Ice Crystal Surface Research Articles

Ice/salt assisted synthesis of ultrathin two-dimensional nitrogen doped carbon nanosheets for fast lithium storage

Two-dimensional carbon nanosheets have garnered considerable interest as promising anode materials in energy storage devices due to their large specific surface area and excellent conductivity. The efficient synthesis of ultrathin nitrogen-doped carbon nanosheets is achieved with the aid of cysteine, employing a green ice/salt-assisted strategy. Ice influences the growth direction of salt crystals and determines the distribution of sodium lignosulfonate on the surfaces of both ice and salt crystals. The introduction of methionine incorporates heteroatoms, altering the electronic structure and chemical activity of the material, thereby effectively enhancing its lithium storage performance. The initial charge capacity of the optimized sample reached 766.4 mA h g−1 at a current density of 0.2 A g−1, maintaining a capacity of 571.1 mA h g−1 after 100 cycles. This study presents an innovative method for fabricating ultrathin carbon nanosheets using eco-friendly, cost-effective sodium lignosulfonate, showcasing a promising candidate for lithium-ion batteries and future energy storage technologies.

Composition-antifreeze property relationships of gelatin and the corresponding mechanisms

The inherent functional fractions (gelation and ice-affinitive fractions) of gelatin enable it as a promising cryoprotectant alternative. However, the composition-antifreeze property relationships of gelatin remain to be investigated. In this study, the HW-PSG and LW-PSG fractions of gelatin from fish scales were obtained, according to the critical gelation conditions and ice-binding measurements, respectively. Thermal hysteresis (THA) value, associated with ice nucleation, of LW-PSG was higher than that of HW-PSG. Besides, the relatively low-sized ice crystals (210–550 μm2) indicated that HW-PSG showed strong ice recrystallization inhibition (IRI) ability, compared to other groups. These results suggested that LW-PSG inhibited ice nucleation, while HW-PSG displayed the strong IRI ability. Furthermore, the antifreeze mechanisms were clarified through IRI measurements and molecular dynamics simulation. The minimum size of ice crystals was found for HW-PSG gels with dense microstructure, suggesting the HW-PSG retarded the growth of ice crystals by restricting the migration and phase transformation of water molecules. The hydrogen bond interactions between the ice crystal surface and ASN1294 and PRO1433 residues of LW-PSG, and hydrophobic interactions contributed to inhibiting the nucleation of ice crystals. This study provided some references to further enhance antifreeze performance of gelatin by modulating fragment composition.

Prediction of heat and mass transfer inside 3D frost microstructure

In the present study, a CFD model based on 3D frost microstructure is developed. The desublimation phase change coupled with mass transfer of water vapor to ice surface and heat conduction through the tortuous ice crystals are calculated to predict accurate heat transfer process inside the frost microstructure. Air velocity and temperature inside the frost structure after 30 min operation are significantly lower than those inside 10 min operation, which are influenced by the structural change of the ice crystals. Vertical variations in ice crystal surface temperature and ice surface mass flux are also calculated, which varied between the frost structures operated for different periods. At 70 μm height of the frost, the mass flux at the ice crystal surface shows significant decrease from 7.48 × 10−5 kg/m2s for the frost structure at 10 min to 1.82 × 10−5 kg/m2s for that at 30 min. This study provides new and fundamental information to understand the mechanism of frost growth which is very important in refrigeration, cryogenics, and aerospace industry, etc.

Pore-scale physics of ice melting within unconsolidated porous media revealed by non-destructive magnetic resonance characterization

Melting of ice in porous media widely exists in energy and environment applications as well as extraterrestrial water resource utilization. In order to characterize the ice-water phase transition within complicated opaque porous media, we employ the nuclear magnetic resonance (NMR) and imaging (MRI) approaches. Transient distributions of transverse relaxation time T2 from NMR enable us to reveal the substantial role of inherent throat and pore confinements in ice melting among porous media. More importantly, the increase in minimum T2 provides new findings on how the confinement between ice crystal and particle surface evolves inside the pore. For porous media with negligible gravity effect, both the changes in NMR-determined melting rate and our theoretical analysis of melting front confirm that conduction is the dominant heat transfer mode. The evolution of mushy melting front and 3D spatial distribution of water content are directly visualized by a stack of temporal cross-section images from MRI, in consistency with the corresponding NMR results. For heterogeneous porous media like lunar regolith simulant, the T2 distribution shows two distinct pore size distributions with different pore-scale melting dynamics, and its maximum T2 keeps increasing till the end of melting process instead of reaching steady in homogeneous porous media.

Ice Recrystallization Unveils the Binding Mechanism Operating at a Diffused Interface.

Recrystallization of ice is a natural phenomenon that causes adverse effects in cryopreservation, agriculture, and in frozen food industry. It has long been recognized that ice recrystallization occurs through the Ostwald ripening and accretion processes. However, neither of these processes has been explored in microscopic detail by state-of-the-art experimental techniques. We carried out atomistic molecular dynamics (MD) simulations to explore ice recrystallization through the accretion process. Attempts have been made to elucidate the binding mechanism that is operating at the diffused ice-water interface. It is demonstrated that two ice crystals spontaneously recognize each other and bind together to form a large crystal in liquid water, resulting in ice recrystallization by accretion. Interestingly, the study reveals that the binding occurs due to the freezing of the interfacial water layer present between the two ice planes, even at a temperature above the melting point of the ice crystal. The synergistically enhanced ordering effect of two ice surfaces on the interfacial water leads to such freezing occurring during the binding process. However, proper crystallographic alignment is not necessarily required for the binding of the two crystals. Simulations have also been carried out to study the binding between an ice crystal and the model ice-binding surface (IBS) of an antifreeze protein above the melting point of the ice crystal. It is found that such binding at the IBS is accompanied by freezing of the interfacial water. This establishes that the synergetic ordering-driven freezing of interfacial water is a common binding mechanism at the diffused surfaces of ice crystals. We believe that this mechanism will provide a microscopic understanding of the process of recrystallization inhibition and thus help in designing suitable materials for potent applications in recrystallization inhibition.

In Situ Transmission Electron Microscopy Study of Bubble Behavior Near the Surface of Ice Crystals by Using a Liquid Cell With a Peltier Cooling Holder.

Liquid cell transmission electron microscopy (LC-TEM) is a unique technique that permits in situ observations of various phenomena in liquids with high spatial and temporal resolutions. One difficulty with this technique is the control of the environmental conditions in the observation area. Control of the temperature ranging from room temperature to minus several tens of degrees Celsius, is desirable for controlling the supersaturation in various materials and for observing crystallization more easily. We have developed a cooling transmission electron microscopy specimen holder that uses Peltier devices, and we have combined it with a liquid cell to realize accurate temperature control in LC-TEM. We evaluated this system by using water as a specimen. Motionless bubbles, shown to be voids containing pressurized gas, formed in the specimen sometime after the temperature had reached -12°C. An electron diffraction pattern showed that the specimen turned into ice Ih after the formation of these bubbles, confirming that our system works properly and can induce crystallization. In addition, we analyzed the behavior of bubbles formed in the ice Ih, and we discussed the formation of these bubbles and their internal pressure.

Characterization and the mechanism underlying the cryoprotective activity of a peptide from large yellow croaker (Pseudosciaena crocea)

Ice crystal-induced protein denaturation is the main cause of the deterioration of fish during frozen storage and transportation. In this study, the ultra-performance liquid chromatography − quadrupole − time of flight (UPLC-Q-TOF) technique was used to identify and screen tryptic peptides Ile-Glu-Glu-Leu-Glu-Glu-Leu-Glu-Ala-Glu-Arg (IEELEEELEAER) from large yellow croaker (Pseudosciaena crocea). The results were used study their cryoprotective effects on turbot fish meat during freeze–thaw cycles at different concentrations, and to investigate their anti-freezing mechanism. The results showed that the I-2.0 group effectively inhibiting the degeneration and structure changes of myofibrillar proteins after three freeze–thaw cycles, and the Ca2+-ATPase activity (1.65 μmolPi/mg/h), increased by 55.86% compared with that of the control group. Additionally, peptide IEELEEELEAER could provide antifreeze protection by binding to the surface of ice crystals and inhibiting their transformation. This peptide acts as a natural cryoprotectant and might be used for the cryogenic storage and transportation of fish products.

Adsorption Kinetics of Type-III Antifreeze Protein on Ice Crystal Surfaces.

The spatio-temporal distribution of type-III antifreeze protein (AFP-III) molecules labeled with fluorescent isocyanate (FITC) was visualized at the interfaces between ice and solutions with an FITC-labeled AFP-III (F-AFP-III) concentration of 20-800 μg/mL by fluorescence microscopy. The number density of F-AFP-III on the surface of ice microcrystals was calculated from the calibrated fluorescence intensity. The adsorption of F-AFP-III molecules on the ice crystal surfaces proceeded at a finite rate and then reached the saturation level. The time course of the number density of adsorbed F-AFP-III molecules could be well represented by Langmuir's model. The characteristic adsorption time of F-AFP-III, the adsorption coefficient k1 = (0.5 ± 0.05) × 10-4 (μg/mL)-1 s-1, and the desorption coefficient k2 = 0.005 ± 0.002 s-1 were determined using the Langmuir's model and obtained experimental data. We found that the adsorption of F-AFP-III could have different kinetics depending on the solution conditions and the type of fluorescence molecules conjugated with AFP-III.

Oligonucleotides as Inhibitors of Ice Recrystallization.

Oligonucleotides of adenine (A20), guanine (G20), cytosine (C20), thymine (T20), cytosine-guanine ((CG)20), and adenine-thymine ((AT)20) were investigated as model compounds for ice recrystallization inhibition (IRI). Dehydroxy uracil (dU20), U20, and T20 were also compared to investigate the effect of minute changes in the hydrophobicity of the oligonucleotides on the IRI activity. Among the oligonucleotides considered in this study, T20 exhibited the best performance for IRI. In addition, the degree of polymerization of oligothymines varied over 5, 10, 20, 30, 50, and 100, and T20 was found to be the most effective for IRI. The IRI mechanism was investigated by comparing U20 and T20, which exhibited the lowest and highest IRI activity, respectively, among the oligonucleotides for their dynamic ice-shaping, thermal hysteresis, and ice nucleation inhibition. Little or no dynamic ice-shaping activity and small thermal hysteresis were observed for both nucleotides. All of the findings suggest that not the ice-polymer adhesion but the hydrophobic interactions of T20 in the interface layer might interfere with the water deposition on the ice crystal surfaces and contribute to the IRI activity of the T20 oligonucleotide.

Depletion interaction may reduce ice recrystallization inhibition activity of cellulose nanocrystals (CNCs) at high concentrations

In ice cream, food hydrocolloids are commonly used as stabilizers for controlling ice recrystallization. Recent studies suggest that some stabilizers might bind to ice crystal surfaces and inhibit recrystallization. However, this ice-binding mechanism is challenged by the fact that stabilizers at high concentrations sometimes have reduced activity or even accelerate ice recrystallization. In this work, we determined the ice recrystallization inhibition (IRI) activity of cellulose nanocrystals (CNCs) at different concentrations in the 3.0% sucrose solution by the splat assay and in the 25.0% and 40.0% sucrose solutions by the sandwich assay. Reduced activity at high CNCs concentrations was observed in the sandwich assay. It was not linked to the phase separation or liquid crystal phase formation of CNCs but was caused by the accretion of nearby ice crystals and the forming of large ice crystals with irregular morphology and high aspect ratios. A new explanation based on the depletion-interaction-induced accretion of ice crystals was proposed. Our findings might resolve the contradiction between the ice-binding mechanism and the stabilizer concentration effect and add more support to the interfacial mechanism of ice recrystallization inhibition. They also pointed out the importance of selecting suitable stabilizer concentrations for screening IRI active materials and manufacturing frozen desserts.

Understanding the History of Two Complex Ice Crystal Habits Deduced From a Holographic Imager

AbstractThe sizes and shapes of ice crystals influence the radiative properties of clouds, as well as precipitation initiation and aerosol scavenging. However, ice crystal growth mechanisms remain only partially characterized. We present the growth processes of two complex ice crystal habits observed in Arctic mixed‐phase clouds during the Ny‐Ålesund AeroSol Cloud ExperimeNT campaign. First, are capped‐columns with multiple columns growing out of the plates' corners that we define as columns on capped‐columns. These ice crystals originated from cycling through the columnar and plate temperature growth regimes, during their vertical transport by in‐cloud circulation. Second, is aged rime on the surface of ice crystals having grown into faceted columns or plates depending on the environmental conditions. Despite their complexity, the shapes of these ice crystals allow to infer their growth history and provide information about the in‐cloud conditions. Additionally, these ice crystals exhibit complex shapes and could enhance aggregation and secondary ice production.

Nanoscopy of single antifreeze proteins reveals that reversible ice binding is sufficient for ice recrystallization inhibition but not thermal hysteresis

Antifreeze proteins (AFPs) bind ice to reduce freezing temperatures and arrest ice crystal ripening, making AFPs essential for the survival of many organisms in ice-laden environments and attractive as biocompatible antifreezes in many applications. While their activity was identified over 50 years ago, the physical mechanisms through which they function are still debated because experimental insights at the molecular scale remain elusive. Here, we introduce subzero nanoscopy by the design and incorporation of a freezing stage on a commercial super-resolution setup to resolve the interfacial dynamics of single AFPs with ice crystal surfaces. Using this method, we demonstrate irreversible binding and immobilization (i.e., pinning) of individual proteins to the ice/water interface. Surprisingly, pinning is lost and adsorption becomes reversible when freezing point depression activity, but not ice recrystallization inhibition, is eliminated by a single mutation in the ice-binding site of the AFP. Our results provide direct experimental evidence for the adsorption-inhibition paradigm, pivotal to all theoretical descriptions of freezing point depression activity, but also reveal that reversible binding to ice must be accounted for in an all-inclusive model for AFP activity. These mechanistic insights into the relation between interfacial interactions and activity further our understanding and may serve as leading principles in the future design of highly potent, biocompatible antifreezes with tunable affinity.

Polysaccharide-Derived Ice Recrystallization Inhibitors with a Modular Design: The Case of Dextran-Based Graft Polymers.

Ice recrystallization inhibitors inspired from antifreeze proteins (AFPs) are receiving increasing interest for cryobiology and other extreme environment applications. Here, we present a modular strategy to develop polysaccharide-derived biomimetics, and detailed studies were performed in the case of dextran. Poly(vinyl alcohol) (PVA) which has been termed as one of the most potent biomimetics of AFPs was grafted onto dextran via thiol-ene click chemistry (Dex-g-PVA). This demonstrated that Dex-g-PVA is effective in IRI and its activity increases with the degree of polymerization (DP) (sizes of ice crystals were 18.846 ± 1.759 and 9.700 ± 1.920 μm with DPs of 30 and 80, respectively) and fraction of PVA. By means of the dynamic ice shaping (DIS) assay, Dex-g-PVA is found to engage on the ice crystal surfaces, thus the ice affinity accounts for their IRI activity. In addition, Dex- g-PVA displayed enhanced IRI activity compared to that of equivalent PVA alone. We speculate that the hydrophilic nature of dextran would derive PVA in a stretch conformation that favors ice binding. The modular design can not only offer polysaccharides IRI activity but also favor the ice-binding behavior of PVA.

Atomic force microscopy imaging of ice crystal surfaces formed in aqueous solutions containing ice-binding proteins

Ice-binding proteins (IBPs) allow organisms to survive below the freezing point by modulating ice crystal growth. These proteins act by binding to ice surfaces, thus inhibiting ice growth. Until now, high-resolution imaging of ice growing in the presence of IBPs has not been possible. We developed a unique in-situ technique that enables atomic force microscopy (AFM) imaging of ice formation and growth in the ice-IBP system. The new technique enables controlling the growth of ice crystals under a strong and focused thermal gradient. We present images of ice crystals with sub-ten nanometer resolution. Ice was grown in the presence of two different IBPs that exhibit specific and unique structures. This development opens the path for fine elucidation of the interaction of IBPs with growing ice surfaces as well as with other frozen systems at unprecedented high resolution. Furthermore, with the exception of crystals growing in thin films, this is the first demonstration for imaging a growing crystal immersed in its own melt with AFM.

Initial ice growth control mechanism for CMC-Na in model systems

In this study, CMC-Na solutions with different DS and concentrations were frozen at −20 °C and −40 °C. At the macroscopic level, the nucleation temperature and nucleation start time were analysed by freezing curves, and the morphology, size, and quantity of ice crystals after recrystallization were captured dynamically at the microlevel. The results show that CMC-Na can effectively accelerate the nucleation process, shorten the phase transition time, and evenly promote the formation of numerous distributed small ice crystals, and there is no significant difference in the effect of CMC-Na at different freezing temperatures. At the microlevel, a high concentration of CMC-Na can inhibit the recrystallization process more effectively, probably because it is readily attached to the surface of ice crystals, and the reduction in surface tension increases the free energy, which prevents the growth of ice crystals. When CMC-Na is added to the ice cream system, it can reduce the nucleation temperature, increase the degree of supercooling, and promote the internal generation of abundant ice crystals. At the same annealing rate, CMC-Na can effectively inhibit the occurrence of recrystallization, which is consistent with the conclusion of the solution system.

Adsorption of ice-binding proteins onto whole ice crystal surfaces does not necessarily confer a high thermal hysteresis activity

Many psychrophilic microorganisms synthesize ice-binding proteins (IBPs) to survive the cold. The functions of IBPs are evaluated by the effect of the proteins on the nonequilibrium water freezing-point depression, which is called “thermal hysteresis (TH)”, and the inhibitory effect of the proteins on the growth of larger ice crystals, which is called “ice recrystallization inhibition (IRI)”. To obtain mechanical insight into the two activities, we developed a modified method of ice affinity purification and extracted two new IBP isoforms from Psychromyces glacialis, an Arctic glacier fungus. One isoform was found to be an approximately 25 kDa protein (PsgIBP_S), while the other is a 28 kDa larger protein (PsgIBP_L) that forms an intermolecular dimer. Their TH activities were less than 1 °C at millimolar concentrations, implying that both isoforms are moderately active but not hyperactive IBP species. It further appeared that both isoforms exhibit high IRI activity even at submicromolar concentrations. Furthermore, the isoforms can bind to the whole surface of a hemispherical single ice crystal, although such ice-binding was generally observed for hyperactive IBP species. These results suggest that the binding ability of IBPs to whole ice crystal surfaces is deficient for hyperactivity but is crucial for significant IRI activity.

The mechanism of influence of polyvinyl alcohol (9 kDa) on the formation of ice crystals in aqueous solutions

Preventing crystallization of the liquid phase during freeze-thawing of cells is one of the main problems that need to be solved for the successful preservation of biomaterial at low temperatures. One highly effective recrystallization inhibitor is polyvinyl alcohol (PVA). However, the mechanisms of its cryoprotective effect have not been finally elucidated. In particular, it is not clear which structural features contribute to the realization of the antirecrystallization properties of PVA in the region of its cryoprotective concentrations. The influence of PVA on solvent crystallization and structural rearrangements of associations of PVA molecules in phosphate-buffered saline (PBS) was experimentally investigated. Solutions of PVA (molecular weight 9 kDa) in PBS were studied by cryomicroscopy and fluorescence spectroscopy methods. It was shown that molecular associations of PVA in PBS undergo a rearrangement of about 0.5−1 wt%, which is accompanied by a change in their size and hydrophilic-hydrophobic properties. PVA also changes the morphological structure of ice upon cooling and prevents crystallization upon heating. It is suggested that the mechanism of the antirecrystallization activity of PVA may be due to the formation of its complexes with the surface of ice crystals.

The emergence of drop-type and thin-layer-type quasi-liquid layers on ice crystal surfaces and their thermodynamic origin

Ice crystals play crucially important roles in wide variety of natural phenomena on the earth’s surface. However, in-situ optical observation of quasi-liquid layers (QLLs), which governs properties of ice crystal surfaces at temperature close to 0 °C, has been an experimental challenge. In this review, we show our recent studies on the in-situ observation of QLLs formed by the surface melting of ice crystals at temperatures below 0 °C. We first show our advanced optical microscopy that can directly visualize individual elementary steps (0.4 nm in thickness) on ice crystal surfaces. Then we next demonstrate the emergence of two kinds of QLLs with different morphologies (droplets and thin layers) on basal and prism faces of ice crystals. We also determined the surface tension-to-shear viscosity ratio (the so-called characteristic velocity) of both QLLs, indicating that the drop-type and thin-layer-type QLLs are 20- and 200-times less fluidic than bulk water, respectively. In addition, we measured the contact angles of drop-type QLLs on ice crystal surfaces and found that the formation of the drop-type and thin-layer-type QLLs can be explained by the wetting transition of QLLs on ice crystal surfaces. Furthermore, we determined the water vapor pressure–temperature ranges, in which both QLLs can present, and found that both QLLs are formed kinetically only in supersaturated or undersaturated water vapor. Our findings show a novel picture of surface melting that were significantly different from the conventional picture in which one QLL phase appear uniformly on ice crystal surfaces.

Cell‐friendly regulation of ice crystals by antifreeze organism‐inspired materials

AbstractThere is a dilemma between ice growth inhibition and cell‐threatening sharp‐edged ice formation, dominating an unfavorable efficiency of cell cryoprotection using ice control materials (ICMs). Herein, we design different‐type antifreeze organism‐inspired ICMs and show the cell‐friendly regulation of ice crystals by a synergistic effect for cell cryoprotection. Preferred rapid adsorption of “high‐diffusivity ICM” on the curved ice crystal surface induced by “ice‐binding ICM,” owing to the Gibbs–Curie–Wulff theorem, promotes smooth‐shaped and small ice crystal formation during the freeze–thaw cycle. Meanwhile, their intracellular and extracellular osmotic balance well protects the cryopreserved cells. All‐round protection benefits cell survival to achieve a cryopreservation efficiency of 97%. After cryopreservation, ice‐binding ICM exhibits double thermosensitive behaviors to directly encapsulate post‐thaw cells by gels, to further support cell‐based therapies. This work provides an in‐depth understanding of cell cryoprotection, and paves the road to the development of even better ICMs for biospecimen cryopreservation.

An insect antifreeze protein from Anatolica polita enhances the cryoprotection of Xenopus laevis eggs and embryos.

Cryoprotection is of interest in many fields of research, necessitating a greater understanding of different cryoprotective agents. Antifreeze proteins have been identified that have the ability to confer cryoprotection in certain organisms. Antifreeze proteins are an evolutionary adaptation that contributes to the freeze resistance of certain fish, insects, bacteria and plants. These proteins adsorb to an ice crystal's surface and restrict its growth within a certain temperature range. We investigated the ability of an antifreeze protein from the desert beetle Anatolica polita, ApAFP752, to confer cryoprotection in the frog Xenopus laevis. Xenopus laevis eggs and embryos microinjected with ApAFP752 exhibited reduced damage and increased survival after a freeze-thaw cycle in a concentration-dependent manner. We also demonstrate that ApAFP752 localizes to the plasma membrane in eggs and embryonic blastomeres and is not toxic for early development. These studies show the potential of an insect antifreeze protein to confer cryoprotection in amphibian eggs and embryos.