Polarized Light Microscopy Research Articles

In vitro caries-preventive effect of a mineralization-promoting peptide combined with fluoride gel on sound primary teeth.

Mineralization-promoting peptide-3 (MPP3) is a new biomimetic remineralization agent. To assess the remineralization efficiency of MPP3, either alone or in combination with fluoride gel. The samples were divided into four groups: control, 1.23% fluoride gel, 10% MPP3 gel, and 1.23% fluoride gel + 10% MPP3. Following the application of remineralization agents (4 min), the samples remained in a pH-cycling model (37°C, 4 weeks). Microhardness, microcomputed tomography (micro-CT), polarized light microscopy (PLM), and field emission scanning electron microscopy (FE-SEM) analysis were conducted. RM-ANOVA, one-way ANOVA, and intraclass correlation coefficient (ICC) were used for statistical analysis, and a significance level of p < .05 was employed. Mineralization-promoting peptide 3 and fluoride gel + MPP3 increased the microhardness of the enamel compared with initial values in each group (p < .05). Mineralization-promoting peptide 3 successfully maintained the mineral density of enamel, although the cariogenic pH-cycling and PLM results indicated that the lesion depth (μm) was significantly lower in the fluoride gel + MPP3 group (27.0336 ± 12.53650) than in the control group (37.3907 ± 12.76002, p < .05). The combined use of MPP3 with fluoride gel enhanced the caries-protective and mineralization-promoting effects of fluoride. Mineralization-promoting peptide 3 may be a potential agent that can be employed to improve the physical properties of enamel.

The frieze of the Art of War in the Ducal Palace of Urbino: conservation issues, materials, and executive techniques

The frieze of the Art of War was originally positioned along the sides of the facade of the Ducal Palace in Urbino (Marche, Italy), and it is currently located in the palace’s Chancellery Room. The frieze, probably elaborated by Federico da Montefeltro around 1474, consists s of stone bas-reliefs illustrating a rich iconographic repertoire depicting numerous engineering machines and symbols related to the military and political spheres. The present work aims to deepen the knowledge of the Art War frieze and to clarify some doubts regarding the original shape of the bas-reliefs, the constituent material, and the executive technique; conservation issues were also considered. Polarized light microscopy observations were carried out on the stone materials, identifying the use of a packstone referable to the Calcare Massiccio A Formation (Hettangian-Lower Pliensbachian p.p.). This stone, locally known as Piobbico Travertine, was excavated in the Val d’Abisso quarries, part of the Umbria-Marche ridge. Ca-oxalates were detected by Fourier transform infrared analysis in the brownish patinas that mainly cover the surfaces of the bas-reliefs; they could be related to ancient conservation treatments and/or to biological agents. Regarding the execution technique, a thorough autoptic examination allowed for a better understanding of the original conformation of the bas-reliefs and the carving process. Stylistic and executive differences in the execution of the same detail were observed in different bas-reliefs, confirming the activity of various sculptors. Different decay typologies were used to consider the debate regarding the original arrangement of the frieze on the facade, validating the most accredited hypothesis.

Role of Stearic Acid as the Crystal Habit Modifier in Candelilla Wax-Groundnut Oil Oleogels

This study investigated the effects of incorporating stearic acid (SAC) in candelilla wax (CW) and groundnut oil (GO) oleogel with potential health benefits as an alternative to saturated fats in processed foods. Results showed that SAC possesses crystal habit-modifying properties on the oleogels, causing its average crystallite size to increase, as observed through polarized light microscopy and XRD analysis. Additionally, SAC caused an increase in ordering within the crystallite network as a result of the decrease in d-spacing. Interestingly, the firmness of the oleogels remained unaffected, even at a higher fraction of SAC. It is believed to be due to the interference caused by the crystallization of high-melting SAC within the fine crystal network of CW-GO oleogel. However, adding 3 mg of SAC significantly increased the work of the shear of the oleogel (SAC3), which decreased the spreadability. As observed through colorimetric analysis, SAC3 showed a dense and uniform distribution of prominent bright crystals with minimal amorphous regions, leading to a high whiteness index. SAC3 also demonstrated the highest compactness and dislocation density among the oleogels, likely due to the formation of prominent crystals. However, SAC did not affect the overall oleogel crystallization rate. SAC3 had delayed secondary crystallization and thermal equilibrium by having a prolonged crystallization time of CW crystals. In the case of controlled delivery studies, the addition of SAC improved CPCR. On the other hand, CPCR decreased with the increase in SAC amount, where SAC3 showed a moderate curcumin release ability among the oleogels.

Enhancement of flexural properties of carbon fiber-reinforced polyamide 6 via oriented crystallization of polyamide 6 among carbon fibers

The effect of the crystalline structure of polyamide 6 (PA6) among carbon fiber (CF) on the mechanical properties of PA6/CF composites was explored using two types of PA6 with different spherulite sizes. Infrared (IR) spectroscopy showed that intra/intermolecular interactions were nearly equivalent for the two types of PA6. The viscoelasticities in the molten state were almost identical; that is, the molecular weights of the PA6s were comparable. The mechanical properties of the PA6 matrix with smaller spherulites (PA6a) in PA6/CF were superior to those of the PA6 matrix with larger spherulites (PA6b). Polarized light microscopy, wide-angle X-ray scattering, and polarized Raman spectroscopy revealed oriented crystals of PA6a in the PA6a/CF composite and random crystals of PA6b in the PA6b/CF composite. Therefore, the superior mechanical properties of PA6/CF are due to the oriented crystal chains of PA6, which support the overall stress transfer in composites.

Mechanical Stress Decreases the Amplitude of Twisting and Bending Fluctuations of Actin Filaments

A variety of biological roles of mechanical forces have been proposed in cell biology, such as cell signaling pathways for survival, development, growth, and differentiation. Mechanical forces alter the mechanical conditions within cells and their environment, which strongly influences the reorganization of the actin cytoskeleton. Single-molecule imaging studies of actin filaments have led to the hypothesis that the actin filament acts as a mechanosensor; e.g., increases in actin filament tension alter their conformation and affinity for regulatory proteins. However, our understanding of the molecular mechanisms underlying how tension modulates the mechanical behavior of a single actin filament is still incomplete. In this study, a direct measurement of the twisting and bending of a fluorescently labeled single actin filament under different tension levels by force application (0.8–3.4 pN) was performed using single-molecule fluorescence polarization (SMFP) microscopy. The results showed that the amplitude of twisting and bending fluctuations of a single actin filament decreased with increasing tension. Electron micrograph analysis of tensed filaments also revealed that the fluctuations in the crossover length of actin filaments decreased with increasing filament tension. Possible molecular mechanisms underlying these results involving the binding of actin-binding proteins, such as cofilin, to the filament are discussed.

The flow of anisotropic nanoparticles in solution and in blood.

The alignment of anisotropic nanoparticles in flow has been used for a range of applications such as the preparation of strong fibres and the assembly of in-plane aligned 1D-nanoobjects that are used for electronic devices, sensors, energy and biological application. Important is also the flow behaviour of nanoparticles that were designed for nanomedical applications such as drug delivery. It is widely observed that non-spherical nanoparticles have longer circulation times and a more favourable biodistribution. To be able to understand this behaviour, researchers have turned to analyzing the flow of non-spherical nanoparticles in the blood stream. In this review, an overview of microfluidic techniques that are used to monitor the alignment of anisotropic nanoparticles in solution will be provided, which includes analysis by small angle X-ray scattering (SAXS) and polarized light microscopy. The flow of these nanoparticles in blood is then discussed as the presence of red blood cells causes margination of some nanoparticles. Using fluorescence microscopy, the extent of margination can be identified, which coincides with the ability of nanoparticles to adhere to the cells grown along the wall. While these studies are mainly carried out in vitro using blood, initial investigations in vivo were able to confirm the unusual flow of anisotropic nanoparticles.

Birefringence after Tellurium Nanosheets and Copper Intercalation

Symmetry-deficient two-dimensional (2D) layered materials induce a highly anisotropic optical response due to the anisotropy in their crystal structure, facilitating their application in polarized nanodevices. Intercalation is a new way to tune the optoelectronic properties of materials by inserting guest atoms into layered host materials, and 2D layered structures stacked with van der Waals gaps are a prerequisite for this phase of the technique. In this paper, 2D tellurium nanosheets were synthesized with a hydrothermal method, and copper atoms were inserted with a wet chemical method. The widening of the crystal plane spacing proves the introduction of copper atoms, and polarization−related second-harmonic-generation (SHG) studies reveal the intrinsic anisotropic modes of the two samples, and birefringent properties are found with polarizing light microscopy. We further investigated the electrical properties of the samples, and the embedding of the copper atoms caused the samples to exhibit higher currents, but their devices lost the gate control effect.

Mesostructured Fibrils Exfoliated in Deep Eutectic Solvent as Building Blocks of Collagen Membranes.

The mesoscale components of collagen (nanofibrils, fibrils, and fiber bundles) are well organized in native tissues, resulting in superior properties and diverse functions. In this paper, we present a simple and controlled liquid exfoliation method to directly extract medium-sized collagen fibers ranging from 102 to 159 nm in diameter from bovine Achilles tendon using urea/hydrochloric acid and a deep eutectic solvent (DES). In situ observations under polarized light microscopy (POM) and molecular dynamics simulations revealed the effects of urea and GuHCl on tendon collagen. FTIR study results confirmed that these fibrils retained the typical structural characteristics of type I collagen. These shed collagen fibrils were then used as building blocks to create independent collagen membranes with good and stable mechanical properties, excellent barrier properties, and cell compatibility. A new method for collagen processing is provided in this work by using DES-assisted liquid exfoliation for constructing robust collagen membranes with mesoscale collagen fibrils as building blocks.

Study of the physico-chemical properties of oligoephiramides based on 4-aminobenzoic acid

This work was carried out to obtain a number of aromatic oligoetheramides and to evaluate the effect of their structure on mesomorphic and thermal properties. Oligoetheramides based on 4-aminobenzoic acid (4-ABA) were synthesized by hightemperature polycondensation. The structure of the obtained compounds was identified by IR spectroscopy, the thermal parameters were determined from the data of thermogravimetric analysis (TGA), the phase transition temperatures were determined by differential scanning calorimetry (DSC), and the manifestation of the optical activity of oligoetheramide solutions was studied by optical polarization microscopy (OPM). It was revealed that the samples dissolve when heated in sulfuric acid, in dimethylformamide (DMF), dimethylacetamide (DMAA) and have limited solubility in dimethylsulfoxide (DMSO). Solutions of oligomers exhibited optical activity at crossed microscope polarizers.

Carrageenan tethered ion sensitive smart nanogel containing oleophytocubosomes for improved ocular luteolin delivery

Ophthalmic delivery of luteolin (LU) was studied after formulating a carrageenan-based novel ion-sensitive in situ gel (ISG) incorporating oleophytocubosomes for prolonged ocular residence time and improved ocular bioavailability of the poorly absorbed herbal drug luteolin. The prepared oleophytocubosomes and ISG were compared with LU suspension. Optimized oleophytocubosomes possessed small, homogenously distributed negatively charged particles with high entrapment efficiency. Polarized light microscope revealed a cubic phase. Optimized ISG matrix composed of 0.4% kappa carrageenan (KC), and 2% hydroxypropylmethylcellulose (HPMC) demonstrated rapid gelation, high resistance to dilution, increased viscosity after gelation, and strong mucoadhesive properties. oleophytocubosomes exerted improved drug release, while a more sustained release was observed for ISG oleophytocubosomes. The antioxidant activity of both formulations was significantly higher than that of LU suspension. Oleophytocubosome and ISG oleophytocubosome revealed significantly higher apparent permeability coefficients of 3.62 and 2.90 folds, respectively, compared to LU suspension. Irritation tests showed the safety of both formulations for single- and multiple-ocular administration. In-vivo studies demonstrated that the ISG system showed prolonged antiglaucoma effects and a faster anti-inflammatory effect, followed by oleophytocubosomes.

Solvent and Crystallization Effects on the Dermal Absorption of Hydrophilic and Lipophilic Compounds

This study probes the mechanisms by which volatile solvents (water, ethanol) and a nonionic surfactant (Triton X-100) influence the skin permeation of dissolved solutes following deposition of small doses onto unoccluded human skin. A secondary objective was to sharpen guidelines for the use of these and other simple solvent systems for dermal safety testing of cosmetic ingredients at finite doses. Four solutes were studied – niacinamide, caffeine, testosterone and geraniol – at doses close to that estimated to saturate the upper layers of the stratum corneum. Methods included tensiometry, visualization of spreading on skin, polarized light microscopy and in vitro permeation testing using radiolabeled solutes. Ethanol, aqueous ethanol and dilute aqueous Triton solutions all yielded surface tensions below 36 mN/m, allowing them to spread easily on the skin, unlike water (72.4 mN/m) which did not spread. Deposition onto skin of niacinamide (32 μg·cm−2) or caffeine (3.2 μg·cm−2) from water and ethanol led to crystalline deposits on the skin surface, whereas the same amounts applied from aqueous ethanol and 2 % Triton did not. Skin permeation of these compounds was inversely correlated to the extent of crystallization. A separate study with caffeine showed the absence of a dose-related skin permeability increase with Triton. Permeation of testosterone (8.2 μg·cm−2) was modestly increased when dosed from aqueous ethanol versus ethanol. Permeation of geraniol (2.9 μg·cm−2) followed the order aqueous ethanol > water ∼ 2 % Triton >> ethanol and was inversely correlated with evaporative loss. We conclude that, under the conditions tested, aqueous ethanol and Triton serve primarily as deposition aids and do not substantially disrupt stratum corneum lipids. Implications for the design of in vitro skin permeability tests are discussed.

Thermomechanical Properties of High Oleic Palm Oil Assessed Using Differential Scanning Calorimetry, Texture Analysis, Microscopy, and Shear Rheology.

Standard Palm Oil (SPO) is widely used as a food ingredient partially due to its unique thermophysical properties. However, the American Heart Association recommends a saturated fat consumption of <5% of the caloric intake per day. The OxG Palm hybrid yields oil known as "palm oil with a higher content of oleic acid" (HOPO), with <35% SFA and >50% oleic acid. Characterizing novel high oleic oils is the starting point to find processes that can functionalize them such as oleogelation. This study compared the thermophysical properties of HOPO to SPO using Differential Scanning Calorimetry, shear rheology, polarized light microscopy, and texture analysis to characterize the differences between these oils. HOPO had a lower onset crystallization temperature (Δ7 °C) and its rheological behavior followed similar trends to SPO; however, large viscosity offsets were observed and were correlated to differences in crystallization temperatures. The maximum peak force of SPO was an order of magnitude higher than that of HOPO. Overall similar trends between the oils were observed, but differences in firmness, crystal morphology, and viscosity were not linearly correlated with the offset in crystallization temperature. This study quantified differences between these oils that will better enable industry to use HOPO in specific applications.

Detection of Methicillin-Resistant Staphylococcus Aureus Bacteria Using Liquid Crystals

Methicillin-resistant Staphylococcus aureus (MRSA) is an essential pathogen for public health and this bacteria commonly cause serious infectious in humans. In recent years, MRSA bacteria are detected by the bacterial culture and nucleic acid-based methods which are time-consuming and labor-intensive. In this study, a novel liquid crystal (LC)-based biosensing system was developed to overcome these limitations. The objective of this study was to detect the presence of MRSA bacteria which prepared within the isotonic water and phosphate buffer saline (PBS). In this system, the binding of MRSA bacteria to the dimethyloctadecyl [3-(trimethoxysilyl) propyl] ammonium chloride (DMOAP)-coated surface disrupted the orientation of LCs, triggering a transition from a homeotropic orientation to a random one. This transition in the orientation of the LCs was observed as a change from a dark optical LC image to a bright one under a polarized light microscope (POM) and the reflection values of LC molecules were determined by using a spectrometer. Through this sensing mechanism, MRSA bacteria prepared within the isotonic water was detected ranging from the 9.2x103 CFU/mL to 9.2x107 CFU/mL concentration. Furthermore, MRSA bacteria prepared in PBS was detected in the concentration range of 7.1x104 CFU/mL to 7.1x108 CFU/mL by using this system.

Influence of hematoxylin and eosin staining on linear birefringence measurement of fibrous tissue structures in polarization microscopy.

For microscopic polarization imaging of tissue slices, two types of samples are often prepared: one unstained tissue section for polarization imaging to avoid possible influence from staining dyes quantitatively and one hematoxylin-eosin (H&E) stained adjacent tissue section for histological diagnosis and structural feature identification. However, this sample preparation strategy requires high-quality adjacent tissue sections, and labeling the structural features on unstained tissue sections is impossible. With the fast development of data driven-based polarimetric analysis, which requires a large amount of pixel labeled images, a possible method is to directly use H&E stained slices, which are standard samples archived in clinical hospitals for polarization measurement. We aim to study the influence of hematoxylin and eosin staining on the linear birefringence measurement of fibrous tissue structures. We examine the linear birefringence properties of four pieces of adjacent bone tissue slices with abundant collagen fibers that are unstained, H&E stained, hematoxylin (H) stained, and eosin (E) stained. After obtaining the spatial maps of linear retardance values for the four tissue samples, we carry out a comparative study using a frequency distribution histogram and similarity analysis based on the Bhattacharyya coefficient to investigate how H&E staining affects the linear birefringence measurement of bone tissues. Linear retardance increased after H&E, H, or E staining (41.7%, 40.8%, and 72.5% increase, respectively). However, there is no significant change in the imaging contrast of linear retardance in bone tissues. The linear retardance values induced by birefringent collagen fibers can be enhanced after H&E, H, or E staining. However, the structural imaging contrasts based on linear retardance did not change significantly or the staining did not generate linear birefringence on the sample area without collagen. Therefore, it can be acceptable to prepare H&E stained slices for clinical applications of polarimetry based on such a mapping relationship.

Uniaxial orientation of cellulose nanocrystals by zone-casting technique

Cellulose nanocrystals (CNCs) are biomass-based nanoparticles with attractive properties. Using a zone-casting technique, transparent films 2 cm wide and 4 cm long, and 2 µm thick, with uniaxially oriented CNCs were prepared from aqueous suspension of CNCs. The nanocrystals were aligned within the entire film perpendicular to the zone-casting direction. The orientation of the CNCs was confirmed by polarized light microscopy, X-ray diffraction and atomic force microscopy. The intensity of the light transmitted through the films depended on its polarization direction and was the strongest for the light polarized perpendicularly to the crystal orientation direction. The orientation of CNCs in the films resulting in optical anisotropy makes them promising materials for applications in optoelectronics.

Multimodal fluorescence microscope with fast adaptive polarimetry

Polarized light microscopy is a widely used technique to observe specimens that are optically anisotropic, or birefringent. It has a broad applicability in the study of minerals, soft materials such as polymers, complex fluids or liquid crystals, and organic tissues in biology and medicine. Most of these observations are qualitative in nature, as it is not obvious to quantify the spatial distribution of optical anisotropy of specimens. Moreover, existing commercial implementations for quantitative polarimetry are costly and slow in nature, precluding real time observation of dynamical processes. Here, we present a custom-made implementation of an optical microscope for quantitative polarimetry at the cost of a standard scientific polarizing microscope. The instrument allows to extract the local optical axis and birefringence of transparent materials with a frequency of several Hz. The instrument is built using off-the-shelf optomechanical components, which optimizes cost, availability, and modularity. An example of the latter is the fact that we combine the polarimetry measurements with simultaneous fluorescence microscopy, which results in a powerful multimodal instrument with broad potential applications.

Physicochemical characteristics of airborne microplastics of a typical coastal city in the Yangtze River Delta Region, China

Airborne microplastics (MPs) are important pollutants that have been present in the environment for many years and are characterized by their universality, persistence, and potential toxicity. This study investigated the effects of terrestrial and marine transport of MPs in the atmosphere of a coastal city and compared the difference between daytime and nighttime. Laser direct infrared imaging (LDIR) and polarized light microscopy were used to characterize the physical and chemical properties of MPs, including number concentration, chemical types, shape, and size. Backward trajectories were used to distinguish the air masses from marine and terrestrial transport. Twenty chemical types were detected by LDIR, with rubber (16.7%) and phenol-formaldehyde resin (PFR; 14.8%) being major components. Three main morphological types of MPs were identified, and fragments (78.1%) are the dominant type. MPs in the atmosphere were concentrated in the small particle size segment (20–50 µm). The concentration of MPs in the air mass from marine transport was 14.7 items/m3 – lower than that from terrestrial transport (32.0 items/m3). The number concentration of airborne MPs was negatively correlated with relative humidity. MPs from terrestrial transport were mainly rubber (20.2%), while those from marine transport were mainly PFR (18%). MPs in the marine transport air mass were more aged and had a lower number concentration than those in the terrestrial transport air mass. The number concentration of airborne MPs is higher during the day than at night. These findings could contribute to the development of targeted control measures and methods to reduce MP pollution.

A Study on the Materials Used in the Ancient Architectural Paintings from the Qing Dynasty Tibetan Buddhist Monastery of Puren, China.

Situated in the village of Lama Temple on the eastern bank of the Wulie River in Chengde, Puren Temple stands as one of the few remaining royal temples of great importance from the Kangxi era (1662-1722 AD). This ancient edifice has greatly contributed to the advancement of our comprehension regarding the art of royal temple painting. The present study undertakes a comprehensive analysis and identification of nine samples obtained from the beams and ceiling paintings within the main hall of Puren Temple. Furthermore, a systematic examination of their mineral pigments and adhesives is conducted. The findings from polarized light microscopy (PLM), energy-type X-ray fluorescence spectrometer (ED-XRF), micro-Raman spectroscopy (m-RS), and X-ray diffractometer (XRD) analyses reveal that the pigments present in the main hall beams of Puren Temple are cinnabar, lead white, lapis lazuli, and lime green, while the pigments in the ceiling paintings consist of cinnabar, staghorn, lead white, lapis lazuli, and lime green. The use of animal glue as a binder for these pigments on both the main hall beams and ceiling paintings is confirmed via pyrolysis-gas chromatography-mass spectrometry (Py-Gc/Ms) results. These findings hold significant implications for the future restoration of Puren Temple, as they provide valuable guidance for the selection of appropriate restoration materials.

Automatic classification of electrospun fiber polarized light images using mueller matrix depolarization parameter

Electrospun fibers are widely used in various fields of biology, medicine, and chemistry due to their unique morphological characteristics that determine their distinct application properties. Accurate and rapid classification of these fibers based on their morphology is critical for their effective utilization. Non-destructive and low-cost imaging methods are highly desirable for this purpose, so we obtained the polarization images of different forms of electrospun fibers (smooth surfaces, microporous, and beaded microspheres) by polarized light microscopy. In this study, we have explored the automatic classification of electrospun fibers based on their Mueller matrix depolarization parameter, which is highly correlated with the rough microporous structures on the surface of the object. To achieve this, we employed transfer learning and various convolutional neural networks (CNNs). Our proposed method outperformed the conventional approach that only utilizes a single Mueller matrix M44 image for classification, thus enabling researchers to effectively classify electrospun fibers. Given the high accuracy of our method, it may find significant utility in fields such as material science, nanotechnology, and bioengineering.

Characterization of the Stone Masonries and Evaluation of the Environmental Impact in Panamá Viejo: A Contribution for the Conservation of the Monumental Complex

A fundamental step in the best preservation of heritage materials is investigating their chemical and physical characteristics, and understanding how they interact with the surrounding environment. For this reason, the stone masonry of the monumental UNESCO site of Panamá Viejo was subjected to a thorough study to characterize the materials and evaluate their state of conservation. Additionally, potential outcrops were explored in order to identify sites for the supply of raw materials. The methodological approach encompassed mineralogical-petrographic investigations using stereomicroscopy, polarized light microscopy (PLM), X-ray powder diffraction (XRPD), scanning electron microscopy and energy dispersive X-ray spectrometry (ESEM-EDS) and X-ray fluorescence (XRF). The results revealed that the masonries primarily consists of polygenic breccia, basaltic andesite, rhyodacite, tuffite, and rhyolite. In addition, at the potential quarries, breccia and basaltic andesite were identified. The porous structure was studied using mercury intrusion porosimetry (MIP). Possible issues due to salt dissolution/crystallization cycles were considered. Consequently, ion chromatography (IC) was conducted on samples exhibiting alteration patinas to study the presence of soluble salts. In assessing the state of conservation, prevalent forms of deterioration included biological colonization, detachments, material loss, potential salt weathering, and chromatic alteration. In conclusion, the outcomes of this work provide a valuable resource for the current and future preservation of this site.