Laser Scanning Microscopy Research Articles

Comparative study about the results of HAZ physical simulations on different high-strength steel grades

With continuous improvements, structural steels are available in even higher strength grades above 1000 MPa yield strength. As the great majority of these steels are used in welded structures, their weldability needs to be taken into account. Several factors can cause difficulties during welding of these steels, but in this paper the softening behavior and the toughness characteristics of the heat-affected zone (HAZ) are examined. As the critical parts of the HAZ in a real welded joint are relatively small, their investigating ability is limited. However, the physical simulation provides a way of evaluating specimens made from a given material to produce the specified HAZ areas in a suitable size range for subsequent testing. In this research work, three different strength categories of high-strength structural steels (with yield strength of 960 MPa, 1100 MPa, and 1300 MPa) are investigated by physical simulation. In the case of different technological variants of gas metal arc welding (GMAW) process, the effect of the cooling time t8/5 is investigated in different HAZ subzones considered to be critical. The thermal cycles were determined according to the Rykalin 3D model. The investigated cooling times were t8/5 = 5 s, 15 s, and 30 s. The properties of the selected coarse-grained, intercritical and intercritically reheated coarse-grained zones are analyzed by laser scanning microscope, scanning electron microscope, hardness test, and instrumented Charpy V-notch impact toughness test. Furthermore, additional investigation like JMatPro calculations, electron backscatter diffraction measurements, and prior austenite grain size calculation were carried out. As a result of the tests, the investigated heat-affected subzones indicated higher sensitivity to the welding heat input compared to conventional structural steels. Overall, the results of the tests show that the application of shorter t8/5 cooling time can be beneficial for the investigated high-strength steel grades, since significant toughness reduction and the risk of softening occur in the whole cooling time range.

Biocompatibility of Bioactive Sealers Bio-C Sealer vs MTA Repair HP in Human Fibroblasts

Bioactive cements based on tricalcium silicate have been introducedto the market for use in dentistry, with a variety of clinical applications. These cements are in contact with vital tissues such as dental pulp or periodontium in cases of unintentional extrusion; thus, it is important to know the genotoxicity and cytoxicity of these materials. The objective of this study was to evaluate the cytotoxicity and genotoxicity of bioactive sealers, Bio-C® Sealer and MTA Repair HP®, in human fibroblasts. Discs of bioactive sealers Bio-C® Sealer, and MTA Repair HP®, were prepared and set for 24h under sterile conditions. The discs were placed in culture medium at 2.5mg/mL inside a SRT6D roller mixer (Stuart, UK) at 60rpm for 24h. The eluates obtained were incubated for 24h with previously activated and cultured ATCC cell line fibroblasts at 80% confluence. The cytotoxicity was evaluated by Alamar Blue® and LIVE/DEAD assays, as well as the analysis of the Tunel and Mitotracker assays to evaluate genotoxicity using the confocal laser-scanning microscope. In the Alamar Blue® assay, the Bio-C® Sealer presented a cell proliferation of 87%, while the MTA Repair HP® Sealer was 72%. A statistically significant difference was found between the MTA Repair HP® Sealant and the negative control (p=<0.001). Regarding the genotoxicity tests, in the Tunel assay, both materials stain the nucleus of the fibroblast cells exposed to the eluates, while in the Mitotracker assay, the MTA Repair HP® Sealer showed greater mitochondrial function than the Bio-C® Sealer. Calcium silicate-based sealers, Bio-C® Sealer and MTA Repair HP®, are not cytotoxic and have low genotoxicity.

Advanced spectroscopic investigation of colour centres in LiF crystals irradiated with monochromatic hard x-rays

Nominally-pure lithium fluoride (LiF) crystals were irradiated with monochromatic hard x-rays of energy 5, 7, 9 and 12 keV at the METROLOGIE beamline of the SOLEIL synchrotron facility, in order to understand the role of the selected x-ray energy on their visible photoluminescence (PL) response, which is used for high spatial resolution 2D x-ray imaging detectors characterized by a wide dynamic range. At the energies of 7 and 12 keV the irradiations were performed at five different doses corresponding to five uniformly irradiated areas, while at 5 and 9 keV only two irradiations at two different doses were carried out. The doses were planned in a range between 4 and 1.4 × 103 Gy (10.5 mJ cm−3 to 3.7 J cm−3), depending on the x-ray energy. After irradiation at the energies of 7 and 12 keV, the spectrally-integrated visible PL intensity of the F2 and F3 + colour centres (CCs) generated in the LiF crystals, carefully measured by fluorescence microscopy under blue excitation, exhibits a linear dependence on the irradiation dose in the investigated dose range. This linear behaviour was confirmed by the optical absorption spectra of the irradiated spots, which shows a similar linear behaviour for both the F2 and F3 + CCs, as derived from their overlapping absorption band at around 450 nm. At the highest x-ray energy, the average concentrations of the radiation-induced F, F2 and F3 + CCs were also estimated. The volume distributions of F2 defects in the crystals irradiated with 5 and 9 keV x-rays were reconstructed in 3D by measuring their PL signal using a confocal laser scanning microscope operating in fluorescence mode. On-going investigations are focusing on the results obtained through this z-scanning technique to explore the potential impact of absorption effects at the excitation laser wavelength.

Sucrose-phosphate osmotic system improves the quality characteristics of reduced-salt salted egg yolk: Profiling from protein structure and lipid distribution perspective

This paper was dedicated to the study of the effect of sucrose-phosphate on aspects of physicochemical properties, lipid distribution and protein structure during the picklig of reduced‐salt salted egg yolk (SEY). This work constructed a reduced‐salt pickling system from a new perspective (promoting osmosis) by using a sucrose-phosphate-salt. Results showed that SEY-28d achieved a desirable salt content (1.07 %), hardness (573.46 g) and springiness (0.65 g). The matured SEY was in excellent quality with orange-red color and loose sandy texture. This was because the lipoprotein aggregated with each other through hydrophobic interaction to form a stable network structure. In addition, the hypertonic environment accelerated salt penetration. These also created good condition for lipid spillage. The results of confocal laser scanning microscope also verified this phenomenon. This work provides important guidance for new reduced‐salt curing of traditional pickled foods, deep processing of SEY, and industry development in the field of poultry egg.

Experiment of droplet contact angle hysteresis on PTFE fibrous membrane in pore-scale

The present paper aims to study the influence of pore-scale fiber structure on the Cassie-Baxter contact angle of droplets on PTFE fibrous hydrophobic membranes. The fiber structure is obtained by the Confocal Laser Scanning Microscope (CLSM), and an optical test bench is built to measure the Cassie-Baxter contact angles of a droplet on a PTFE membrane. The free energy of the solid-liquid interface is measured by the surface tension integral on the droplet profile. An iterative algorithm for liquid-vapor interface shrinking is brought forward to analyze the liquid-vapor interface area between the droplet and the membrane, and the interface parameters of the Cassie-Baxter model are recognized. The experiment shows that the Cassie-Baxter contact angles and the contact line radii along the droplet circumference are in a normal distribution due to random fiber structures, and the variance of the normal distribution increases with the membrane pore diameter. The normal distribution of Cassie-Baxter’s contact angle indicates the constraint of the solid-liquid interface to the liquid-vapor interface. On the same normal contact pressure condition, the liquid-vapor area ratio of the droplet and membrane interface changes little, and Cassie-Baxter’s contact angle increases with the solid-liquid interface area ratio.

Antimicrobial and anti-biofilm activity of a thiazolidinone derivative against Staphylococcus aureus in vitro and in vivo.

Staphylococcus aureus (S. aureus) causes many infections with significant morbidity and mortality. S. aureus can form biofilms, which can cause biofilm-associated diseases and increase resistance to many conventional antibiotics, resulting in chronic infection. It is critical to develop novel antibiotics against staphylococcal infections, particularly those that can kill cells embedded in biofilms. This study aimed to investigate the bacteriocidal and anti-biofilm activities of thiazolidinone derivative (TD-H2-A) against S. aureus. A total of 40 non-duplicate strains were collected, and the minimum inhibitory concentrations (MICs) of TD-H2-A were determined. The effect of TD-H2-A on established S. aureus mature biofilms was examined using a confocal laser scanning microscope (CLSM). The antibacterial effects of the compound on planktonic bacteria and bacteria in mature biofilms were investigated. Other characteristics, such as cytotoxicity and hemolytic activity, were researched. A mouse skin infection model was used, and a routine hematoxylin and eosin (H&E) staining was used for histological examination. The MIC values of TD-H2-A against the different S. aureus strains were 6.3-25.0 µg/mL. The 5 × MIC TD-H2-A killed almost all planktonic S. aureus USA300. The derivative was found to have strong bacteriocidal activity against cells in mature biofilms meanwhile having low cytotoxicity and hemolytic activity against Vero cells and human erythrocytes. TD-H2-A had a good bacteriocidal effect on S. aureus SA113-infected mice. In conclusion, TD-H2-A demonstrated good bacteriocidal and anti-biofilm activities against S. aureus, paving the way for the development of novel agents to combat biofilm infections and multidrug-resistant staphylococcal infections.IMPORTANCEStaphylococcus aureus, a notorious pathogen, can form a stubborn biofilm and develop drug resistance. It is crucial to develop new anti-infective therapies against biofilm-associated infections. The manuscript describes the new antibiotic to effectively combat multidrug-resistant and biofilm-associated diseases.

Improving the surface quality of AlMgSi1 alloy with the selection of the appropriate vibration grinding stones

The number and variety of paints and coating systems in the automotive industry have increased in recent years. However, the basic requirement for the quality of the paints still depends on the surfaces on which they are applied. This applies not only to the large body parts but also to all components that are integrated.There are many surface treatment processes and techniques that are used as an additional process step, such as pickling for a chemical surface treatment or blasting for a mechanical surface treatment.The present work investigates a method for optimizing the surface before painting, using the membrane cup of the ultrasonic sensor as an example for AlMgSi1 alloy, without implementing additional process steps.First, all process steps influencing the surface quality for the production of a membrane cup were considered for optimization. Then, based on a backward process chain analysis, vibratory grinding for deburring of the membrane cup was determined as the process step to be optimized. For the optimization of the vibratory grinding, experiments with two different grindstones were performed. For the characterization of the surface quality, the roughness parameters Rz and Ra were measured with a confocal laser scanning microscope and analyzed in order to determine the optimal process conditions.The experiments showed that the surface roughness can be significantly improved using dedicated process parameters and taking into account the geometry of the vibratory grinding stones. Moreover, different surface qualities can be achieved at different areas on the workpiece, this new approach provides a solution for surface optimization in various areas of the workpiece without additional process steps and costs.

Evaluating the Effect of Cement and ARG Fiber on the Mechanical and Microstructural Properties of Dune Sand

Despite its collapsible nature and weakness, desert sand can be used for construction purposes all over the world if properly stabilized. Therefore, the aim of this study is to evaluate the effectiveness of cement and fiber in stabilizing locally available dune sand. A test plan was used to investigate the effects of varying quantities of alkali resistance glass (ARG) fiber (F: 0%, 0.2%, 0.4%, and 0.6%) and portland cement (C: 0.0%, 1.0%, 3.0%, and 5.0%) on the mechanical and microstructural properties of dune sand. Mechanical properties such as unconfined strength (UCS), strain at failure (εf), California bearing ratio (CBR), and modulus of elasticity (Es) were evaluated, and microstructure properties were investigated using Raman spectrum and laser-scanning microscopy (LSM) tests on stabilized sand samples. The results of the experimental study showed that the percentage of cement in the treated sand has a more significant impact on the investigated properties of the treated sand than the percentage of fibers. In addition, increasing fiber content results in an increase in the ductility of the sand mix. Raman analysis revealed significant interactions between sand mix components. Moreover, LSM results showed that fiber–cement interaction increased with increasing cement percentage, as calcium silicate hydrates (CSH) formed in the mix and filamentous and intrastrand binding occurred. The findings of this study indicate that ARG fiber and cement can be effective in the stabilization of dune sand for construction purposes even with the use of low percentages of ARG fiber (0.2%–0.4%) and cement (3%).

Is a mix - A fix? "A microscopic analysis of depth of penetration of three combinations of irrigants".

The aim of the study is to evaluate the penetration depth of irrigants mixed with NaOCl into dentinal tubules at the apical third of the root canal using a confocal laser scanning microscope (CLSM). Thirty-six single-rooted teeth were selected, decoronated, and straight-line access established. Then, the canal was enlarged with hand Protaper till size F3. The samples were divided randomly into three groups: Group 1 was irrigated with 3% NaOCl mixed with 1% phytic acid; Group 2 was irrigated with 3% NaOCl mixed with 18% etidronic acid; and Group 3 was irrigated with 3% NaOCl mixed with chitosan. The samples were subjected to CLSM evaluation. One-way analysis of variance with Tukey's post hoc was used for statistical analysis. Group 3 (NaOCl with chitosan) showed a greater depth of penetration in comparison to Groups 1 and 2. Within limitations, it can be concluded that chitosan, in combination with NaOCl can be used as an alternative to the current irrigation protocol.

Phase-field simulation and high-temperature confocal laser scanning microscope experimental study of Martensite transformation during the disk laser quenching process

Laser quenching has significant advantages over conventional heat treatment. The main microstructure of C30E4 steel after quenching at a specific temperature is Martensite. Accurately obtaining the microscopic characteristics of the Martensite transformation process is the key to obtaining the best material properties, which is significant to material design. In this paper, the C30E4 steel was disk-laser-quenched, the metallographic and high-temperature confocal laser scanning microscope experiment was carried out. The microstructure of the fully phase-change hardened zone, heat-affected transition zone and matrix zone after quenching was obtained. The C30E4 martensitic transformation process in-situ was obtained. The mathematical physics model of Martensite transformation from metastable to steady state was established by phase field method. The finite-element method was used to solve the model, and the quenching Martensite transformation law was obtained. The Martensite formation process in austenite grain boundaries and microstructure defects was discussed. The experimental and the numerical simulation were compared to get consistent results, which verify the validity of the calculation method. This study lays a significant foundation for effectively revealing the Martensite formation and transformation during the disk laser quenching process, and provides a theoretical basis for quantifying the influence of Martensite transformation process on material properties.

What Is the Impact of Antimicrobial Photodynamic Therapy on Oral Candidiasis? An In Vitro Study.

This study aimed to evaluate the ability of photodynamic therapy, based on the use of a gel containing 5% delta aminolaevulinic acid (ALAD) for 45' followed by irradiation with 630 nm LED (PDT) for 7', to eradicate Candida albicans strains without damaging the gingiva. C. albicans oral strains and gingival fibroblasts (hGFs) were used to achieve these goals. The potential antifungal effects on a clinical resistant C. albicans S5 strain were evaluated in terms of biofilm biomass, colony forming units (CFU/mL) count, cell viability by live/dead analysis, and fluidity membrane changes. Concerning the hGFs, viability assays, morphological analysis (optical, scanning electronic (SEM), and confocal laser scanning (CLSM) microscopes), and assays for reactive oxygen species (ROS) and collagen production were performed. ALAD-mediated aPDT (ALAD-aPDT) treatment showed significant anti-biofilm activity against C. albicans S5, as confirmed by a reduction in both the biofilm biomass and CFUs/mL. The cell viability was strongly affected by the treatment, while on the contrary, the fluidity of the membrane remained unchanged. The results for the hGFs showed an absence of cytotoxicity and no morphological differences in cells subjected to ALAD-aPDT expected for CLSM results that exhibited an increase in the thickening of actin filaments. ROS production was augmented only at 0 h and 3 h, while the collagen appeared enhanced 7 days after the treatment.

Polyelectrolyte Multilayer Coating on 3D Printed PEGDA/TCP Scaffold with Improved Cell Proliferation

Bioactive coating of ceramic scaffolds is an effective way to ameliorate osseointegration and attenuate implant-induced inflammatory responses, which should be biocompatible and possess suitable mechanical properties to regulate cell adhesion and migration. In this study, a poly (ethylene glycol) diacrylate/tricalcium phosphate (PEGDA/TCP) ceramic scaffold was prepared using SLA-3D printing, and its compressive strength was 8.9 ± 1.0 MPa. Chitosan (Chi) and chondroitin sulfate (CS) were assembled on the surface of the PEGDA/TCP scaffolds and crosslinked with 1-ethyl-3-(3-dimethylaminopropyl) -carbodiimide/ N-hydroxysuccinimide (EDC/NHS). Scanning electron microscope (SEM), Fourier transform infrared (FTIR), and laser scanning microscope were used to evaluate the surface modification of the PEGDA/TCP scaffolds. Cellular tests showed that polyelectrolyte multilayers (PEMs) promoted cell adhesion and proliferation of osteoblasts relative to unmodified scaffolds. Furthermore, it can be demonstrated that the SLA-3D printed TCP scaffolds could meet the compressive requirements of trabecular bones, and the bioactivity of the bone scaffolds could be effectively improved by combining them with Chi/CS PEM.

Discovery of Melittin as Triple-Action Agent: Broad-Spectrum Antibacterial, Anti-Biofilm, and Potential Anti-Quorum Sensing Activities.

The development of antibiotic-resistant microorganisms is a major global health concern. Recently, there has been an increasing interest in antimicrobial peptides as a therapeutic option. This study aimed to evaluate the triple-action (broad-spectrum antibacterial, anti-biofilm, and anti-quorum sensing activities) of melittin, a membrane-active peptide present in bee venom. The minimum inhibitory concentration and minimum bactericidal concentration of the melittin were determined using the microdilution method and agar plate counting. Growth curve analysis revealed that melittin showed a concentration-dependent antibacterial activity. Scanning electron microscope analysis revealed that melittin treatment altered the morphology. Confocal laser scanning microscope revealed that melittin increased the membrane permeability and intracellular ROS generation in bacteria, all of which contribute to bacterial cell death. In addition, the crystal violet (CV) assay was used to test the anti-biofilm activity. The CV assay demonstrated that melittin inhibited biofilm formation and eradicated mature biofilms. Biofilm formation mediated by quorum sensing (QS) plays a major role in this regard, so molecular docking and molecular dynamics analysis confirmed that melittin interacts with LasR receptors through hydrogen bonds, and further evaluates the anti-QS activity of melittin through the production of virulence factors (pyocyanin, elastase, and rhamnolipid), exopolysaccharides secretion, and bacterial motility, that may be the key to inhibiting the biofilm formation mechanism. The present findings highlight the promising role of melittin as a broad-spectrum antibacterial, anti-biofilm agent, and potential QS inhibitor, providing a new perspective and theoretical basis for the development of alternative antibiotics.

Inactivation activity and mechanism of pulsed light against Alicyclobacillus acidoterrestris vegetative cells and spores in concentrated apple juice

Alicyclobacillus acidoterrestris has received much attention due to its unique thermo-acidophilic property and implication in the spoilage of pasteurized juices. The objective of this study was to evaluate the sterilization characteristics and mechanisms of pulsed light (PL) against A. acidoterrestris vegetative cells and spores in apple juice. The results indicated that bacteria cells in apple juice (8–20°Brix) can be completely inactivated within the fluence range of 20.25–47.25 J/cm2, which mainly depended on the soluble solids content (SSC) of juice, and the spores in apple juice (12°Brix) can be completely inactivated by PL with the fluence of 54.00 J/cm2. The PL treatment can significantly increase the leakage of reactive oxygen species (ROS) and proteins from cells and spores. Fluorescence studies of bacterial adenosine triphosphate (ATP) indicated that the loss of ATP was evident. Scanning electron microscopy and confocal laser scanning microscope presented that PL-treated cells or spores had serious morphological damage, which reduced the integrity of cell membrane and led to intracellular electrolyte leakage. In addition, there were no significant negative effects on total sugars, total acids, total phenols, pH value, SSC and soluble sugars, and organic acid content decreased slightly during the PL treatment. The contents of esters and acids in aroma components had a certain loss, while that of alcohols, aldehydes and ketones were increased. These results demonstrated that PL treatment can effectively inactivate the bacteria cells and spores in apple juice with little effect on its quality. This study provides an efficient method for the inactivation of A. acidoterrestris in fruit juice.

Characterizing Nanoparticle Isolated by Yam Bean (Pachyrhizus erosus) as a Potential Agent for Nanocosmetics: An in vitro and in vivo Approaches.

This study investigated the potential of Plant-Derived Exosome-Like Nanoparticles (PDENs) as cosmeceutical nanocarriers for treating skin problems, such as scar removal, face rejuvenation, anti-aging, and anti-pigmentation. Researchers isolated PDENs from Yam Bean (Pachyrhizus erosus) using PEG-based precipitation, gradual filtration, and various centrifugations at low temperatures. Followed by in vitro and in vivo studies using HDF cells and Zebrafish. The morphology of the YB-PDENs was determined using TEM analysis, they had a spherical shape with diameters of 236,83 ± 9,27nm according to PSA. The study found that YB-PDENs were stable in aquabidest at 4°C for one month of storage and had ~-26,5 mV of Zeta Potential. The concentration of YB-PDENs was measured using the BCA Assay, and internalization of YB-PDENs to HDF cells was observed using a Confocal Laser Scanning Microscope labelled with PKH67. As for cytotoxicity, after 24 and 72 hours of incubation with YB-PDENs, the viability of HDF cells remained more than 80%. The study also examined cell migration using the Scratch Assay and found that at 2,5 μg/mL, YB-PDENs had better migration results than other concentrations. Immunocytochemistry showed that collagen expression was higher after 14 days of incubation with YB-PDENs, and melanocytes in zebrafish decreased at each concentration compared with controls. In conclusion, this study is the first to extract and describe PDENs from Yam Bean (Pachyrhizus erosus), with YB-PDENs having a promising anti-melanogenic effect in skin treatment. This study highlights the potential of YB-PDENs as a promising alternative to depigmentation and skin whitening treatments.

Swept-wavelength null polarimetry for highly sensitive birefringence laser scanning microscopy

We have recently demonstrated a high-speed null polarimeter [Opt. Express 30, 18889 (2022)10.1364/OE.454193OPEXFF1094-4087] based on passive polarization optics and using a fast swept-wavelength laser source. We report here its implementation in a laser-scanning microscope setup, enabling highly sensitive linear retardance imaging with a pixel dwell time of 10 μs. The instrument is also able to measure light depolarization induced by the sample. Images of biological samples, including cancerous tissue and cells, illustrate its performances.

Comparison of 2D and 3D measurement methods for evaluating laser structured aluminum surfaces using fractal dimension

The interlaminar strength of mechanically interlocked polymer-metal hybrids is strongly determined by the surface geometry at the interface. To improve the interlaminar strength, laser beam machining is among the most applied technologies for manufacturing a predefined and complex surface structure. Previously, the fractal dimension of interfaces in polymer-metal-hybrids has been introduced as scale-independent measure. It has been correlated quantitatively with the interlaminar strength using values derived from cross-sectional images. Therefore, the quality of the prediction directly depends on the accuracy of the measured surface data. However, a holistic acquisition of surface structure for the determination of the fractal dimension including 3D methods has not been investigated so far. In this study, laser machining was used to generate five different pin-structures on samples of the aluminum alloy EN AW-6082. Surface characterization was realized by light microscopy (2D), laser scanning microscopy (3D) and X-ray microscopy (3D). Subsequently, the fractal dimension was determined using a box-counting algorithm as well as slit-island method. The results show that all methods regarded are appropriate to determine the fractal dimension, but differ in the specific results. The functional characteristics of each method are described in detail and potentials for applications are outlined.

Influence of the width of cut in ultrasonic vibration superimposed face milling of X46Cr13 and X40CrMoV5-1 on the surface microstructure and CVD diamond coating adhesion

Ultrasonic vibration superimposed milling (UVSM) with an oscillating movement of the specimen in the direction of the tool axis is an efficient method for the generation of microstructured functional surfaces. The application of end milling cutters with a diameter smaller than the length and the width of the surface to be structured generally results in recuts and path overlaps. In order to avoid surface recutting, a small lead angle was realized by slightly tilting the ultrasonic system with the specimen. The overlapping area of two adjacent paths was kinematically simulated for different widths of cut (from 70% to 90% of the tool diameter). Cutting experiments were carried out by UVSM of X46Cr13 and X40CrMoV5-1 steel specimens to investigate the influence of the width of cut on the surface generated in the overlapping area and also the adherence of a subsequently deposited CVD diamond layer. For this purpose, 3-D laser scanning microscopy and SEM analysis were applied. The knowledge gained allows for designing UVSM processes, especially regarding 3-D freeform surfaces with defined microstructures subjected to CVD diamond coating.

Confocal Microscopy of Fluorescently Labeled EHDV-Infected Cell Cultures.

The confocal laser scanning microscope allows the visualization of intracellular structures in greater detail than a widefield fluorescence microscope. Immunofluorescence (IF) techniques make use of the inherent ability of antibodies to bind to specific epitopes of specific proteins. Tagging these antibodies with an easily visualized molecule, e.g., a fluorophore, enables imaging in the fluorescence microscope. This is, however, a localization technique and will only give information about where certain proteins are; it does not provide the ultrastructural context provided by the transmission electron microscope. It also relies heavily on the accuracy and binding affinity of individual primary antibodies. Despite this, it is a commonly used, robust, and adaptable technique. In this chapter, we use a long-established IF protocol from our laboratory to locate EHDV proteins in a monolayer of infected cultured cells.

Er,Cr:YSGG and 980nm diode lasers influence dentin surface volume after cariogenic challenge: in vitro study.

This study aimed to evaluate the influence of the Er,Cr:YSGG irradiation and 980-nm diode lasers on the surface roughness (SR) and volume loss (VL) of dentin subjected to cariogenic challenge. Subsequently, 130 specimens of bovine dentin were divided into the following 13 groups: NT: no treatment; FG: fluoride gel; FV: fluoride varnish; Di: 980-nm diode; Di + FG; Di + FV; FG + D; FV + Di; Er: Er,Cr:YSGG; Er + FG; Er + FV; FG + Er and FV + Er. Er,Cr:YSGG laser parameters were as follows: 0.25 W; 5.0 Hz; 4.46 J/cm2 without water and 55% air. Furthermore, the 980-nm diode laser parameters were 2.0 W; 2.0 Hz; 21.41 J/cm2. The samples from each group were subjected to pH cycling. A confocal laser scanning microscope was used to evaluate SR and VL. Difference between the volume of the reference and treated areas + DES/RE was used to determine SR and VL. The mean values of the different groups were subjected to analysis of variance and Tukey's post-hoc test. The VL values were analyzed using the Kruskal-Wallis and Dunn post-hoc test (p < 0.05). The SR of the reference area did not show a statistically significant 1807-3107-bor-38-e025treatment and cariogenic challenge (p > 0.05). Moreover, VL in the FV + Di and FV + Er groups showed a statistically significant difference compared with areas submitted to different types of treatment and cariogenic challenge (p > 0.05). Er,Cr:YSGG and 980-nm diode lasers associated with fluoride varnishes decreased dentin VL in bovine teeth submitted to cariogenic challenge.