Effects Of Liquid-liquid Phase Separation Research Articles

New insights into pure zwitterionic hydrogels with high strength and high toughness.

Zwitterionic hydrogels are electrically neutral materials with both cationic and anionic groups that impart excellent anti-fouling properties and ion channel orientations. However, pure zwitterionic hydrogels generally exhibit low strength and toughness. In this study, it has been discovered that polymerizable zwitterionic monomers in aqueous solution exhibit a unique liquid-liquid phase separation phenomenon at a high monomer concentration of ≥50 wt%, resulting in pure and commercial zwitterionic hydrogels with high compressive strength (6.5 MPa) and high toughness (2.12 kJ m-2). This phase separation and the corresponding aggregations might be caused by strong dipole-dipole interactions among residual zwitterionic monomers under the lack of free-water condition. The synergistic effect of liquid-liquid phase separation and polymer entanglement enhances the mechanical strength, toughness, self-recovery, and anti-freezing properties of pure polyzwitterionic hydrogels. Moreover, the high fracture energy of highly elongated yet tough polyzwitterionic hydrogels facilitates the development of high crack propagation resistance, which supports an expanded role in tissue engineering, soft flexible devices, and electronics applications with improved durability. A wide range of applications for the proposed polyzwitterionic hydrogels is demonstrated by the development and testing of a strain sensor and a triboelectric nanogenerator device. Our findings provide novel insights into the network structure of pure polyzwitterionic hydrogels.

Self-assembly coupled to liquid-liquid phase separation.

Liquid condensate droplets with distinct compositions of proteins and nucleic acids are widespread in biological cells. While it is known that such droplets, or compartments, can regulate irreversible protein aggregation, their effect on reversible self-assembly remains largely unexplored. In this article, we use kinetic theory and solution thermodynamics to investigate the effect of liquid-liquid phase separation on the reversible self-assembly of structures with well-defined sizes and architectures. We find that, when assembling subunits preferentially partition into liquid compartments, robustness against kinetic traps and maximum achievable assembly rates can be significantly increased. In particular, both the range of solution conditions leading to productive assembly and the corresponding assembly rates can increase by orders of magnitude. We analyze the rate equation predictions using simple scaling estimates to identify effects of liquid-liquid phase separation as a function of relevant control parameters. These results may elucidate self-assembly processes that underlie normal cellular functions or pathogenesis, and suggest strategies for designing efficient bottom-up assembly for nanomaterials applications.

Water uptake of subpollen aerosol particles: hygroscopic growth, cloud condensation nuclei activation, and liquid–liquid phase separation

Abstract. Pollen grains emitted from vegetation can release subpollen particles (SPPs) that contribute to the fine fraction of atmospheric aerosols and may act as cloud condensation nuclei (CCN), ice nuclei (IN), or aeroallergens. Here, we investigate and characterize the hygroscopic growth and CCN activation of birch, pine, and rapeseed SPPs. A high-humidity tandem differential mobility analyzer (HHTDMA) was used to measure particle restructuring and water uptake over a wide range of relative humidity (RH) from 2 % to 99.5 %, and a continuous flow CCN counter was used for size-resolved measurements of CCN activation at supersaturations (S) in the range of 0.2 % to 1.2 %. For both subsaturated and supersaturated conditions, effective hygroscopicity parameters, κ, were obtained by Köhler model calculations. Gravimetric and chemical analyses, electron microscopy, and dynamic light scattering measurements were performed to characterize further properties of SPPs from aqueous pollen extracts such as chemical composition (starch, proteins, DNA, and inorganic ions) and the hydrodynamic size distribution of water-insoluble material. All investigated SPP samples exhibited a sharp increase of water uptake and κ above ∼95 % RH, suggesting a liquid–liquid phase separation (LLPS). The HHTDMA measurements at RH >95 % enable closure between the CCN activation at water vapor supersaturation and hygroscopic growth at subsaturated conditions, which is often not achieved when hygroscopicity tandem differential mobility analyzer (HTDMA) measurements are performed at lower RH where the water uptake and effective hygroscopicity may be limited by the effects of LLPS. Such effects may be important not only for closure between hygroscopic growth and CCN activation but also for the chemical reactivity, allergenic potential, and related health effects of SPPs.

Alteration of Protein Binding Affinities by Aqueous Two-Phase Systems Revealed by Pressure Perturbation

Interactions between proteins and ligands, which are fundamental to many biochemical processes essential to life, are mostly studied at dilute buffer conditions. The effects of the highly crowded nature of biological cells and the effects of liquid-liquid phase separation inducing biomolecular droplet formation as a means of membrane-less compartmentalization have been largely neglected in protein binding studies. We investigated the binding of a small ligand (ANS) to one of the most multifunctional proteins, bovine serum albumin (BSA) in an aqueous two-phase system (ATPS) composed of PEG and Dextran. Also, aiming to shed more light on differences in binding mode compared to the neat buffer data, we examined the effect of high hydrostatic pressure (HHP) on the binding process. We observe a marked effect of the ATPS on the binding characteristics of BSA. Not only the binding constants change in the ATPS system, but also the integrity of binding sites is partially lost, which is most likely due to soft enthalpic interactions of the BSA with components in the dense droplet phase of the ATPS. Using pressure modulation, differences in binding sites could be unravelled by their different volumetric and hydration properties. Regarding the vital biological relevance of the study, we notice that extreme biological environments, such as HHP, can markedly affect the binding characteristics of proteins. Hence, organisms experiencing high-pressure stress in the deep sea need to finely adjust the volume changes of their biochemical reactions in cellulo.

Effect of lower critical solution temperature phase separation on crystallization kinetics and morphology of poly(butylenes succinate)/poly(ethylene oxide) blend

The crystalline/crystalline polymer blend of poly(butylenes succinate)/poly(ethylene oxide) (PBS/PEO) exhibited lower critical solution temperature (LCST) phase separation behavior at temperatures above the melting point of PBS, which was measured by phase contrast optical microscopy (PCOM) technique. In this work, we determined the LCST phase diagram of PBS/PEO blends, and then investigated the effect of liquid–liquid phase separation (LLPS) on the crystallization kinetics and morphologies of PBS/PEO (40/60 wt/wt) blend. Based on polarized optical microscopy (POM), scanning electron microscopy (SEM) and differential scanning calorimeter (DSC) results, we found that the spherulite morphology and crystallization kinetics of PBS were strongly influenced by the initial LLPS structure; because the kinetics of liquid–liquid phase dissolution was much slower than that of crystallization. The effect of LLPS on the primary nucleation density and crystal growth rate of PBS were strongly dependent on the crystallization temperature of PBS. After a second quench to a temperature below the melting point of PEO, the fractionated crystallization of PEO was observed. The competition between the kinetics of LLPS and the driving force of PBS crystallization controlled the spatial distribution of PEO in PBS spherulites, thus essentially determined the confined conditions of the PEO crystallization. The results indicated that the LLPS was an effective process to control the crystallization kinetics and morphology in crystalline/crystalline polymer blend systems.

Liquid-Liquid Phase Separation in Photo-Thermo-Refractive Glass

Photo-thermo-refractive (PTR) glass is an optical Na–Al–Zn–K–O–F–Br silicate glass doped with Ag, Ce, Sb, and Sn that undergoes photo-thermo-induced volume crystallization of nanosized NaF responsible for localized refractive index changes. PTR glass has found numerous commercial applications, but the intricate mechanism of photo-thermo crystallization is far from being understood. In this paper, we demonstrate that, additional to crystalline phase precipitation, liquid–liquid phase separation (LLPS), i.e. amorphous droplets embedded in the matrix glass, appears concurrently over a wide range of temperatures. The immiscibility temperature is 925°C. The droplet phase is richer in SiO2, rendering the alkali-rich remaining matrix glass a lower glass transition temperature and a higher electrical conductivity than the original glass. The droplet's surface does not catalyze NaF nucleation. Although the effects of LLPS on optical properties of PTR glass are still to be explored, it could contribute to unwanted scattering losses and/or uncontrolled refractive index change. The substantial change in the original glass composition resulting from LLPS should play an important role on NaF crystallization kinetics, and therefore must be considered for an overall understanding of the crystallization mechanism underpinning the refractive index change in PTR glass.

Effects of spinodal decomposition on mechanical properties of a polyolefin blend from high to low strain rates

The effects of spinodal decomposition, a typical type of liquid–liquid phase separation (LLPS), on the mechanical properties of a pretreated statistical copolymer blend of poly(ethylene- co-hexene) (PEH) and poly(ethylene- co-butene) (PEB) were characterized by tensile testing under different strain rates. An important finding was that the strain rate and the crystallization temperature had to be considered as independent variables in analyzing the effects of spinodal decomposition on the tensile behaviors. At the high strain rate, the stress–strain curves kept irrespective of LLPS time, in which the interfacial relaxation between phase domains could not be detected, except the case crystallizing at 120 °C for 10 min. This was explained in terms of the distribution of the crystals elaborated by differential scanning calorimetry (DSC) results. However, when a relatively low strain rate was employed, a clear deterioration of tensile properties with LLPS proceeding was observed for the cases with low crystallization temperature because of its detection ability for large scale structural information, such as the phase boundary; unexpectedly, the effect of LLPS on the tensile properties was found to disappear in the high crystallization temperature cases which was due to the cooperative functions of the phase boundary and the internal structures of the phase domains. These abundant results provided a novel and indispensable instruction for the processing of polymer blends from the theoretical viewpoint.

Dielectric relaxation and crystallization of nanophase separated 1-propanol-isoamylbromide mixture

The effects of liquid-liquid phase separation on molecular relaxation of an apparently homogeneous mixture of 1-propanol and isoamylbromide has been studied by dielectric spectroscopy over a broad frequency and temperature range, and its crystallization kinetics investigated in real time. The mixture shows two widely separated relaxation processes, as before, with the faster relaxation due to the orientational diffusion of isoamylbromide and the slower due to that of 1-propanol. In the mixture, the scaled contribution to permittivity from orientation polarization, Deltaepsilon, of isoamylbromide is about the same as in the pure state, but that of 1-propanol decreases by a factor of approximately 3 at 120 K. As the temperature is decreased, this difference remains constant. The relaxation time, tau, of isoamylbromide and its distribution parameter remains the same as for the pure liquid, but that of 1-propanol is longer and increases with decrease in T, becoming approximately 130 times the pure liquid's value at 119 K. This is in contrast to the finding for an isomeric heptanol, whose tau had decreased. Extrapolation suggests that at T>151 K, tau of 1-propanol in the mixture may become less than that in the pure liquid (the isoamylbromide component crystallizes before this temperature could be reached). This indicates that Tg corresponding to tau of 10(3) s for 1-propanol in the mixture would be higher than in the pure liquid. Crystallization of the two components in the mixture occurs at different rates and 1-propanol remains partially uncrystallized while isoamylbromide completely crystallizes. tau of any remaining liquid isoamylbromide does not change in the presence of crystallized states while tau of residual liquid 1-propanol in the mixture is reduced. The mixture phase separates in submicron or nanosize aggregates of the alcohol in isoamylbromide, without affecting the latter's relaxation kinetics, while its own epsilon(s) decreases and tau increases. Consequences of the finding for various relaxation mechanisms are briefly described.

Relaxations and nano-phase-separation in ultraviscous heptanol-alkyl halide mixture

To gain insight into the effects of liquid-liquid phase separation on molecular relaxation behavior we have studied an apparently homogeneous mixture of 5-methyl-2-hexanol and isoamylbromide by dielectric spectroscopy over a broad temperature range. It shows two relaxation regions, widely separated in frequency and temperature, with the low-frequency relaxation due to the alcohol and the high-frequency relaxation due to the halide. In the mixture, the equilibrium dielectric permittivity epsilon(s) of the alcohol is 41% of the pure state at 155.7 K and epsilon(s) of isoamylbromide is approximately 86% of the pure state at 128.7 K. The difference decreases for the alcohol component with decreasing temperature and increases for the isoamylbromide component. The relaxation time tau of 5-methyl-2-hexanol in the mixture at 155.7 K is over five orders of magnitude less than in the pure state, and this difference increases with decreasing temperature, but tau of isoamylbromide in the mixture is marginally higher than in the pure liquid. This shows that the mixture would have two T(g)'s corresponding to its tau of 10(3) s, with values of approximately 121 K for its 5-methyl-2-hexanol component and approximately 108 K for its isoamylbromide component. It is concluded that the mixture phase separates in submicron or nanometer-size aggregates of the alcohol in isoamylbromide, without affecting the latter's relaxation kinetics, while its own epsilon(s) and tau decrease markedly.

The Crystallization Behavior and Mechanical Properties of Poly(ethylene-ran-vinyl acetate) and Paraffin Wax Blend.

The effect of liquid-liquid phase-separation (LLPS) on the crystallization behavior and mechanical properties of poly(ethylene-ran-vinyl acetate) (EVA) and paraffin wax blend was investigated. The blend of EVA and the wax became homogeneous at temperatures greater than its upper critical solution temperature (UCST) (371K), and an LLPS was observed between UCST and the melting point of 361 K for EVA in the blend. The degree of crystallinity of EVA in the blend, judged from a melting endothermic peak in differential scanning calorimeter (DSC) thermograms obtained during heating runs, decreased with increasing duration time in the LLPS region.The flexural modulus of the blend became maximum at the 35wt% EVA. This behavior can be explained by the fact that this blend composition has the largest relative degree of crystallinity of EVA measured by DSC. We found that the flexural modulus of the binder itself is directly related to that of a feedstock consisting of larger amounts of metal powder and the binder, which can help someone to develop a suitable binder system for a powder injection molding process.

Phase behaviour of poly(ϵ-caprolactone)/(polystyrene- ran-acrylonitrile) blends exhibiting both liquid-liquid unmixing and crystallization

Miscibility and morphology in blends of poly(ϵ-caprolactone) (PCL) and poly(styrene- ran-acrylonitrile) (SAN) were examined by light scattering and optical microscopy. The blends display miscibility in the amorphous state in a limited range of copolymer composition. Morphologies, formed as a combined effect of liquid-liquid phase separation and crystallization of PCL, are discussed. Crystallization leads to spherulite structure in the blends. Upon crystallization, PCL segregates from the amorphous state resulting in the observation of ring patterns in miscible blends. Its periodicity decreases with increasing fraction of SAN in the blend. Ring-banded spherulites have not been observed in neat PCL or in immiscible blends. These effects are discussed in terms of twisted crystallization.

Gels of poly(vinyl alcohol) from dimethyl sulphoxide/water solutions

Sol-gel diagrams of atactic poly(vinyl alcohol) (at-PVA) in a mixture of dimethyl sulphoxide and water (60/40 vol/vol) have been investigated. The crosslinking points of the gels are crystallites of PVA. Gelation of seven samples of at-PVA having degrees of polymerization ( P n) ranging from 186 to 20 200 was observed at quenching temperatures ( T Q s ) between −60°C and 80°C. Gelation time, which is defined as time required for gelation, decreases with decreasing T Q. In a temperature range below ∼ −20°C, the gels are transparent and the critical gelation concentrations ( C∗ s ) are independent of T Q for all the samples except the lowest P n. Further, C∗ s in this temperature range are proportional to P −0.5 n, suggesting that C∗ is determined by the polymer chain overlap concentration. On the other hand, gels formed above −20°C are not transparent, and the degree of transparency decreases with increasing T Q. The lowest C∗ which is lower than the overlap concentration for polymer chains with radius of gryration R g is observed in the T Q range of −20 to 40°C. This may be explained by the effect of liquid-liquid phase separation or spinodal decomposition, which is supported by T Q and PVA concentration dependences of u.v. transmission.