Two-dimensional perfusion angiography permits direct visualization of redistribution of flow in hepatocellular carcinoma during b-TACE.

OBJECTIVES: To demonstrate in vivo redistribution of the blood flow towards HCC's lesions by utilizing two-dimensional perfusion angiography in b-TACE procedures. MATERIAL AND METHODS: In total, 30 patients with 35 HCC nodules treated in the period between January 2019 and November 2021. For each patient, a post-processing software leading to a two-dimensional perfusion angiography was applied on each angiography performed via balloon microcatheter, before and after inflation. On the colour map obtained, reflecting the evolution of contrast intensity change over time, five regions of interests (ROIs) were assessed: one on the tumour (ROI-t), two in the immediate peritumoural healthy liver parenchyma (ROI-ihl) and two in the peripheral healthy liver parenchyma (ROI-phl). The results have been interpreted with a novel in silico model that simulates the hemodynamics of the hepatic arterial system. RESULTS: Among the ROIs drawn inside the same segment of target lesion, the time-to-peak of the ROI-t and of the ROI-ihl have a significantly higher mean value when the balloon was inflated compared with the ROIs obtained with deflated balloon (10.33 ± 3.66 s vs 8.87 ± 2.60 s (p = 0.015) for ROI-t; 10.50 ± 3.65 s vs 9.23 ± 2.70 s (p = 0.047) for ROI-ihl). The in silico model prediction time-to-peak delays when balloon was inflated, match with those observed in vivo. The numerical flow analysis shows how time-to-peak delays are caused by the obstruction of the balloon-occluded artery and the opening of intra-hepatic collateral. CONCLUSION: The measurements identify predictively the flow redistribution in the hepatic arteries during b-TACE, supporting a proper positioning of the balloon microcatheter.

Microfluidic Fabrication of Monodisperse and Recyclable TiO2-Poly(ethylene glycol) Diacrylate Hybrid Microgels for Removal of Methylene Blue from Aqueous Medium.

Nearly monodisperse titanium oxide-polyethylene glycol diacrylate [TiO2-P(EGDA)] hybrid microbeads containing 0.5 wt % TiO2 nanoparticles entrapped within a P(EGDA) cross-linked polymeric network were synthesized using a modular Lego-inspired glass capillary microfluidic device. TiO2-P(EGDA) hybrid microgels were characterized by optical microscopy, scanning electron microscopy, X-ray diffraction, energy dispersive X-ray spectroscopy, and thermogravimetric analysis. The fabricated TiO2-P(EGDA) hybrid microgel system showed 100% removal efficiency of methylene blue (MB) from its 1-3 ppm aqueous solutions after 4 h of UV light irradiation at 0.2 mW/cm2 at the loading of 25 g/L photocatalyst beads in the reaction mixture, corresponding to the loading of naked TiO2 of just 0.025 g/L. No decrease in photocatalytic efficiency was observed in 10 repeated runs with recycled photocatalyst using a fresh 1 ppm MB solution in each cycle. The rate of photocatalytic degradation was controlled by the UV light irradiance, catalyst loading, and the initial dye concentration. Physical adsorption of MB onto the surface of composite microgel was also observed. The adsorption data was best fitted with the Langmuir adsorption isotherm and the Elovich kinetic model. TiO2-P(EGDA) microgel beads are biocompatible, can be prepared with a tunable size in the microfluidic device, and can easily be separated from the reaction mixture by gravity settling. The TiO2-P(EGDA) system can be used for the removal of other toxic dyes and micropollutants from industrial wastewater.

Enhanced Accumulation of Colloidal Particles in Microgrooved Channels via Diffusiophoresis and Steady-State Electrolyte Flows.

The delivery of colloidal particles in dead-end microstructures is very challenging, since these geometries do not allow net flows of particle-laden fluids; meanwhile, diffusive transport is slow and inefficient. Recently, we introduced a novel particle manipulation strategy, based on diffusiophoresis, whereby the salt concentration gradient between parallel electrolyte streams in a microgrooved channel induces the rapid (i.e., within minutes) and reversible accumulation, retention, and removal of colloidal particles in the microgrooves. In this study, we investigated the effects of salt contrast and groove depth on the accumulation process in silicon microgrooves and determined the experimental conditions that lead to a particle concentration peak of more than four times the concentration in the channel bulk. Also, we achieved an average particle concentration in the grooves of more than twice the concentration in the flowing streams and almost 2 orders of magnitude larger than the average concentration in the grooves in the absence of a salt concentration gradient. Analytical sufficient and necessary conditions for particle accumulation are also derived. Finally, we successfully tested the accumulation process in polydimethylsiloxane microgrooved channels, as they are less expensive to fabricate than silicon microgrooved substrates. The controlled and enhanced accumulation of colloidal particles in dead-end structures by solute concentration gradients has potential applications in soft matter and living systems, such as drug delivery, synthetic biology, and on-chip diagnostics.

Facile Microfluidic Fabrication of Biocompatible Hydrogel Microspheres in a Novel Microfluidic Device.

Poly(ethylene glycol) diacrylate (PEGDA) microgels with tuneable size and porosity find applications as extracellular matrix mimics for tissue-engineering scaffolds, biosensors, and drug carriers. Monodispersed PEGDA microgels were produced by modular droplet microfluidics using the dispersed phase with 49-99 wt% PEGDA, 1 wt% Darocur 2959, and 0-50 wt% water, while the continuous phase was 3.5 wt% silicone-based surfactant dissolved in silicone oil. Pure PEGDA droplets were fully cured within 60 s at the UV light intensity of 75 mW/cm2. The droplets with higher water content required more time for curing. Due to oxygen inhibition, the polymerisation started in the droplet centre and advanced towards the edge, leading to a temporary solid core/liquid shell morphology, confirmed by tracking the Brownian motion of fluorescent latex nanoparticles within a droplet. A volumetric shrinkage during polymerisation was 1-4% for pure PEGDA droplets and 20-32% for the droplets containing 10-40 wt% water. The particle volume increased by 36-50% after swelling in deionised water. The surface smoothness and sphericity of the particles decreased with increasing water content in the dispersed phase. The porosity of swollen particles was controlled from 29.7% to 41.6% by changing the water content in the dispersed phase from 10 wt% to 40 wt%.

Crosslinking Strategies for the Microfluidic Production of Microgels.

This article provides a systematic review of the crosslinking strategies used to produce microgel particles in microfluidic chips. Various ionic crosslinking methods for the gelation of charged polymers are discussed, including external gelation via crosslinkers dissolved or dispersed in the oil phase; internal gelation methods using crosslinkers added to the dispersed phase in their non-active forms, such as chelating agents, photo-acid generators, sparingly soluble or slowly hydrolyzing compounds, and methods involving competitive ligand exchange; rapid mixing of polymer and crosslinking streams; and merging polymer and crosslinker droplets. Covalent crosslinking methods using enzymatic oxidation of modified biopolymers, photo-polymerization of crosslinkable monomers or polymers, and thiol-ene "click" reactions are also discussed, as well as methods based on the sol-gel transitions of stimuli responsive polymers triggered by pH or temperature change. In addition to homogeneous microgel particles, the production of structurally heterogeneous particles such as composite hydrogel particles entrapping droplet interface bilayers, core-shell particles, organoids, and Janus particles are also discussed. Microfluidics offers the ability to precisely tune the chemical composition, size, shape, surface morphology, and internal structure of microgels by bringing multiple fluid streams in contact in a highly controlled fashion using versatile channel geometries and flow configurations, and allowing for controlled crosslinking.

Reversible Trapping of Colloids in Microgrooved Channels via Diffusiophoresis under Steady-State Solute Gradients.

The controlled transport of colloids in dead-end structures is a key capability that can enable a wide range of applications, such as biochemical analysis, drug delivery, and underground oil recovery. This Letter presents a new trapping mechanism that allows the fast (i.e., within a few minutes) and reversible accumulation of submicron particles within dead-end microgrooves by means of parallel streams with different salinity level. For the first time, particle focusing in dead-end structures is achieved under steady-state gradients. Confocal microscopy analysis and numerical investigations show that the particles are trapped at a flow recirculation region within the grooves due to a combination of diffusiophoresis transport and hydrodynamic effects. Counterintuitively, the particle velocity at the focusing point is not vanishing and, hence, the particles are continuously transported in and out of the focusing point. The accumulation process is also reversible and one can cyclically trap and release the colloids by controlling the salt concentration of the streams via a flow switching valve.

Highly Porous Magnetic Janus Microparticles with Asymmetric Surface Topology.

Monodispersed magnetic Janus particles composed of a porous polystyrene portion and a nonporous poly(vinyl acetate) portion with embedded oleic acid-coated magnetic nanoparticles were generated using microfluidic emulsification followed by two distinct phase separation events triggered by solvent evaporation. The template droplets were composed of 2 wt % polystyrene, 2 wt % poly(vinyl acetate), and 0.5-2 wt % n-heptane-based magnetic fluid dissolved in dichloromethane (DCM). The porosity of polystyrene compartments was the result of phase separation between a nonvolatile nonsolvent (n-heptane) and a volatile solvent (DCM) within polystyrene-rich phase. The focused ion beam cross-sectioning and scanning electron microscopy (SEM) imaging revealed high surface porosity of polystyrene compartments with negligible porosity of poly(vinyl acetate) parts, which can be exploited to increase the wettability contrast between the two polymers and enhance bubble generation in bubble-driven micromotors. The porosity of the polystyrene portion was controlled by varying the fraction of n-heptane in the dispersed phase. The particle composition was confirmed by scanning electron microscopy-energy-dispersive X-ray spectroscopy, Fourier transform infrared spectroscopy, and differential scanning calorimetry. The fabricated particles were successfully magnetized when subjected to an external magnetic field, which led to their aggregation into regular 2D assemblies. The particle clusters composed of two to four individual particles could be rotated with a rotating magnetic field. Microfluidic generation of highly porous Janus particles with compositional, topological, and magnetic asymmetry provides a cost-effective, easy-to-implement yet highly robust and versatile strategy for the manufacturing of multifunctional smart particles.

Mask-Free Laser Lithography for Rapid and Low-Cost Microfluidic Device Fabrication.

Microfluidics has become recognized as a powerful platform technology associated with a constantly increasing array of applications across the life sciences. This surge of interest over recent years has led to an increased demand for microfluidic chips, resulting in more time being spent in the cleanroom fabricating devices using soft lithography-a slow and expensive process that requires extensive materials, training and significant engineering resources. This bottleneck limits platform complexity as a byproduct of lengthy delays between device iterations and affects the time spent developing the final application. To address this problem, we report a new, rapid, and economical approach to microfluidic device fabrication using dry resist films to laminate laser cut sheets of acrylic. We term our method laser lithography and show that our technique can be used to engineer 200 µm width channels for assembling droplet generators capable of generating monodisperse water droplets in oil and micromixers designed to sustain chemical reactions. Our devices offer high transparency, negligible device to device variation, and low X-ray background scattering, demonstrating their suitability for real-time X-ray-based characterization applications. Our approach also requires minimal materials and apparatus, is cleanroom free, and at a cost of around $1.00 per chip could significantly democratize device fabrication, thereby increasing the interdisciplinary accessibility of microfluidics.

Microfluidic Production of Poly(1,6-hexanediol diacrylate)-Based Polymer Microspheres and Bifunctional Microcapsules with Embedded TiO2 Nanoparticles.

Application of TiO2 as a photocatalyst and UV protector is restricted by the difficulties in the recovery of TiO2 nanoparticles after water treatment. In this work, TiO2 nanoparticles (Degussa P25) were immobilised within easily recoverable poly(1,6-hexanediol diacrylate) poly(HDDA)-based polymer microspheres produced by on-the-fly photopolymerization of microfluidically generated droplets. Because of fast polymerization reaction, TiO2 was uniformly distributed within the polymer network. The transformation of double bonds in terminal vinyl groups of HDDA monomer into single bonds during photopolymerization was confirmed by Fourier transform infrared spectroscopy. The microspheres containing 0.5 wt % TiO2 embedded in a poly(HDDA) matrix degraded 80% of methylene blue from 1 ppm aqueous solution in 9 h under UV light irradiation of 0.9 mW/cm2 at 365 nm. The microspheres could easily be separated from water and used in repeated cycles without any loss in photocatalytic activity. The inclusion of TiO2 within a polymer matrix increased the thermal degradation temperature of the material from 364 to 389 °C. Bifunctional microcapsules consisting of aqueous or liquid paraffin core enclosed within a TiO2/poly(HDDA) composite polymer shell were also prepared. The fluorescent dye calcein was encapsulated in the core with 100% efficiency.

Mechanical Characterization of Ultralow Interfacial Tension Oil-in-Water Droplets by Thermal Capillary Wave Analysis in a Microfluidic Device.

Measurements of the ultralow interfacial tension and surfactant film bending rigidity for micron-sized heptane droplets in bis(2-ethylhexyl) sodium sulfosuccinate-NaCl aqueous solutions were performed in a microfluidic device through the analysis of thermally driven droplet interface fluctuations. The Fourier spectrum of the stochastic droplet interface displacement was measured through bright-field video microscopy and a contour analysis technique. The droplet interfacial tension, together with the surfactant film bending rigidity, was obtained by fitting the experimental results to the prediction of a capillary wave model. Compared to existing methods for ultralow interfacial tension measurements, this contactless, nondestructive, all-optical approach has several advantages, such as fast measurement, easy implementation, cost-effectiveness, reduced amount of liquids, and integration into lab-on-a-chip devices.

Optically assembled droplet interface bilayer (OptiDIB) networks from cell-sized microdroplets.

We report a new platform technology to systematically assemble droplet interface bilayer (DIB) networks in user-defined 3D architectures from cell-sized droplets using optical tweezers. Our OptiDIB platform is the first demonstration of optical trapping to precisely construct 3D DIB networks, paving the way for the development of a new generation of modular bio-systems.

Selective Adsorption of Ionic Species Using Macroporous Monodispersed Polyethylene Glycol Diacrylate/Acrylic Acid Microgels with Tunable Negative Charge.

Monodispersed polyethylene glycol diacrylate (PEGDA)/acrylic acid (AA) microgels with a tuneable negative charge and macroporous internal structure have been produced using a Lego-inspired droplet microfluidic device. The surface charge of microgels was controlled by changing the content of AA in the monomer mixture from zero (for noncharged PEGDA beads) to 4 wt%. The macroporosity of the polymer matrix was introduced by adding 20 wt% of 600-MW polyethylene glycol (PEG) as a porogen material into the monomer mixture. The porogen was successfully leached out with acetone after UV-crosslinking, which resulted in micron-sized cylindrical pores with crater-like morphology, uniformly arranged on the microgel surface. Negatively charged PEGDA/AA beads showed improved adsorption capacity towards positively charged organic dyes (methylene blue and rhodamine B) compared to neutral PEGDA beads and high repulsion of negatively charged dye molecules (methyl orange and congo red). Macroporous microgels showed better adsorption properties than nonporous beads, with a maximum adsorption capacity towards methylene blue of 45 mg/g for macroporous PEGDA/AA microgels at pH 8.6, as compared to 23 mg/g for nonporous PEGDA/AA microgels at the same pH. More than 98% of Cu(II) ions were removed from 50 ppm solution at pH 6.7 using 2.7 mg/mL of macroporous PEGDA/AA microgel. The adsorption of cationic species was significantly improved when pH was increased from 3 to 9 due to a higher degree of ionization of AA monomeric units in the polymer network. The synthesized copolymer beads can be used in drug delivery to achieve improved loading capacity of positively charged therapeutic agents and in tissue engineering, where a negative charge of scaffolds coupled with porous structure can help to achieve improved permeability of high-molecular-weight metabolites and nutrients, and anti-fouling activity against negatively charged species.

Continuous Manipulation and Characterization of Colloidal Beads and Liposomes via Diffusiophoresis in Single- and Double-Junction Microchannels.

We reveal a physical mechanism that enables the preconcentration, sorting, and characterization of charged polystyrene nanobeads and liposomes dispersed in a continuous flow within a straight micron-sized channel. Initially, a single Ψ-junction microfluidic chip is used to generate a steady-state salt concentration gradient in the direction perpendicular to the flow. As a result, fluorescent nanobeads dispersed in the electrolyte solutions accumulate into symmetric regions of the channel, appearing as two distinct symmetric stripes when the channel is observed from the top via epi-fluorescence microscopy. Depending on the electrolyte flow configuration and, thus, the direction of the salt concentration gradient field, the fluorescent stripes get closer to or apart from each other as the distance from the inlet increases. Our numerical and experimental analysis shows that although nanoparticle diffusiophoresis and hydrodynamic effects are involved in the accumulation process, diffusio-osmosis along the top and bottom channel walls plays a crucial role in the observed particles dynamics. In addition, we developed a proof-of-concept double Ψ-junction microfluidic device that exploits this accumulation mechanism for the size-based separation and size detection of nanobeads as well as for the measurement of zeta potential and charged lipid composition of liposomes under continuous flow settings. This device is also used to investigate the effect of fluid-like or gel-like states of the lipid membranes on the liposome diffusiophoretic response. The proposed strategy for solute-driven manipulation and characterization of colloids has great potential for microfluidic bioanalytical testing applications, including bioparticle preconcentration, sorting, sensing, and analysis.

Lego-Inspired Glass Capillary Microfluidic Device: A Technique for Bespoke Microencapsulation of Phase Change Materials.

We report a Lego-inspired glass capillary microfluidic device capable of encapsulating both organic and aqueous phase change materials (PCMs) with high reproducibility and 100% PCM yield. Oil-in-oil-in-water (O/O/W) and water-in-oil-in-water (W/O/W) core-shell double emulsion droplets were formed to encapsulate hexadecane (HD, an organic PCM) and salt hydrate SP21EK (an aqueous PCM) in a UV-curable polymeric shell, Norland Optical Adhesive (NOA). The double emulsions were consolidated through on-the-fly polymerization, which followed thiol-ene click chemistry for photoinitiation. The particle diameters and shell thicknesses of the microcapsules were controlled by manipulating the geometry of glass capillaries and fluid flow rates. The microcapsules were monodispersed and exhibited the highest encapsulation efficiencies of 65.4 and 44.3% for HD and SP21EK-based materials, respectively, as determined using differential scanning calorimetry (DSC). The thermogravimetric (TGA) analysis confirmed much higher thermal stability of both encapsulated PCMs compared to pure PCMs. Polarization microscopy revealed that microcapsules could sustain over 100 melting-crystallization cycles without any structural changes. Bifunctional microcapsules with remarkable photocatalytic activity along with thermal energy storage performance were produced after the addition of 1 wt % titanium dioxide (TiO2) nanoparticles (NPs) into the polymeric shell. The presence of TiO2 NPs in the shell was confirmed by higher opacity and whiteness of these microcapsules and was quantified by energy dispersive X-ray (EDX) spectroscopy. Young's modulus of HD-based microcapsules estimated using micromanipulation analysis increased from 58.5 to 224 MPa after TiO2 incorporation in the shell.

Monodispersed Sirolimus-Loaded PLGA Microspheres with a Controlled Degree of Drug-Polymer Phase Separation for Drug-Coated Implantable Medical Devices and Subcutaneous Injection.

Monodispersed sirolimus (SRL)-loaded poly(lactic-co-glycolic acid) microspheres with a diameter of 1.8, 3.8, and 8.5 µm were produced by high-throughput microfluidic step emulsification─solvent evaporation using single crystal silicon chips consisted of 540-1710 terraced microchannels with a depth of 2, 4, or 5 µm arranged in 10 parallel arrays. Uniform sized droplets were generated over 25 h across all channels. Nearly 15% of the total drug was released by the initial burst release during an accelerated drug release testing performed at 37 °C using a hydrotropic solution containing 5.8 M N,N-diethylnicotinamide. After 24 h, 71% of the drug was still entrapped in the particles. The internal morphology of microspheres was investigated by fluorescence microscopy using Nile red as a selective fluorescent stain with higher binding affinity toward SRL. By increasing the drug loading from 33 to 50 wt %, the particle morphology evolved from homogeneous microspheres, in which the drug and polymer were perfectly mixed, to patchy particles, with amorphous drug patches embedded within a polymer matrix to anisotropic patchy Janus particles. Janus particles with fully segregated drug and polymer regions were achieved by pre-saturating the aqueous phase with the organic solvent, which decreased the rate of solvent evaporation and allowed enough time for complete phase separation. This approach to manufacturing drug-loaded monodisperse microparticles can enable the development of more effective implantable drug-delivery devices and improved methods for subcutaneous drug administration, which can lead to better therapeutic treatments.

Engineering motile aqueous phase-separated droplets via liposome stabilisation.

There are increasing efforts to engineer functional compartments that mimic cellular behaviours from the bottom-up. One behaviour that is receiving particular attention is motility, due to its biotechnological potential and ubiquity in living systems. Many existing platforms make use of the Marangoni effect to achieve motion in water/oil (w/o) droplet systems. However, most of these systems are unsuitable for biological applications due to biocompatibility issues caused by the presence of oil phases. Here we report a biocompatible all aqueous (w/w) PEG/dextran Pickering-like emulsion system consisting of liposome-stabilised cell-sized droplets, where the stability can be easily tuned by adjusting liposome composition and concentration. We demonstrate that the compartments are capable of negative chemotaxis: these droplets can respond to a PEG/dextran polymer gradient through directional motion down to the gradient. The biocompatibility, motility and partitioning abilities of this droplet system offers new directions to pursue research in motion-related biological processes.

Fusing Artificial Cell Compartments and Lipid Domains Using Optical Traps: A Tool to Modulate Membrane Composition and Phase Behaviour.

New technologies for manipulating biomembranes have vast potential to aid the understanding of biological phenomena, and as tools to sculpt novel artificial cell architectures for synthetic biology. The manipulation and fusion of vesicles using optical traps is amongst the most promising due to the level of spatiotemporal control it affords. Herein, we conduct a suite of feasibility studies to show the potential of optical trapping technologies to (i) modulate the lipid composition of a vesicle by delivering new membrane material through fusion events and (ii) manipulate and controllably fuse coexisting membrane domains for the first time. We also outline some noteworthy morphologies and transitions that the vesicle undergoes during fusion, which gives us insight into the mechanisms at play. These results will guide future exploitation of laser-assisted membrane manipulation methods and feed into a technology roadmap for this emerging technology.

3D Arrays of Super-Hydrophobic Microtubes from Polypore Mushrooms as Naturally-Derived Systems for Oil Absorption.

Porous materials derived from natural resources, such as Luffa sponges, pomelo peel and jute fibres, have recently emerged as oil adsorbents for water purification, due to their suitability, low environmental impact, biodegradability and low cost. Here we show, for the first time, that the porosity of the fruiting body of polypore mushrooms can be used to absorb oils and organic solvents while repelling water. We engineered the surface properties of Ganoderma applanatum fungi, of which the fruiting body consists of a regular array of long capillaries embedded in a fibrous matrix, with paraffin wax, octadecyltrichlorosilane (OTS) and trichloro(1H,1H,2H,2H-perfluorooctyl)silane. Morphological and wettability analyses of the modified fungus revealed that the OTS treatment was effective in preserving the 3D porosity of the natural material, inducing super-hydrophobicity (water contact angle higher than 150°) and improving oil sorption capacity (1.8⁻3.1 g/g). The treated fungus was also inserted into fluidic networks as a filtration element, and its ability to separate water from chloroform was demonstrated.

Sculpting and fusing biomimetic vesicle networks using optical tweezers.

Constructing higher-order vesicle assemblies has discipline-spanning potential from responsive soft-matter materials to artificial cell networks in synthetic biology. This potential is ultimately derived from the ability to compartmentalise and order chemical species in space. To unlock such applications, spatial organisation of vesicles in relation to one another must be controlled, and techniques to deliver cargo to compartments developed. Herein, we use optical tweezers to assemble, reconfigure and dismantle networks of cell-sized vesicles that, in different experimental scenarios, we engineer to exhibit several interesting properties. Vesicles are connected through double-bilayer junctions formed via electrostatically controlled adhesion. Chemically distinct vesicles are linked across length scales, from several nanometres to hundreds of micrometres, by axon-like tethers. In the former regime, patterning membranes with proteins and nanoparticles facilitates material exchange between compartments and enables laser-triggered vesicle merging. This allows us to mix and dilute content, and to initiate protein expression by delivering biomolecular reaction components.

Engineering thermoresponsive phase separated vesicles formed via emulsion phase transfer as a content-release platform.

Giant unilamellar vesicles (GUVs) are a well-established tool for the study of membrane biophysics and are increasingly used as artificial cell models and functional units in biotechnology. This trend is driven by the development of emulsion-based generation methods such as Emulsion Phase Transfer (EPT), which facilitates the encapsulation of almost any water-soluble compounds (including biomolecules) regardless of size or charge, is compatible with droplet microfluidics, and allows GUVs with asymmetric bilayers to be assembled. However, the ability to control the composition of membranes formed via EPT remains an open question; this is key as composition gives rise to an array of biophysical phenomena which can be used to add functionality to membranes. Here, we evaluate the use of GUVs constructed via this method as a platform for phase behaviour studies and take advantage of composition-dependent features to engineer thermally-responsive GUVs. For the first time, we generate ternary GUVs (DOPC/DPPC/cholesterol) using EPT, and by compensating for the lower cholesterol incorporation efficiencies, show that these possess the full range of phase behaviour displayed by electroformed GUVs. As a demonstration of the fine control afforded by this approach, we demonstrate release of dye and peptide cargo when ternary GUVs are heated through the immiscibility transition temperature, and show that release temperature can be tuned by changing vesicle composition. We show that GUVs can be individually addressed and release triggered using a laser beam. Our findings validate EPT as a suitable method for generating phase separated vesicles and provide a valuable proof-of-concept for engineering content release functionality into individually addressable vesicles, which could have a host of applications in the development of smart synthetic biosystems.