Ice Nucleation Research Articles

Biophysical evidence that frostbite is triggered on nanocrystals of biogenic magnetite in garlic cloves (Allium sativum)

Trace levels of biologically precipitated magnetite (Fe3O4) nanocrystals are present in the tissues of many living organisms, including those of plants. Recent work has also shown that magnetite nanoparticles are powerful ice nucleation particles (INPs) that can initiate heterogeneous freezing in supercooled water just below the normal melting temperature. Hence there is a strong possibility that magnetite in plant tissues might be an agent responsible for triggering frost damage, even though the biological role of magnetite in plants is not understood. To test this hypothesis, we investigated supercooling and freezing mortality in cloves of garlic (Allium sativum), a species which is known to have moderate frost resistance. Using superconducting magnetometry, we detected large numbers of magnetite INPs within individual cloves. Oscillating magnetic fields designed to torque magnetite crystals in situ and disturb the ice nucleating process produced significant effects on the temperature distribution of supercooling, thereby confirming magnetite’s role as an INP in vivo. However, weak oscillating fields increased the probability of freezing, whereas stronger fields decreased it, a result that predicts the presence of magnetite binding agents that are loosely attached to the ice nucleating sites on the magnetite crystals.

From foes to friends; bacterial proteins for optimal wart cryotherapy

Warts are a common skin complaint caused by viral infections. Cryotherapy is a well-established approach for wart removal, which apply low temperatures to freeze and destroy infectious cells. However, this process may face-up several unwanted side effect on healthy nearby tissues. Besides, proper penetration of such temperature is also required to cause irreversible damages in deep-root infectious layers. In this respect, ice-producing proteins that are naturally found in some bacterial species, knowing as ice nucleation active (INA) bacteria, may be practical. In the current work, we hypothesized purification and encapsulation of these proteins into vesicular lipid particles (such as liposomes, niosomes, and exosomes) for an intralesional injection. Thereby, effective wart removal would be achieved even at low temperatures without undesirable effects.

Inverse Relationship Between Ice Nucleation and Ice Growth Rates in Frozen Foods

Inverse Relationship Between Ice Nucleation and Ice Growth Rates in Frozen Foods

The role of the water contact layer on hydration and transport at solid/liquid interfaces

Understanding the structure in the nanoscopic region of water that is in direct contact with solid surfaces, so-called contact layer, is key to quantifying macroscopic properties that are of interest to e.g. catalysis, ice nucleation, nanofluidics, gas adsorption, and sensing. We explore the structure of the water contact layer on various technologically relevant solid surfaces, namely graphene, MoS[Formula: see text], Au(111), Au(100), Pt(111), and Pt(100), which have been previously hampered by time and length scale limitations of ab initio approaches or force field inaccuracies, by means of molecular dynamics simulations based on ab initio machine learning potentials built using an active learning scheme. Our results reveal that the in-plane intermolecular correlations of the water contact layer vary greatly among different systems: Whereas the contact layer on graphene and on Au(111) is predominantly homogeneous and isotropic, it is inhomogeneous and anisotropic on MoS[Formula: see text], on Au(100), and on the Pt surfaces, where it additionally forms two distinct sublayers. We apply hydrodynamics and the theory of the hydrophobic effect, to relate the energy corrugation and the characteristic length-scales of the contact layer with wetting, slippage, the hydration of small hydrophobic solutes and diffusio-osmotic transport. Thus, this work provides a microscopic picture of the water contact layer and links it to macroscopic properties of liquid/solid interfaces that are measured experimentally and that are relevant to wetting, hydrophobic solvation, nanofluidics, and osmotic transport.

Impact of controlled ice nucleation on intracellular dehydration, ice formation and their implications on T cell freeze–thaw viability

Cryopreservation is important in manufacturing of cell therapy products, influencing their safety and effectiveness. During freezing and thawing, intracellular events such as dehydration and ice formation can impact cell viability. In this study, the impact of controlling the ice nucleation temperature on intracellular events and viability were investigated. A model T cell line, Jurkat cells, were evaluated in commercially relevant cryoformulations (2.5 and 5 % v/v DMSO in Plasma-Lyte A) using a cryomicroscopic setup to monitor the dynamic changes cells go through during freeze–thaw as well as a controlled rate freezer to study bulk freeze–thaw. The equilibrium freezing temperatures of the studied formulations and a DMSO/Plasma-Lyte A liquidus curve were determined using DSC. The cryomicroscopic studies revealed that an ice nucleation temperature of −6°C, close to the equilibrium freezing temperatures of cryoformulations, led to more intracellular dehydration and less intracellular ice formation during freezing compared to either a lower ice nucleation temperature (−10 °C) or uncontrolled ice nucleation. The cell membrane integrity and post thaw viability in bulk cryopreservation consistently demonstrated the advantage of the higher ice nucleation temperature, and the correlation between the cellular events and cell viability.

Effect of salt-induced supercooling stability on the quality of chicken breast during supercooled storage

The effect of salt on the stability and quality of fresh chicken breast meat during supercooling preservation was investigated. The results showed that 0.75% salt kept the non-frozen state of chicken breast at −3 °C for 12 days. Compared to storage at −1 °C, storing at −3 °C could further extend the shelf life of chicken, but there was a risk of freezing the chicken meat (31% frozen after 4 d and all after 8 d). Microstructure images revealed that muscle fibers in the −3 °C samples were destroyed by crystallization, whereas ice nucleation was not observed in the −3 °C + salt treatment. The drip loss and centrifuging loss of the −3 °C + salt group after 12 days of storage (1.48%, 20.10%) were significantly lower than those of the −1 °C group (4.14%, 24.49%), the −3 °C group (8.37%, 27.47%), and the −1 °C + salt group (1.99%, 22.60%). Low-field nuclear magnetic resonance results showed that water migration in chicken breast was inhibited with −3 °C + salt treatment. Overall, the good quality of chicken breast was maintained by −3 °C + salt. The results provide a theoretical basis for industrial application of supercooling preservation of chicken.

Anti-corrosion and anti-icing properties of superhydrophobic laser-textured aluminum surfaces

Superhydrophobic surfaces have favorable properties in simultaneously reducing the negative effects of corrosion and ice accumulation. In this study, laser-texturing was employed as a facile and environmentally friendly surface method to prepare a surface with hierarchical roughness for subsequent grafting by immersion in an ethanol solution containing 1H,1H,2H,2H-perfluorodecyltriethoxysilane (FAS-10). Various analytical techniques were utilized to assess the characteristics of the laser-textured aluminum surfaces before and after grafting, such as a contact profilometer, optical tensiometer, scanning electron microscope with energy-dispersive spectroscope, and X-ray photoelectron spectroscope. These methods were used to evaluate surface roughness, wettability, morphology, and composition. The corrosion properties were evaluated through potentiodynamic and impedance measurements in a dilute Harrison's solution (DHS) composed of 0.35 wt% (NH4)2SO4 + 0.05 wt% NaCl. Additionally, freezing delay tests at various surface temperatures were performed to assess the surface's ability to prevent the freezing of water droplets on the treated surface. The laser-textured aluminum surface, featuring micro/nanostructures and a grafted nanoscopic perfluoroalkyl silane film, exhibited outstanding superhydrophobicity and enhanced corrosion protection. The developed surface has been shown to significantly delay the onset of ice nucleation and extend the freezing delay.

Rapid communication: A Simple, Non-Invasive, Cost-Effective Technique to Monitor Ice Nucleation in GMP Freeze-Drying

Rapid communication: A Simple, Non-Invasive, Cost-Effective Technique to Monitor Ice Nucleation in GMP Freeze-Drying

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.

Alternate Chemical Compounds as a Condensation Nucleus in Cloud Seeding

Cloud seeding involves boosting precipitation by releasing substances into the air that act as cloud condensation or ice nuclei. These substances encourage the development of clouds and precipitation. It’s like giving Mother Nature a gentle push to assist with rainfall in specific areas. The current work aimed to suggest Al2O3 as an alternate compound in cloud seeding rather than silver iodide. In this research, a unique approach is used to identify condensation nuclei, which play a crucial role in cloud formation and droplet growth. Various samples and four sources were included in the current study; refrigerated helfa powder, Himalayan salt, generator powder, and pollen, were analyzed using different physicochemical instruments. The proportions of chemical compounds in the samples show that there is 1.392% of Al2O3 in Refrigerated helfa which is the highest than in the other 3 sources, while the proportions of elements in the samples indicate that refrigerated helfa contains the lowest toxic compound, and although Al2O3 is insoluble in water, it is hygroscopic and can absorb 6.4% of humidity within 24 hours. As for the surface tension, refrigerated helfa shows lower density and surface tension than the other three sources with values of 0.9480 and 47.89 respectively. Al2O3 shows high humid absorptivity and refrigerated helfa can be used as a main source for Al2O3 which has a low effect on biota and is recommended for use in cloud seeding. However further work is recommended to be carried out in using Al2O3 as an alternative compound to silver iodide in cloud seeding.

Effects of Turbulence on Upper-Tropospheric Ice Supersaturation

Abstract Effects of turbulence on ice supersaturation at cirrus heights (>8 km) remain unexplored. Small-scale mixing processes become important for high Reynolds number flows, which may develop below the buoyancy length scale (10–100 m). The current study couples a stochastic turbulent mixing model with reduced dimensionality to an entraining parcel model to investigate, in large-ensemble simulations, how supersaturation evolves due to homogeneous turbulence in the stably stratified, cloud-free upper troposphere. The rising parcel is forced by a mesoscale updraft. The perturbation of an initially homogeneous vertical distribution of supersaturation is studied after a 36-m ascent in a baseline case and several sensitivity scenarios. Turbulent mixing and associated temperature fluctuations alone lead to changes in ensemble-mean distributions with standard deviations in the range 0.001–0.006, while mean values are hardly affected. Large case-to-case variability in the supersaturation field is predicted with fluctuation amplitudes of up to 0.03, although such large values are rare. A vertical gradient of supersaturation (≈10−3 m−1) is generated for high turbulence intensities due to the development of a dry-adiabatic lapse rate. Entrainment of slightly warmer (less than 0.1 K) environmental air into the parcel decreases the mean supersaturation by less than 0.01. Supersaturation fluctuations are substantially larger after entrainment events with an additional small offset in absolute humidity (by ±3.5%) between the parcel and environmental air. The predicted perturbations of ice supersaturation are significant enough to motivate studies of turbulence–ice nucleation interactions during cirrus formation that abandon the assumption of instantaneous mixing inherent to traditional parcel models. Significance Statement The purpose of our study is to investigate the effects of microscale turbulence on ice supersaturation in the upper troposphere. The associated variability in temperature and moisture fields is not resolved in cloud models and cannot easily be represented in terms of large-scale flow variables. We specify the conditions in which turbulent mixing and entrainment cause substantial variations in distributions of supersaturation. These include high turbulence intensity, strong atmospheric stability, and large moisture gradients. Our results suggest that turbulence may affect the strongly supersaturation-dependent ice formation processes in high-altitude clouds, pointing to the need to investigate cirrus formation in the presence of turbulence.

Ice nucleation in supercooled water under shear

Classical nucleation theory (CNT) provides a good framework for understanding the nucleation phenomenon in metastable conditions. However, there is still no satisfactory understanding of the shear effect. Here, we propose an ice nucleation model considering the rebounded deformation energy to analyze the ice nucleation rate. It is concluded that: (1) The effect is more pronounced by changing the contact angle to regulate the free energy when both the supercooled degree and the shear rate are small. (2) The variation trend of the nucleation ratio Jhet,sh/Jhet,no increases at first but it may decrease when the contact angle is small, or keeps increasing when the contact angle is large. (3) The probability of the shear nucleation is about several ten orders of magnitude to that of static nucleation. (4) The key factor in the acceleration of the supercooled liquid nucleation is the released deformation energy after encountering the obstruction.

Nucleation mechanism in the absence and presence of an electric field

The objective of this article is to investigate the mechanism of the ice nucleation process theoretically by assaying the thermodynamics of the ice nucleus as a small system, and experimentally verified by studying the effect of the static electric field on the ice nucleation process. Using the thermodynamic of small system related to Hill, we predicted that the equilibrium temperature of water and ice nucleus decreases with the reduction of nucleus size. By applying an electrostatic field, the Gibbs free energy of nucleus formation decreases due to this fact that the electrostatic work on the ice nucleus is lower than that for bulk water which it causes a decrease in the critical size of the ice nucleus. The experimental results showed that the nucleation temperature of water increases by applying the electric field with the voltage of 4.5 kV (internal strength of 2.2 × 104 V/m) while the size of ice crystals decreases in the frozen agar gel at the presence of electric field.

Role of the structural order of the hydration layer in regulating the heterogeneous ice nucleation efficiency

Understanding the complex microscopic mechanisms of heterogeneous ice nucleation is crucial across diverse fields from cloud science to microbiology. In this work, we examined the ability of solid surfaces to promote ice nucleation as a function of the structural order of interfacial water, including the first and second hydration layers. Comprehensive all-atom molecular dynamics simulations using the TIP4P/ice water model were conducted on a water film atop a modeled surface inspired by the (0001) surface of AgI crystal, with varying surface charges. These simulations revealed non-monotonic nucleation rates. The first hydration layer formed a hexagonal hydrogen-bond network resembling ice structures, exhibiting low mobility essential for inducing ice nucleation. Notably, the second hydration layer, acting as a key-point bridge, displayed varying degrees of orientational preference facilitated by inter-layer hydrogen bonds. The calculation of fluctuation parameters indicated that the greater structural order of the second layer significantly enhances the surface’s ice-forming capabilities. Unlike previous studies that focused primarily on the role of the first layer of interfacial water, our findings demonstrate that the second layer provides a more accurate indicator of ice nucleation capabilities.

Bridging classical nucleation theory and molecular dynamics simulation for homogeneous ice nucleation.

Water freezing, initiated by ice nucleation, occurs widely in nature, ranging from cellular to global phenomena. Ice nucleation has been experimentally proven to require the formation of a critical ice nucleus, consistent with classical nucleation theory (CNT). However, the accuracy of CNT quantitative predictions of critical cluster sizes and nucleation rates has never been verified experimentally. In this study, we circumvent this difficulty by using molecular dynamics (MD) simulation. The physical properties of water/ice for CNT predictions, including density, chemical potential difference, and diffusion coefficient, are independently obtained using MD simulation, whereas the calculation of interfacial free energy is based on thermodynamic assumptions of CNT, including capillarity approximation among others. The CNT predictions are compared to the MD evaluations of brute-force simulations and forward flux sampling methods. We find that the CNT and MD predicted critical cluster sizes are consistent, and the CNT predicted nucleation rates are higher than the MD predicted values within three orders of magnitude. We also find that the ice crystallized from supercooled water is stacking-disordered ice with a stacking of cubic and hexagonal ices in four representative types of stacking. The prediction discrepancies in nucleation rate mainly arise from the stacking-disordered ice structure, the asphericity of ice cluster, the uncertainty of ice-water interfacial free energy, and the kinetic attachment rate. Our study establishes a relation between CNT and MD to predict homogeneous ice nucleation.

Sub-zero non-freezing of vascularized composite allografts in a rodent partial hindlimb model

Ischemia is a major limiting factor in Vascularized Composite Allotransplantation (VCA) as irreversible muscular injury can occur after as early as 4–6 h of static cold storage (SCS). Organ preservation technologies have led to the development of storage protocols extending rat liver ex vivo preservation up to 4 days. Development of such a protocol for VCAs has the added challenge of inherent ice nucleating factors of the graft, therefore, this study focused on developing a robust protocol for VCA supercooling. Rodent partial hindlimbs underwent subnormothermic machine perfusion (SNMP) with several loading solutions, followed by a storage solution with cryoprotective agents (CPA) developed for VCAs. Storage occurred in suspended animation for 24h and VCAs were recovered using SNMP with modified Steen. This study shows a robust VCA supercooling preservation protocol in a rodent model. Further optimization is expected to allow for its application in a transplantation model, which would be a breakthrough in the field of VCA preservation.

Cryo-Nanocatalyst Enhances Therapeutic Efficacy of Cryo-Immunotherapy through Necroptosis and Local Delivery of Programmed Death-Ligand 1 Inhibitors.

Combining cryoablation and immunotherapy presents a promising approach to revert immunosuppressive responses to solid tumors. However, challenges such as postablated residual tumors and insufficient immune activity contribute to recurrence after cryo-immunotherapy. Herein, we investigated metallic supra-structured cryo-nanocatalyst (MSCN), which features numerous ice nucleation sites and interspace loading of therapeutic agents. MSCN elevates the freezing point and enhances ice nucleation, facilitating effective ice formation during cryotreatment. MSCN-loaded tumor cells showed a 2-fold increase in cryo-cytotoxicity and undergo osmotic-related cell damage, primarily necroptosis rather than other regulated cell death mechanisms. In prostate cancer models, RNA sequencing reveals that MSCN-cryoablation promoted antitumor inflammatory pathways, including necroptosis, compared to cryoablation alone. Additionally, following programmed death-ligand 1 (PD-L1) upregulation postcryoablation, synergistic effects with PD-L1 blockade were confirmed. Given the interspace of MSCN for aPD-L1 loading, we compared the intratumoral delivery of PD-L1 blockade against systemic injection. Enhanced necrosis and necroptosis from MSCN-cryoablation and PD-L1 blockade effectively eradicated tumors and triggered antitumor and memory immune responses locally and systemically. Lastly, a spatial landscape of tumor-infiltrating immune cells was analyzed to gain insight into heterogeneous tumor responses, leading to the limitations of conventional focal ablation techniques. Our findings highlight the potential of advanced cryo-immunotherapy using cryo-nanocatalysis to promote ice formation and necroptosis, stimulating antitumor immunogenic responses.

Fungal aerosols over the Northwest Pacific to Arctic Ocean: The influence of air mass and environmental factors

Bioaerosols, prevalent in the atmosphere, serve as carriers for microbe transmission and can also act as Cloud Condensation Nuclei (CCN) and Ice Nuclei (IN), exerting significant influence on cloud microphysics, radiative forcing, precipitation processes, and global climate dynamics. In this study, we conducted a comprehensive study of airborne fungi from the Northwest Pacific to the Arctic Ocean. During the 9th Chinese National Arctic Research Expedition (Chinare, 2018) cruise, total suspended particulate matter (TSP) samples were collected, followed by high-throughput sequencing to determine the microbial community's structure. Ascomycota and Basidiomycota are widespread in marine fungal aerosols along this route, with a notably higher abundance of Epicoccum and Cladosporium in fungal aerosols from the low-latitude Northwest Pacific compared to the high-latitude Arctic Ocean region. The composition of fungal communities depended on air mass transport processes and environmental factors. Long-range transport of terrestrial air masses not only enriches the abundance of fungal aerosols but also increases the relative abundance of dominant genera such as Basidiomycota, Epicoccum and Cladosporium. The knowledge gap about fungal aerosols over the Arctic route from the Northwest Pacific to the Arctic Ocean was filled in part by this work. This study deepens our understanding of how environmental factors and terrestrial air masses influence the composition and distribution of fungal community, offering valuable insights into the interconnections between marine and terrestrial ecosystems and their potential impacts on climate and atmospheric processes.

Microplastic Particles Contain Ice Nucleation Sites That Can Be Inhibited by Atmospheric Aging.

Recent research has shown that microplastics are widespread in the atmosphere. However, we know little about their ability to nucleate ice and their impact on ice formation in clouds. Ice nucleation by microplastics could also limit their long-range transport and global distribution. The present study explores the heterogeneous ice-nucleating ability of seven microplastic samples in immersion freezing mode. Two polypropylene samples and one polyethylene terephthalate sample froze heterogeneously with median freezing temperatures of -20.9, -23.2, and -21.9 °C, respectively. The number of ice nucleation sites per surface area, ns(T), ranged from 10-1 to 104 cm-2 in a temperature interval of -15 to -25 °C, which is comparable to that of volcanic ash and fungal spores. After exposure to ozone or a combination of UV light and ozone, simulating atmospheric aging, the ice nucleation activity decreased in some cases and remained unchanged in others. Our freezing data suggest that microplastics may promote ice formation in cloud droplets. In addition, based on a comparison of our freezing results and previous simulations using a global transport model, ice nucleation by microplastics will impact their long-range transport to faraway locations and global distribution.

Polyurethane Nanocomposite Coatings Coupled with Titanium-Based Conversion Layers for Enhanced Anticorrosion, Icephobic Properties, and Surface Protection.

This study examines the efficacy of icephobic polyurethane nanocomposite coatings in mitigating corrosion on an aluminum substrate. A titanium-based conversion coating is applied to modify the substrate, and the research focuses on optimizing the dual functionalities of icephobicity and anticorrosion within the polyurethane coatings while ensuring strong substrate adhesion. The coatings are formulated using fluoropolyol, isocyanate, and silica nanoparticles treated with polydimethylsiloxane. Surface properties are analyzed using contact angles, contact angle hysteresis measurements, and atomic force microscopy, and the coatings' icephobicity is evaluated through differential scanning calorimetry, freezing time delay, ice adhesion under impact and non-impact conditions, and ice accretion tests. The corrosion resistance and adhesive strength of the coatings are assessed using electrochemical impedance spectroscopy and cross-cut tests, respectively. Increasing the concentration of silica nanoparticles to 10 wt.% increases contact angles to 167°, although the 4 wt.% coating produces the lowest contact angle hysteresis (3° ± 0.5°) and ice nucleation temperature (-23 °C). The latter coating is then applied to a substrate pretreated with a titanium/cerium-based conversion coating. This prepared surface maintains an ice adhesion of about 15 kPa after 15 icing/de-icing cycles and provides approximately 90 days of surface protection (|Z|lf = 1.6 × 109 Ω·cm2). Notably, the impedance value exceeds that of untreated substrates, underscoring the effectiveness of the titanium/cerium-based conversion coating in enhancing both corrosion resistance and coating adhesion to the substrate.