballistic motion when their Stokes numbers become O(1). We also find an intermediate regime prior to attaining the ballistic state, in which droplets move diffusively away from their initial vortices in the presence of thermal noise.