Aqueous Droplets Research Articles

Manipulation of Cancer Cell Spheroids by Photovoltaic Tweezers: Determination of Their Charge State

Photovoltaic optoelectronic tweezers (PVOT), based on the bulk photovoltaic effect presented by certain ferroelectric crystals, have emerged as a versatile tool to manipulate and/or trap micro‐ and nano‐objects. Very recently, an ingenious method has enabled to extend PVOT manipulation to aqueous solution droplets opening the door to applications in optofluidics and biotechnology. The photovoltaic crystal is placed on the bottom of a cuvette filled with paraffin oil while the droplet is hanging from the air–oil interphase where it is manipulated. Herein, this method is used to investigate the feasibility of manipulating hybrid droplets as carriers containing a cancer multicellular spheroid inside cell culture medium. Droplets of two cancer cell lines, MCF‐7 and U‐87 MG, have been successfully manipulated by light‐induced electrical fields generated in the photovoltaic platform. From the droplet dynamics, which includes attractive and repulsive behaviors from the illuminated crystal region, the charge state of the cell culture medium and the spheroid are investigated. The results show that they are positively and negatively charged, respectively. Moreover, analyzing the migration kinetics of water, cell culture medium, and hybrid droplets, to the light of a proposed theoretical model, the charge density of both cell line spheroids has been estimated.

Binary imidazolium based aqueous ionic liquid droplet on a rough surface: a molecular dynamics study

Ionic liquids (ILs) have garnered immense attention due to their tunable physicochemical properties. In this study, we delve into the wetting behaviour and interfacial properties of aqueous binary IL mixtures containing [EMIM] [BF4] and [HMIM] [BF4] on both smooth and rough graphite surfaces. Employing classical molecular dynamics simulations, we systematically explored the effects of IL concentration and surface morphology on wetting phenomena. To ensure the equilibrium of the systems, we began by calculating the interaction energies between the components. Subsequently, contact angles were computed, shedding light on the wetting characteristics of the aqueous binary IL mixtures. We meticulously examined the density profiles of the IL systems from the substrate to unravel the dominance of one IL over the other, exploiting the hydrophilic nature of [EMIM] [BF4] and the hydrophobic nature of [HMIM] [BF4]. A comprehensive analysis of hydrogen bond distribution allowed us to discern the intricate nature of these aqueous binary IL droplets. We probed the changes in intermolecular forces within the aqueous IL solution by investigating surface tensions. The tunability of these ILs emerges as a significant aspect of our study, paving the way for their tailored application in engineering and scientific domains. Schematic diagram showing the density distribution of hydrophilic and hydrophobic ILs ([EMIM] [BF4] and [HMIM] [BF4]) in water and also the hydrogen bond (HB) distribution of anion ([BF4]−) water inside the droplet on a pillared surface with a surface fraction ( f ) of 0.4 and pillar height of H = 4 .

Responsive release of guest aqueous droplet in liquid crystal droplet assists the detection of organophosphorus using commercial pregnancy test strip

Here, we report a liquid crystal (LC) double-emulsion based system for organophosphorus (OPs) sensing by controllable release of guest aqueous droplets (GADs) cargo. First, double liquid crystal emulsion, water in liquid crystal in water (W/LC/W), with size between 4∼ 20 μm was prepared with GADs loaded at the center of liquid crystal droplet (LCD) through elasticity trap. The LCDs were monodispersed and response sensitively to amphiphilic surfactant with opposite charge. With the capability of ejecting GADs with human chorionic gonadotropin (hCG) as cargo located in LCD to continuous aqueous phase, responsive vehicles are obtained and applied in the repurposing of commercial pregnancy test strip (PTS). With hydrophobic alkyl chain and hydrophilic head, myristoylcholine (Myr) is a good amphiphilic surfactant to trigger the release of hCG, and substrate for the enzymatic hydrolysis of acetylcholine esterase (AChE). Provided with the inhibition effect of OPs to the hydrolysis ability of AChE, OPs correlate positively with the concentration of Myr, thus correlate well with the release of hCG in W/LC/W emulsion. Thus, OPs residue in river or food is readily visualized using commercial PTS.

High precision acoustofluidic synthesis of stable, biocompatible water-in-water emulsions

Water-in-water (w/w) emulsions, comprising aqueous droplets within another continuous aqueous phase, rely on a low interfacial tension for stability. Thus far, it has been challenging to control their size and stability without the use of stabilizers. In this study, we introduce a microfluidic technique that addresses these challenges, producing stable w/w emulsions with precisely controlled size and uniformity. Results shows that using an acoustically actuated microfluidic mixer, PEG, Dextran, and alginate solutions (84.66 mPa.s viscosity difference) were homogenized rapidly, forming uniformly distributed w/w emulsions stabilized in alginate gels.The emulsion size, uniformity, and shear sensitivity can be tuned by modifying the alginate concentration. Biocompatibility was evaluated by monitoring the viability of kidney cells in the presence of emulsions and gels. In conclusion, this study not only showed emulsion formation with a high mixing efficiency exceeding 90 % for all viscosities, actuated at an optimized frequency of 1.064 MHz, but also demonstrated that an aqueous, solvent, and emulsifier-free composition exhibited remarkable biocompatibility, holding promise for precise drug delivery, cosmetics, and food applications.

Electrocoalescence of unequal-sized aqueous droplet pair in non-conductive medium

Electrocoalescence of unequal-sized aqueous droplet pair in non-conductive medium

Are Hydroxyl Radicals Spontaneously Generated in Unactivated Water Droplets?

Spontaneous ionization/breakup of water at the surface of aqueous droplets has been reported with evidence ranging from formation of hydrogen peroxide and hydroxyl radicals, indicated by ions at m/z 36 attributed to OH•‐H3O+ or (H2O‐OH2)+• as well as oxidation products of radical scavengers in mass spectra of water droplets formed by pneumatic nebulization. Here, aqueous droplets are formed both by nanoelectrospray, which produces highly charged nanodrops with initial diameters ~100 nm, and a vibrating mesh nebulizer, which produces 2 – 20 µm droplets that are less highly charged. The lifetimes of these droplets range from 10s of µs to 560 ms and the surface‐to‐volume ratios span ~100‐fold range. No ions at m/z 36 are detected with pure water, nor are significant oxidation products for the two radical scavengers that were previously reported to be formed in high abundance. These and other results indicate that prior conclusions about spontaneous hydroxyl radical formation in unactivated water droplets are not supported by the evidence and that water appears to be stable at droplet surfaces over a wide range of droplet size, charge and lifetime.

Valence Electronic Structure of Interfacial Phenol in Water Droplets.

Biochemistry and a large part of atmospheric chemistry occur in aqueous environments or at aqueous interfaces, where (photo)chemical reaction rates can be increased by up to several orders of magnitude. The key to understanding the chemistry and photoresponse of molecules in and "on" water lies in their valence electronic structure, with a sensitive probe being photoelectron spectroscopy. This work reports velocity-map photoelectron imaging of submicrometer-sized aqueous phenol droplets in the valence region after nonresonant (288 nm) and resonance-enhanced (274 nm) two-photon ionization with femtosecond ultraviolet light, complementing previous liquid microjet studies. For nonresonant photoionization, our concentration-dependent study reveals a systematic decrease in the vertical binding energy (VBE) of aqueous phenol from 8.0 ± 0.1 eV at low concentration (0.01 M) to 7.6 ± 0.1 eV at high concentration (0.8 M). We attribute this shift to a systematic lowering of the energy of the lowest cationic state with increasing concentration caused by the phenol dimer and aggregate formation at the droplet surface. Contrary to nonresonant photoionization, no significant concentration dependence of the VBE was observed for resonance-enhanced photoionization. We explain the concentration-independent VBE of ∼8.1 eV observed upon resonant ionization by ultrafast intermediate state relaxation and changes in the accessible Franck-Condon region as a consequence of the lowering of the intermediate state potential energy due to the formation of phenol excimers and excited phenol aggregates. Correcting for the influence of electron transport scattering in the droplets reduced the measured VBEs by 0.1-0.2 eV.

The presence of nanoparticles in aqueous droplets containing plant-derived biopolymers plays a role in heterogeneous ice nucleation.

Organic matter can initiate heterogeneous ice nucleation in supercooled water droplets, thereby influencing atmospheric cloud glaciation. Predicting the ice nucleation ability of organic matter-containing cloud droplets is challenging due to the unknown mechanism for templating ice. Here, we observed the presence of nanoparticles in aqueous samples of known ice-nucleating biopolymers cellulose and lignin, as well as in newly identified ice-nucleating biopolymers xylan and laminarin. Using our drop Freezing Ice Nuclei Counter (FINC), we measured the median ice nucleation temperature (T50) of xylan and of laminarin droplets of 2 μl to be -14.2 and -20.0 °C, respectively. Next, we characterized these samples using nanoparticle tracking analysis, and we detected and quantified nanoparticles with mean diameters between 132 and 267nm. Xylan contained the largest nanoparticles and froze at higher temperatures. Xylan also dictated the freezing in a 1:1:1:1 mixture with cellulose, lignin, laminarin, and xylan. Filtration experiments down to 300 kDa with the xylan sample indicated that the presence of nanoparticles triggered freezing. Overall, only samples with mean diameters above 150nm froze above -20 °C. Furthermore, we determined the ice-active site densities normalized to particle concentrations, surface area, and mass of the nanoparticles to show that the samples' nucleation site densities are similar to sea spray aerosols and nanometer-sized dust. The identification and characterization of xylan and laminarin as nanometer-sized ice-nucleating substances expands the growing list of organic matter capable of impacting cloud formation and thus climate.

NaClO3 Crystal Growth and Dissolution by Temperature Cycling in a Sessile Droplet

Sodium chlorate is the most popular compound used to study spontaneous symmetry breaking by means of crystallization. Therefore, it is important to know the behavior of the solid particles. NaClO3 crystal growth and dissolution are investigated in an aqueous sessile droplet subjected to numerous temperature cycles. On cooling, in addition to the classical formation of repeated elongated fluid inclusions, there is a reproducible appearance of prismatic fluid inclusions (PFIs) at the corners of single crystals. The underlying mechanism involves the complete termination of the (110) face growth and the propagation of the {100} faces, which can close the PFIs. This study reports that on heating, transient donut-like single crystals formed, which could lead to their segmentation, even without stirring the suspension. The systematic addition of other sodium salts with chlorine atoms at different oxidation states did not change these observations.

Pre-contact electrification of the aqueous droplets on a hydrophobic surface

The solid-liquid contact electrification (CE) has been widely studied due to its importance in diverse fields. However, the evidence indicates that the relevant charge transfer or the charge accumulation in the aqueous droplets might start prior to the solid-liquid contact. Nevertheless, this pre-contact electrification of the aqueous droplets is still rarely studied. Here, the effect of a charged FEP (Fluorinated ethylene propylene) hydrophobic film on the water droplets falling from a syringe needle was studied systematically. It was found that the aqueous droplets can be charged differently in the vicinity of the FEP film, shifting gradually from positively to negatively charged. Based on the kinetic analysis of the droplet falling motion and the numerical simulation of the electrostatic field in the vicinity of the FEP film, we propose a charge-transfer mechanism for this pre-contact electrification of the water droplets, which involves a competition between the positive-charge generation caused by the detachment of the aqueous droplets under the electrostatic field and the capture of the negative-charge jets by the aqueous droplets. This competitive mechanism, verified by both the aperture limiting effect and the FEP-surface-potential dependence of the negative charge jets, allows for a flexible control over the charge polarity and charge amount of the falling water droplets. The outcome of this study expands the solid-liquid charge-transfer mechanism and broadens the application scope of the aqueous droplet charging.

Pressure-driven formation of sub-femtoliter droplets in constricted T-shaped nanofluidic channel and application to isolation and counting of single nanoparticles

Nanofluidics has developed, enabling the analysis of single nanoparticles such as viruses and macromolecules using nanochannels. However, since nanoparticles migrate randomly by Brownian motion with displacements comparable to the nanochannel size, it is difficult to array and process nanoparticles individually in continuous phase flows with biological analyte concentrations of pM–nM. The present study developed a method to generate aqueous droplets of 102 aL–100 fL volumes in an organic phase in a nanochannel by pressure-driven flow control, and applied the method to the isolation of single nanoparticles with 101−102 pM concentrations. A T-shaped nanochannel with a narrow constriction section was proposed to locally enhance the shear force to form smaller droplets by 102 kPa external pressure, without increasing the pressure loss by downscaling the channel. The proposed channel design was validated, droplets with 0.8–1.0 fL volumes were formed with pL/min flow rates at Ca = 0.5–1.4 × 10−3, and the dynamics of droplet generation was well characterized by the linear scaling law. Using the developed device, encapsulation and counting of single fluorescent nanoparticles and single fluorescently-labeled DNA molecules in droplets were successfully demonstrated. This work will contribute to the development of droplet nanofluidics and ultrasmall analytical applications.

Enhancing water transportation capacity by asymmetrical patterned surface with super-wettability

Spontaneous and directional water droplet transportation based on patterned surface with super-wettability is crucial for the development of frontier science technology. However, water droplet transportation cannot meet both long distance and fast transportation simultaneously. Here, we overcame this limitation by proposing an asymmetric serial brachistochrone-shaped pattern (ASBP). Water droplet could be transported on the ASBP with a transportation distance of 72.52 mm and a transportation velocity of 158 mm/s after a series of single-factor experiments, orthogonal design optimization, and junction transition optimization. In addition, the water droplet could be transported on a curved ASBP, a super-long ASBP for multi-droplet scenarios, and an ASBP at an inclination angle. Moreover, acidic and alkaline aqueous solution droplet showed similar transportation distance and transportation velocity on the ASBP. Based on the aforementioned superb water transportation capacity, this ASBP can be applied in the fields of fog collection, solution mixing and reaction, and reagent detection. This work has strong implications for promoting the application of patterned surface with super-wettability in the field of high-performance fluid transportation systems.

Toward an Understanding of Uric Acid Particle Dissolution: Surface and Bulk-Driven Water Activity.

Uric acid particles contribute to kidney stones, and natural processes for the elimination of stones depend on solute-solvent interactions. The process of uric acid dissolution has previously been understood via the lens of solubility; however, for pure and mixed salt solutions, these approaches do not provide a comprehensive picture of nanoscale particle solution thermodynamics. Unlike solubility measurements, water activity measurements provide us with information about the chemical potential responsible for the migration of water molecules driving the dissolution of particles. In this work, we used in situ experimental tools to estimate water activity values for pure uric acid aqueous droplets at different stages of droplet growth. The process of cloud formation, i.e., water condensation on a solute particle resulting in aqueous droplet formation, was leveraged to compare the water affinity for nanosized uric acid particles with a well-studied inorganic salt, sodium chloride. Specifically, we investigated microscopic uric acid particles (nanoparticles <300 nm, amorphous and super micron particles >5 μm, crystalline) and the mechanism of water uptake. The growth of droplet volume (Growth Factor, GF) for uric acid particles is experimentally observed for supermicrometer crystalline particles (>1 μm) at subsaturated humidity conditions (<100% RH). In addition, the water activity of submicrometer-size uric acid particles is estimated under subsaturated and supersaturated humidity conditions. These measurements provide us with information about the volume growth of droplets as water condenses in particles exposed to different humidity conditions. Our observations under subsaturated humidity conditions show that the uric acid particles have limited volume growth (<1% change per volume and <10% change per volume for crystalline and amorphous measurement, respectively). From the experimental data, the affinity of uric acid solute with water as a solvent is quantified in terms of water activity.

Water Transport-Induced Liquid-Liquid Phase Separation Facilitates Gelation for Controllable and Facile Fabrication of Physically Crosslinked Microgels.

Physically crosslinked microgels (PCMs) offer a biocompatible platform for various biomedical applications. However, current PCM fabrication methods suffer from their complexity and poor controllability, due to their reliance on altering physical conditions to initiate gelation and their dependence on specific materials. To address this issue, a novel PCM fabrication method is devised, which employs water transport-induced liquid-liquid phase separation (LLPS) to trigger the intermolecular interaction-supported sol-gel transition within aqueous emulsion droplets. This method enables the controllable and facile generation of PCMs through a single emulsification step, allowing for the facile production of PCMs with various materials and sizes, as well as controllable structures and mechanical properties. Moreover, this PCM fabrication method holds great promise for diverse biomedical applications. The interior of the PCM not only supports the encapsulation and proliferation of bacteria but also facilitates the encapsulation of eukaryotic cells after transforming the system into an all-aqueous emulsion. Furthermore, through appropriate surface functionalization, the PCMs effectively activate T cells in vitro upon coculturing. This work represents an advancement in PCM fabrication and offers new insights and perspectives for microgel engineering.

Spectral Characteristics of Unsaturated and Supersaturated Inorganic Aerosols: Insights into Deliquescence Kinetics.

The deliquescence phase transition of atmospheric aerosols is crucial for radiative forcing and atmospheric chemistry. However, the deliquescence kinetics of micrometer-sized aerosols, especially the formation and evolution of surface solution films, remain poorly understood. In this study, IR spectral characteristics were employed for the first time to quantify the solute concentration evolution in surface solution films. At a constant relative humidity (RH) of ∼65%, solution films on NaCl crystals exhibited a very low solute concentration (3.06 ± 0.18 mol/L), comparable to aqueous NaCl droplets above 90% RH. These films reached saturation at ∼74% RH, i.e., the deliquescence RH of NaCl, maintaining a nearly constant saturation level during deliquescence. In contrast, amorphous NaNO3 solids showed supersaturated solution films before deliquescence. Following deliquescence, the saturation level of solution phases increased due to faster solid dissolution rates than liquid water condensation. These findings address knowledge gaps in the complex nonequilibrium dissolution processes of crystalline or amorphous atmospheric aerosols.

Controlled Chemical‐Patterning of Textile to Accelerate Anti‐Gravity Water Flow

AbstractBio‐inspired unidirectional flow of tiny aqueous droplets across the fibrous substrate paved the way for the emergence of various advanced materials. In the past, textiles decorated with noncontact‐based wettability‐patterns enabled unidirectional water flow—without flooding the top surface by the transferred water. However, such approaches mostly suffer from a low (≈0.176 µL mm−2 s−1) flow rate and are likely to delay the overall liquid ejection process. Here, a chemically reactive coating capable of tailoring water wettability (121.3° ± 2.4° to 153.3° ± 1.8°) is introduced on commercially available textiles to develop chemically modulated wettability‐pattern for achieving a rapid (2.57 ± 0.28 µL mm−2 s−1) flow rate of water against the gravity with an ability to roll the accumulated liquids on the top surface. The spatially selected and controlled chemical modification with hydrophilic and hydrophobic small molecules through a 1, 4‐conjugate addition reaction yielded a 3D channel with a customized wettability gradient. The pinning and depinning of invaded water through such chemically decorated channels enabled unidirectional and fast penetration of liquid, where the water penetration resistance largely depends on the water penetration direction and dimension of the chemically modulated channels.

The effects of droplet stabilization by surfactants and nanoparticles on leakage, cross-talk, droplet stability, and cell adhesion.

Partially fluorinated nanoparticles (FNPs) have been proposed as a promising alternative for stabilising aqueous droplets in fluorinated oils. The exceptional energetic stability of FNPs at the droplet interface holds the potential for minimising leakage, enhancing stability, and promoting improved cell adhesion. However, their lower diffusion coefficient compared to surfactants presents challenges in achieving rapid droplet stabilisation, which is important in microfluidics applications. While several studies have focused on some of these aspects, a comprehensive study and direct comparison with conventional fluorosurfactants is still missing. In this manuscript, we undertake an examination and comparison of four crucial facets of both FNP- and surfactant-stabilised droplets: leakage of compounds, emulsion stability, droplet formation dynamics and cell adhesion. Contrary to what has previously been claimed, our findings demonstrate that FNPs only reduce leakage and cross-talk in very specific cases (e.g., resorufin), failing to provide enhanced compartmentalisation for highly hydrophobic dyes (e.g., rhodamine dyes). On the other hand, FNP-stabilised droplets indeed exhibit greater long-term stability compared to their surfactant-stabilised counterparts. Regarding the size of droplets generated via a diversity of microfluidic methods, no significant differences were observed between FNP-stabilised and surfactant-stabilised droplets. Finally, the previously reported improvements in cell adhesion and spreading on FNP-stabilised interfaces is limited to flat oil/water (o/w) interfaces and could not be observed within droplets. These comprehensive analyses shed light on the nuanced performance of FNPs and commercial fluorosurfactants as stabilising agents for aqueous droplets in fluorinated oils, contributing valuable insights for choosing the correct formulation for specific droplet-based microfluidics applications.

Development of surface droplet evaporation-based sensing platform for screening lipase inhibitors

The development of new types of sensing platforms with excellent performance is always appealing. In this report, we demonstrate a low-cost and user-friendly sensing platform based on drying of the aqueous droplet on the surface. The principle relies on the transduction of the surfactant concentration into the area of the dried droplet on the surface. The screening of a lipase inhibitor is demonstrated as an example. When triacylglycerols (GT) and lipase are mixed, the aqueous droplet produces a large dried area on the surface. This phenomenon is attributed to the production of sodium oleate due to the enzymatic digestion of GT by lipase. It results in reduction of the surface tension and causes wetting and spreading of the droplet, leaving a large dried area on the surface. However, in the presence of orlistat, a lipase inhibitor, a small dried area is obtained due to the inhibition of lipase activity by orlistat. Using this method, the lipase inhibitor can be readily and effectively screened. Compared to previous studies, this method is simple, rapid, low-cost and avoids the use of labelled molecules and complex instrumentation, with an IC50 value of 0.59 ± 0.09 μM for orlistat. In addition, it also works well with a portable microscope and a smartphone interface for data analysis. Overall, this study provides a new method for rapid and cost-effective screening of lipase inhibitors.

1-Octanol-assisted ultra-small volume droplet microfluidics with nanoelectrospray ionization mass spectrometry

BackgroundDroplet microfluidics with push-pull and microdialysis sampling from brain slices, cultured cells and engineered tissues produce low volume mass limited samples containing analytes sampled from the extracellular space. This sampling approach coupled to mass spectrometry (MS) detection allows evaluation of time-dependent chemical changes. Our goal is an approach for continuous sampling and segregation of extracellular samples into picoliter droplets followed by the characterization of the droplets using nanoelectrospray ionization (nESI) MS. The main focus here is the optimization of the carrier oil for the microfluidic device that neither affects the stability of picoliter droplets nor compatibility with MS detection of a range of analytes. ResultsWe developed and characterized a 1-octanol-assisted ultra-small volume droplet microfluidic nESI MS system for the analysis of neurotransmitters in distinct samples including cerebrospinal fluid (CSF). The use of a 1-octanol oil phase was effective for generation of aqueous droplets as small as 65 pL and enabled detection of acetylcholine (ACh) and gamma-aminobutyric acid (GABA) in water and artificial CSF. Continuous MS analysis of droplets for extended periods up to 220 min validated the long-term stability of droplet generation and analyte detection by nESI-MS. As an example, ACh response demonstrated a linear working range (R2 = 0.99) between 0.4 μM and 25 μM with a limit of detection of 370 nM (24 amol), enabling its quantitation in rodent CSF. SignificanceThe established droplet microfluidics – nESI MS approach allows the analysis of microenvironments at high spatiotemporal resolution. The approach may allow microsampling and monitoring of spatiotemporal dynamics of neurochemicals and drugs in the brain and spinal cord of live animals.

Transparency-changing elastomers by controlling of the refractive index of liquid inclusions

Complex materials that change their optical properties in response to changes in environmental conditions can find applications in displays, smart windows, and optical sensors. Here a class of biphasic composites with stimuli-adaptive optical transmittance is introduced. The biphasic composites comprise aqueous droplets (a mixture of water, glycerol, and surfactant) embedded in an elastomeric matrix. The biphasic composites are tuned to be optically transparent through a careful match of the refractive indices between the aqueous droplets and the elastomeric matrix. We demonstrate that stimuli (e.g., salinity and temperature change) can trigger variations in the optical transmittance of the biphasic composite. The introduction of such transparency-changing soft matter with liquid inclusions offers a novel approach to designing advanced optical devices, optical sensors, and metamaterials.