Micron-sized Crystals Research Articles

Endogenous Dynamic Nuclear Polarization for Natural Abundance 17O and Lithium NMR in the Bulk of Inorganic Solids.

In recent years magic angle spinning-dynamic nuclear polarization (MAS-DNP) has developed as an excellent approach for boosting the sensitivity of solid-state NMR (ssNMR) spectroscopy, thereby enabling the characterization of challenging systems in biology and chemistry. Most commonly, MAS-DNP is based on the use of nitroxide biradicals as polarizing agents. In materials science, since the use of nitroxides often limits the signal enhancement to the materials' surface and subsurface layers, there is need for hyperpolarization approaches which will provide sensitivity in the bulk of micron sized particles. Recently, an alternative in the form of paramagnetic metal ions has emerged. Here we demonstrate the remarkable efficacy of Mn(II) dopants, used as endogenous polarization agents for MAS-DNP, in enabling the detection of 17O at a natural abundance of only 0.038%. Distinct oxygen sites are identified in the bulk of micron-sized crystals, including battery anode materials Li4Ti5O12 (LTO) and Li2ZnTi3O8, as well as the phosphor materials NaCaPO4 and MgAl2O4, all doped with Mn(II) ions. Density functional theory calculations are used to assign the resonances to specific oxygen environments in these phases. Depending on the Mn(II) dopant concentration, we obtain significant signal enhancement factors, 142 and 24, for 6Li and 7Li nuclei in LTO, respectively. We furthermore follow the changes in the 6,7Li LTO resonances and determine their enhancement factors as a function of Mn(II) concentration. The results presented show that MAS-DNP from paramagnetic metal ion dopants provides an efficient approach for probing informative nuclei such as 17O, despite their low gyromagnetic ratio and negligible abundance, without isotope enrichment.

Micron-sized spinel crystals in high level waste glass compositions: Determination of crystal size and crystal fraction

The compositions utilized for immobilization of high-level nuclear wastes (HLW) are controlled using glass property models to avoid the deleterious effects of crystallization in the high-level waste (HLW) vitrification melters. The type and size of the crystals that precipitate during melter operations (typically at 1150 °C) and idling (∼1000 °C) are significantly impacted by glass composition and thermal history. This study was conducted to measure the impact of melt composition and heat treatment temperature on crystal size and fraction. A matrix of 31 multi-component glasses canvasing the expected Hanford HLW compositional space was developed and the glasses fabricated and heat treated at 850, 900, and 950 °C. The crystal amounts, as determined by X-ray diffraction, varied from 0.2 to 41.0 wt%. Spinel concentrations ranged from 0.0 to 13.8 wt%. One glass of the matrix did not precipitate spinel and contained 0.2 wt% RuO2, which was assumed to be undissolved from the melting process. All compositions contained crystals in the as-quenched glass. All of the spinel based crystals present in the glasses were less than 10 μm in diameter, as determined by scanning electron microscopy with image analysis. Composition and temperature dependent models were generated using the resulting data and the best model fit was obtained by only considering spinel concentrations (R2 = 0.87). Two glasses were unable to be characterized because of an inability to process the glass under the conditions of this study. Those glasses were utilized to give insight into a potential multi-component constraint to aid in future statistical composition designs.

Synthesis of Star-Shaped Boron Carbide Micro-Crystallites under High Pressure and High Temperatures

We synthesized star-shaped pentagonal microcrystals of boron carbide with an extremely low carbon content (~5%), from m-carborane under high pressure (7 GPa) and high temperature (1370–1670 K). These crystals have five-fold symmetry and grow in the shape of stars. A 5-fold symmetry in large micron-sized crystals is extremely rare making this a striking observation.

Improvement of activity and stability of Rhizomucor miehei lipase by immobilization on nanoporous aluminium oxide and potassium sulfate microcrystals and their applications in the synthesis of aroma esters

In this study, Rhizomucor miehei lipase (RML) was immobilized on the hexagonally-ordered nanoporous aluminium oxide membranes (RML-Al2O3-NP) by adsorption and as protein-coated microcrystals (RML-PCMCs) by simultaneously precipitating RML on micron-sized potassium sulfate crystals (K2SO4) in pre-chilled acetone. The hydrolytic activities of immobilized lipase preparations were investigated in terms of p-nitrophenyl palmitate hydrolysis and their esterification activities were examined for the synthesis of some aroma esters such as butyl acetate, isoamyl acetate, hexyl acetate, heptyl acetate, and geranyl acetate. The immobilization yields were 33.8 and 25.1%, respectively for RML immobilized on Al2O3-NP membranes and potassium sulfate crystals. The catalytic efficiency ratios of RML-Al2O3-NP and RML-PCMCs were 2.3- and 3.9-fold higher than that of the free lipase, respectively in terms of hydrolytic activity. The free lipase was stabilized as 4.1- and 10.5-fold, respectively at 40 and 50 °C when immobilized on Al2O3-NP. The corresponding stabilization factors were 4.6- and 12.8-fold higher for RML-PCMCs. RML-Al2O3-NP and RML-PCMCs maintained 84 and 86% of their initial hydrolytic activities, respectively after 10 reuses. Of the synthesized aroma esters, the highest yield was obtained for the geranyl acetate. After 4 h reaction time, no geraniol was detected in the preparative-scale (196 g/L) synthesis of geranyl acetate for both the immobilized lipases when the initial geraniol amount, vinyl acetate amount, RML-PCMCs amount, and reaction temperature values were 1 mmol, 3 mmol, 100 mg (or 300 mg RML-Al2O3-NP), and 50 °C, respectively. These results show that the immobilization of R. miehei lipase by adsorption on nanoporous aluminium oxide and as protein-coated microcrystals leads to the obtention of highly stable, catalytically more active, and reusable lipase preparations.

The "speed limit" for macromolecular crystal growth.

A simple “diffusion‐to‐capture” model is used to estimate the upper limit to the growth rate of macromolecular crystals under conditions when the rate limiting process is the mass transfer of sample from solution to the crystal. Under diffusion‐limited crystal growth conditions, this model predicts that the cross‐sectional area of a crystal will increase linearly with time; this prediction is validated by monitoring the growth rate of lysozyme crystals. A consequence of this analysis is that when crystal growth is diffusion‐limited, micron‐sized crystals can be produced in ~1 s, which would be compatible with the turnover time of many enzymes. Consequently, the ability to record diffraction patterns from sub‐micron sized crystals by X‐ray Free Electron Lasers and micro‐electron diffraction technologies opens the possibility of trapping intermediate enzyme states by crystallization.

Potassium- and Rubidium-Passivated Alloyed Perovskite Films: Optoelectronic Properties and Moisture Stability.

Halide perovskites passivated with potassium or rubidium show superior photovoltaic device performance compared to unpassivated samples. However, it is unclear which passivation route is more effective for film stability. Here, we directly compare the optoelectronic properties and stability of thin films when passivating triple-cation perovskite films with potassium or rubidium species. The optoelectronic and chemical studies reveal that the alloyed perovskites are tolerant toward higher loadings of potassium than rubidium. Whereas potassium complexes with bromide from the perovskite precursor solution to form thin surface passivation layers, rubidium additives favor the formation of phase-segregated micron-sized rubidium halide crystals. This tolerance to higher loadings of potassium allows us to achieve superior luminescent properties with potassium passivation. We also find that exposure to a humid atmosphere drives phase segregation and grain coalescence for all compositions, with the rubidium-passivated sample showing the highest sensitivity to nonperovskite phase formation. Our work highlights the benefits but also the limitations of these passivation approaches in maximizing both optoelectronic properties and the stability of perovskite films.

Zeolite-like MOF nanocrystals incorporated 6FDA-polyimide mixed-matrix membranes for CO2/CH4 separation

MOF mixed-matrix membranes (MMMs) are regarded as promising candidates for energy-efficient natural gas purification. This work reports the fabrication of high-performance 6FDA-polyimide MMMs, based on the incorporation of zeolite-like MOF (ZMOF) as fillers, and their associated permeation studies for CO2/CH4 separation. To eliminate micron-sized crystals, a facile repeating sedimentation approach was used to harvest nanocrystals from the as-synthesized bulk ZMOF crystalline powder material. This enables the deployment of ZMOF nanocrystals with relatively uniform dimension and morphology in the polymer matrix. Typical 6FDA-polyimides encompassing distinct diamine moieties (6FDA-DAM, 6FDA-DETDA-DABA or PDMC) were explored as polymer matrices to disclose the transport property matching the hosted ZMOF filler. Mixed-gas permeation measurements revealed that the incorporation of the ZMOF filler affords a concurrent enhancement of the CO2 permeability and the CO2/CH4 selectivity for the three tested 6FDA-polyimides. Particularly, the highly permeable 6FDA-DAM showed a considerably enhanced performance for CO2/CH4 that transcends the 2008 Robeson upper-bound. Detailed analysis of the sorption data and diffusion coefficients suggest that the enhanced transport property in the ZMOF-based MMM is plausibly attributed to the combination of the higher CO2 sorption capacity and selectivity, and favorable gas diffusivity via the CO2-philic framework of ZMOF in moderately confined pores.

Instability of a Noncrystalline NaO2 Film in Na–O2 Batteries: The Controversial Effect of the RuO2 Catalyst

The unique electrochemical and chemical features of sodium–oxygen (Na–O2) batteries distinguish them from the lithium–oxygen (Li–O2) batteries. NaO2 as the main discharge product is unstable in the cell environment and chemically degrades, which triggers side products’ formation and charging potential increment. In this study, RuO2 nanoparticles dispersed on carbon nanotubes (CNTs) are used as the catalyst for Na–O2 batteries to elucidate the effect of the catalyst on these complex electrochemical systems. The RuO2/CNT contributes to the formation of a poorly crystalline and coating-like NaO2 structure during oxygen reduction reaction, which is drastically different from the conventional micron-sized cubic NaO2 crystals deposited on the CNT. Our findings demonstrate a competition between NaO2 and side products’ decompositions for RuO2/CNT during oxygen evolution reaction (OER). We believe that this is due to the lower stability of a coating-like NaO2 because of its noncrystalline nature and high electrode...

Photophysical properties of cationic dyes captured in the mesoscale channels of micron-sized metal-organic framework crystals

The optical properties of dye molecules in confined spaces can differ from the solution phase due to confinement effects. Pre-organized mesoscale channels of metal-organic frameworks (MOFs) are very suited for hosting various dyes, and the robust frameworks often render the encapsulated dyes with certain preferential geometries, which are different from those found in solution. Furthermore, pre-organized open channels can efficiently guide the uniform and unique spatial distribution of dye molecules in a controlled manner, which are otherwise difficult to achieve. Thus, sufficiently large dye molecules can avoid the formation of complex aggregates when captured inside open channels. In contrast, small dye molecules can form well-defined dimers or aggregates. The resulting dye-encapsulated MOFs can display unusual photophysical properties of the captured dyes. An anionic framework of In-BTB with mesoscale 3D channels is utilized for the efficient encapsulation of various cationic dyes through cation-exchange processes. Six different cationic dyes are encapsulated in the anionic framework of In-BTB, and their crystal structures are completely solved. Novel photophysical properties of these spatially distributed dye molecules in dye@In-BTBs are investigated.

The Influence of Photoelectron Escape in Radiation Damage Simulations of Protein Micro-Crystallography

Radiation damage represents a fundamental limit in the determination of protein structures via macromolecular crystallography (MX) at third-generation synchrotron sources. Over the past decade, improvements in both source and detector technology have led to MX experiments being performed with smaller and smaller crystals (on the order of a few microns), often using microfocus beams. Under these conditions, photoelectrons (PEs), the primary agents of radiation-damage in MX, may escape the diffraction volume prior to depositing all of their energy. The impact of PE escape is more significant at higher beam energies (>20 keV) as the electron inelastic mean free path (IMFP) is longer, allowing the electrons to deposit their energy over a larger area, extending further from their point of origin. Software such as RADDOSE-3D has been used extensively to predict the dose (energy absorbed per unit mass) that a crystal will absorb under a given set of experimental parameters and is an important component in planning a successful MX experiment. At the time this study was undertaken, dose predictions made using RADDOSE-3D were spatially-resolved, but did not yet account for the propagation of PEs through the diffraction volume. Hence, in the case of microfocus crystallography, it is anticipated that deviations may occur between the predicted and actual dose absorbed due to the influence of PEs. To explore this effect, we conducted a series of simulations of the dose absorbed by micron-sized crystals during microfocus MX experiments. Our simulations spanned beam and crystal sizes ranging from 1μm to 5μm for beam energies between 9 keV and 30 keV. Our simulations were spatially and temporarily resolved and accounted for the escape of PEs from the diffraction volume. The spatially-resolved dose maps produced by these simulations were used to predict the rate of intensity loss in a Bragg spot, a key metric for tracking global radiation damage. Our results were compared to predictions obtained using a recent version of RADDOSE-3D that did not account for PE escape; the predicted crystal lifetimes are shown to differ significantly for the smallest crystals and for high-energy beams, when PE escape is included in the simulations.

Dislocation-Free SiGe/Si Heterostructures

Ge vertical heterostructures grown on deeply-patterned Si(001) were first obtained in 2012 (C.V. Falub et al., Science2012, 335, 1330–1334), immediately capturing attention due to the appealing possibility of growing micron-sized Ge crystals largely free of thermal stress and hosting dislocations only in a small fraction of their volume. Since then, considerable progress has been made in terms of extending the technique to several other systems, and of developing further strategies to lower the dislocation density. In this review, we shall mainly focus on the latter aspect, discussing in detail 100% dislocation-free, micron-sized vertical heterostructures obtained by exploiting compositional grading in the epitaxial crystals. Furthermore, we shall also analyze the role played by the shape of the pre-patterned substrate in directly influencing the dislocation distribution.

Photoluminescence Flickering of Micron-Sized Crystals of Methylammonium Lead Bromide: Effect of Ambience and Light Exposure

Recent reports on temporal photoluminescence (PL) intensity fluctuations (blinking) within localized domains of organo-metal lead halide (hybrid) perovskite microcrystals have invoked considerable interest to understand their origins. Using PL microscopy, we have investigated the effect of atmospheric constituents and photoillumination on spatially extended intensity fluctuations in methylammonium lead bromide (MAPbBr3) perovskite materials, explicitly for micrometer (ca. 1–2 μm)-sized crystals. Increase in the relative humidity of the ambience results in progressive reduction in the PL intensity, and beyond a threshold value, individual microcrystalline grains exhibit multistate PL intermittency (flickering), which is characteristically different from quasi two-state blinking observed in nanocrystals. Such flickering disappears upon removal of moisture, accompanied by considerable enhancement of the overall PL efficiency. We hypothesize that initiation of moisture-induced degradation marked by the loweri...

Quick, Easy, and Economic Mineralogical Studies of Flooded Chalk for EOR Experiments Using Raman Spectroscopy

Understanding the chalk-fluid interactions and the associated mineralogical and mechanical alterations on a sub-micron scale are major goals in Enhanced Oil Recovery. Mechanical strength, porosity, and permeability of chalk are linked to mineral dissolution that occurs during brine injections, and affect the reservoir potential. This paper presents a novel “single grain” methodology to recognize the varieties of carbonates in rocks and loose sediments: Raman spectroscopy is a non-destructive, quick, and user-friendly technique representing a powerful tool to identify minerals down to 1 µm. An innovative working technique for oil exploration is proposed, as the mineralogy of micron-sized crystals grown in two flooded chalk samples (Liége, Belgium) was successfully investigated by Raman spectroscopy. The drilled chalk cores were flooded with MgCl2 for ca. 1.5 (Long Term Test) and 3 years (Ultra Long Term Test) under North Sea reservoir conditions (Long Term Test: 130 °C, 1 PV/day, 9.3 MPa effective stress; Ultra Long Term Test: 130 °C, varying between 1–3 PV/day, 10.4 MPa effective stress). Raman spectroscopy was able to identify the presence of recrystallized magnesite along the core of the Long Term Test up to 4 cm from the injection surface, down to the crystal size of 1–2 µm. In the Ultra Long Term Test core, the growth of MgCO3 affected nearly the entire core (7 cm). In both samples, no dolomite or high-magnesium calcite secondary growth could be detected when analysing 557 and 90 Raman spectra on the Long and Ultra Long Term Test, respectively. This study can offer Raman spectroscopy as a breakthrough tool in petroleum exploration of unconventional reservoirs, due to its quickness, spatial resolution, and non-destructive acquisition of data. These characteristics would encourage its use coupled with electron microscopes and energy dispersive systems or even electron microprobe studies.

Supersaturation-controlled microcrystallization and visualization analysis for serial femtosecond crystallography

Time-resolved serial femtosecond crystallography with X-ray free electron laser (XFEL) holds the potential to view fast reactions occurring at near-physiological temperature. However, production and characterization of homogeneous micron-sized protein crystals at high density remain a bottleneck, due to the lack of the necessary equipments in ordinary laboratories. We describe here supersaturation-controlled microcrystallization and visualization and analysis tools that can be easily used in any laboratory. The microcrystallization conditions of the influenza virus hemagglutinin were initially obtained with low reproducibility, which was improved by employing a rapid evaporation of hanging drops. Supersaturation-controlled microcrystallization was then developed in a vapor diffusion mode, where supersaturation was induced by evaporation in hanging drops sequentially for durations ranging from 30 sec to 3 min, depending on the protein. It was applied successfully to the microcrystal formation of lysozyme, ferritin and hemagglutinin with high density. Moreover, visualization and analysis tools were developed to characterize the microcrystals observed by light microscopy. The size and density distributions of microcrystals analyzed by the tools were found to be consistent with the results of manual analysis, further validated by high-resolution microscopic analyses. Our supersaturation-controlled microcrystallization and visualization and analysis tools will provide universal access to successful XFEL studies.

NE-CAT: Crystallography Beamlines for Challenging Structural Biology Research

The NorthEastern Collaborative Access Team (NE-CAT) focuses on the design and operation of synchrotron X-ray beamlines for the solution of technically challenging structural biology problems and provides an important resource for the national and international research community. Currently, NE-CAT operates two undulator beamlines: a 6 - 22 keV tunable energy beamline (24-ID-C) and a 12.662 keV single energy beamline (24-ID-E). Both beamlines are equipped with state-of-the-art instrumentation. MD2 microdiffractometers installed at both beamlines provide very clean beams down to 5 microns in diameter and are capable of visualizing micron-sized crystals. Large area pixel array detectors (Pilatus6M and Eiger16M) provide fast noiseless data collection and make possible it to resolve large unit cells. Both beamlines are equipped with custom-built ALS-style robotic sample automounters with dewars capable of holding 14 pucks. Our locally developed software suite RAPD provides data collection strategies, quasi-real time data integration, scaling and simple automated MR/SAD pipeline through a 384-core compute cluster. Towards improving the diffraction from low resolution crystals, NE-CAT has installed humidity controlled device, HC1 in the 24ID-E hutch. Users of the beamlines are supported 24/7 by experienced resident crystallographers. Funding for NE-CAT is provided through a P41 grant from the NIGMS and from the NE-CAT member institutions.

Listwaenite in the Sartohay ophiolitic mélange (Xinjiang, China): A genetic model based on petrology, U-Pb chronology and trace element geochemistry

Listwaenite lenses in the Sartohay ophiolitic mélange (Xinjiang, China) were formed via reactions between serpentinite and metasomatic fluids. First, serpentinite changed into talc schist via the reaction of serpentine + CO2 → talc + magnesite + H2O. Second, talc schist changed into listwaenite via the reaction of talc + CO2 → magnesite + quartz + H2O. Magnetite was progressively destroyed during transformation from serpentinite to talc schist, and completely consumed in listwaenite. Zircon crystals 30–100 μm long, disseminating in talc schist, undeformed listwaenite and mylonitized listwaenite, coexist with talc, quartz and magnesite, while micron-sized zircon grains (<5 μm in length) occur along the shearing foliation in the weakly deformed listwaenite and mylonitized listwaenite. We postulate that these micron-sized zircon crystals may have grown in-situ from medium-temperature hydrothermal fluids. Concentrations of most trace elements including high field strength elements (HFSE) increase from the undeformed, through the weakly deformed, to the mylonitized listwaenite, showing a positive correlation with the degrees of deformation and proportions of micron-size zircon, apatite, rutile and monazite. The large zircon crystals recovered from talc schist, undeformed listwaenite and mylonitized listwaenite yield similar weighted mean U-Pb ages (302.9 ± 6.8 Ma, 299.7 ± 5.5 Ma and 296.5 ± 3.5 Ma), and are thought to represent the age of formation of the talc schist and listwaenite. These ages are indistinguishable within errors and suggest a rapid transformation from talc schist to listwaenite. Some zircon rims in samples of the undeformed listwaenite and mylonitized listwaenite give much younger apparent U-Pb ages (280–277 Ma), which could be interpreted as a recrystallization age reflecting late-stage shearing in the Sartohay ophiolitic mélange.

AFM Nanoindentation To Quantify Mechanical Properties of Nano- and Micron-Sized Crystals of a Metal-Organic Framework Material.

The mechanical properties of individual nanocrystals and small micron-sized single crystals of metal-organic frameworks (MOFs), hitherto, cannot be measured directly by employing the conventional instrumented nanoindentation approach. Here we propose the application of atomic force microscopy (AFM)-based nanoindentation technique, equipped with a calibrated diamond cube-corner indenter tip to quantify the Young's modulus, hardness, adhesion energy, and interfacial and fracture strengths of a zeolitic imidazolate framework (ZIF-8) porous material. We use ZIF-8 as a model MOF system to develop AFM nanoindentation leveraging the concept of unloading strain rate, enabling us to critically assess the practicality and technical limitations of AFM to achieve quantitative measurements of fine-scale MOF crystals. We demonstrate the advantages of using a high unloading strain rate (ε̇ > 60 s-1) to yield reliable force-displacement data in the few μN load range, corresponding to a shallow indentation depth of ∼10s nm. We found that the Young's moduli (∼3-4 GPa) determined by AFM nanoindentation of the nanocrystals (<500 nm) and micron-sized crystals (∼2 μm) are in agreement with magnitudes derived previously from other techniques, namely instrumented nanoindentation and Brillouin spectroscopy (however, these methods requiring large 100-μm sized crystals) and also in line with density functional theory predictions of an idealized ZIF-8 crystal.

Size and Shape Control of Micron-Sized Salicylic Acid Crystals during Antisolvent Crystallization

During an antisolvent crystallization process, micron-sized crystals of salicylic acid in the particle size range 20 to 150 μm were prepared under specific sets of crystallization conditions, with a focus on drug concentration, temperature, and solvent composition. For each experiment, the size outcome was determined by a number of knock on (often competing) influencing factors, including supersaturation, mass available for deposition during crystallization, and influence of temperature and solvent composition on crystallization kinetics. A certain fraction of the crystals, especially at higher solute concentration, developed a spectacular hollow shape with an almost perfect rectangular outer cross-section. A mechanistic explanation for the formation of hollow crystals is proposed. The use of additives during crystallization introduced further control over the size and, perhaps more noticeably, over the shape of the salicylic acid crystals. The lengths of salicylic acid crystals decreased to as low as 6 μ...

Chemical aspect of the structural disorder in CuCrS2 and CuCr1–x V x S2 solid solutions

A gradient chemical structure of micron-sized crystals of CuCrS2 and CuCr1–x V x S2 powders is established by the differential dissolution technique. It is shown that their surface region coherently conjugated with planar faces of CuCrS2 crystals is enriched with copper whereas the bulk is depleted of copper and the copper sublattice is disordered. A possible mechanism of the formation of surface regions due to a high bulk mobility of copper atoms in the defect copper sublattice is proposed. It is shown that for solid solutions the disordering process involves copper and vanadium atoms and vacancy complexes. The phenomena found are discussed together with the structural and magnetic data.

Time-dependent subcellular structure injuries induced by nano-/micron-sized calcium oxalate monohydrate and dihydrate crystals

Comparative studies were conducted to investigate the time effect of cell injury induced by nano-sized (50nm) and micron-sized (10μm) calcium oxalate monohydrate (COM) and dihydrate (COD) crystals in African green monkey renal epithelial (Vero) cells. The effects of nano-/micron-sized COM and COD exposure on Vero cells were investigated by detecting the cell viability, cell morphology, LDH release, reactive oxygen species, mitochondrial membrane potential, cell cycle, and cell apoptosis, as well as the intracellular and extracellular crystal distribution. Nano-/micron-sized COM and COD exposure lead to subcellular organelle injury in varying degrees, but the injury sequence of various organelles differed. The time sequence of organelle injury presenting significant variation was described as follows: cell membrane injury (1h)<mitochondrial membrane potential decrease (3h to 6h)≈cell-cycle arrest (3h to 6h)<cell apoptosis (12h). Nano-sized crystals lead organelle injury faster than micron-sized crystals, and COM crystals showed more obvious time-dependent effects than the same-sized COD crystals. This study may provide insights into the damage to renal epithelial cells induced by urinary crystals and the formation mechanism of kidney stones.