Laser Scanning Microscopy Research Articles

Red and Green Laser Powder Bed Fusion of Pure Copper in Combination with Chemical Post-Processing for RF Cavity Fabrication

Linear particle accelerators (Linacs) are primarily composed of radio frequency cavities (cavities). Compared to traditional manufacturing, Laser Powder Bed Fusion (L-PBF) holds the potential to fabricate cavities in a single piece, enhancing Linac performance and significantly reducing investment costs. However, the question of whether red or green laser PBF yields superior results for pure copper remains a subject of ongoing debate. Eight 4.2 GHz single-cell cavities (SCs) were manufactured from pure copper using both red and green PBF (SCs R and SCs G). Subsequently, the surface roughness of the SCs was reduced through a chemical post-processing method (Hirtisation) and annealed at 460 °C to maximize their quality factor (Q0). The geometric accuracy of the printed SCs was evaluated using optical methods and resonant frequency (fR) measurements. Surface conductivity was determined by measuring the quality factor (Q0) of the SCs. Laser scanning microscopy was utilized for surface roughness characterization. The impact of annealing was quantified using Energy-Dispersive X-ray Spectroscopy and Electron Backscatter Diffraction to evaluate chemical surface properties and grain size. Both the SCs R and SCs G achieved the necessary geometric accuracy and thus fR precision. The SCs R achieved a 95% Q0 after a material removal of 40 µm. The SCs G achieved an approximately 80% Q0 after maximum material removal of 160 µm. Annealing increased the Q0 by an average of about 5%. The additive manufacturing process is at least equivalent to conventional manufacturing for producing cavities in the low-gradient range. The presented cavities justify the first high-gradient tests.

Enhanced Stability and Detection Range of Microbial Electrochemical Biotoxicity Sensor by Polydopamine Encapsulation.

With the rapid development of modern industry, it is urgently needed to measure the biotoxicity of complex chemicals. Microbial electrochemical biotoxicity sensors are an attractive technology; however, their application is usually limited by their stability and reusability after measurements. Here, we improve their performance by encapsulating the electroactive biofilm with polydopamine (PDA), and we evaluate the improvement by different concentrations of heavy metal ions (Cu2+, Ag+, and Fe3+) in terms of inhibition ratio (IR) and durability. Results indicate that the PDA-encapsulated sensor exhibits a more significant detection concentration than the control group, with a 3-fold increase for Cu2+ and a 1.5-fold increase for Ag+. Moreover, it achieves 15 more continuous toxicity tests than the control group, maintaining high electrochemical activity even after continuous toxicity impacts. Images from a confocal laser scanning microscope reveal that the PDA encapsulation protects the activity of the electroactive biofilm. The study, thus, demonstrates that PDA encapsulation is efficacious in improving the performance of microbial electrochemical biotoxicity sensors, which can extend its application to more complex media.

Deep learning-based prediction of 3-dimensional silver contact shapes enabling improved quality control in solar cell metallization

The industrial metallization of Si solar cells predominantly relies on screen printing, with silver as the preferred electrode material. However, the design of commercial screens often leads to suboptimal silver usage and increased electrical resistance due to print-related inhomogeneities like mesh marks, constrictions and spreading. Real-time monitoring of quality parameters during production has thus become increasingly critical. Current inline optical quality control systems usually only include 2D visualizations of the printed layout, which limits their effectiveness in quality control. Options that allow 3D measurements are usually slow, expensive, and therefore not worth considering in most cases. This research focuses on the development of a model that can estimate the three-dimensional shape of printed contact fingers from a single 2D image without the need of additional hardware using deep learning. Furthermore, a workflow for the generation of training data, which involves the creation of image pairs from a 2D microscope and a 3D confocal laser scanning microscope (CLSM) to accurately represent solar cell fingers, is presented. After model training, the predicted height maps are compared with the ground truth height maps, and the robustness of the model with respect to a paste variation and screen parameter variation is examined. The results confirm the feasibility and reliability of deep learning-based 3D shape estimation, extending its applicability to new, previously unseen data from screen-printed contact fingers. With a structural similarity index (SSIM) score of 0.76, a strong correlation between the estimated and ground truth height maps is established. In summary, our deep learning-based approach for height map estimation offers an effective and reliable solution for fast inline detection and analysis of the cross-sectional area of the printed contact fingers.

Utilization of mycelial polysaccharide from Schizophyllum commune for the formation of highly stable o/w emulsion

This study investigated the emulsifying properties of a hot water extracted polysaccharide (SCMP) derived from liquid fermentation mycelium of Schizophyllum commune in stabilizing oil-in-water (o/w) emulsions. SCMP demonstrated the ability to stabilize o/w emulsions (referred to as SCMPe) for extended periods (>21 days) even at low concentrations (2 g/L) and increased the viscosity and elasticity of the emulsion system. Through confocal laser-scanning microscope visualization, it was observed that SCMP created a physical barrier on the surface of the oil droplets, contributing to emulsion stability. Furthermore, SCMPe demonstrated excellent tolerance to varying concentrations of Na+ (0.1–0.5 mol/L) and Ca2+ (0.01–0.05 mol/L) as well as a wide pH range (3.0–9.0). Moreover, increasing SCMP concentration effectively inhibited lipid oxidation and improved the stability of β-carotene and curcumin in the emulsion during storage. A detailed analysis of SCMP's composition and structure was done to better understand its emulsifier properties, revealing SCMP to be composed of two β-glucans with a rigid chain and high molecular weight. These results suggest that SCMP holds great potential an efficient stabilizer in the food industry, particularly in enhancing emulsion stability.

4D Single-particle tracking with asynchronous read-out single-photon avalanche diode array detector

Single-particle tracking techniques enable investigation of the complex functions and interactions of individual particles in biological environments. Many such techniques exist, each demonstrating trade-offs between spatiotemporal resolution, spatial and temporal range, technical complexity, and information content. To mitigate these trade-offs, we enhanced a confocal laser scanning microscope with an asynchronous read-out single-photon avalanche diode array detector. This detector provides an image of the particle’s emission, precisely reflecting its position within the excitation volume. This localization is utilized in a real-time feedback system to drive the microscope scanning mechanism and ensure the particle remains centered inside the excitation volume. As each pixel is an independent single-photon detector, single-particle tracking is combined with fluorescence lifetime measurement. Our system achieves 40 nm lateral and 60 nm axial localization precision with 100 photons and sub-millisecond temporal sampling for real-time tracking. Offline tracking can refine this precision to the microsecond scale. We validated the system’s spatiotemporal resolution by tracking fluorescent beads with diffusion coefficients up to 10 μm2/s. Additionally, we investigated the movement of lysosomes in living SK-N-BE cells and measured the fluorescence lifetime of the marker expressed on a membrane protein. We expect that this implementation will open other correlative imaging and tracking studies.

Ultra-low-cost and high-fidelity NIR-II confocal laser scanning microscope with Bessel beam excitation and SiPM detection.

Confocal laser scanning microscopy (CLSM) is one of the most important imaging tools in the biomedical field, and near-infrared-II (NIR-II, 900-1700nm) fluorescence imaging technology has also made fruitful research progress in deep imaging in recent years. The NIR-II based CLSM has problems such as an expensive detector and reduced image resolution caused by long wavelength excitation. Here, by simultaneously using a low-cost silicon photomultiplier (SiPM) as a detector and a Bessel beam as an excitation, we developed an ultra-low-cost and high-fidelity NIR-II confocal laser scanning microscope. The use of SiPM reduces the cost of the NIR-II fluorescence detection module in CLSM, while enabling the detection of ultra-broadband fluorescence signals spanning visible to NIR-II regions. The introduction of the Bessel beam compensates to some extent for the weakening of spatial resolution caused by the increase in the wavelength of light in the NIR region. Experimental results show that the use of the Bessel beam can improve the resolution by 12% when observing thin samples. With the increase of sample thickness, the imaging resolution of the Bessel beam at NIR-II wavelengths is better than that of the Gaussian beam at NIR-I wavelengths at the penetrable depth of the NIR-I light. At deeper depths, the imaging resolution and imaging depth of Bessel beam CLSM is superior to Gaussian beam CLSM at the same excitation power.

Effect of Boron on the Solidification Characteristics and Constitutive Equation of S31254 Superaustenitic Stainless Steel

Solidification structure and segregation behavior of S31254 superaustenitic stainless steel ingot containing 0 wt% B and 0.005 wt% B are investigated. The results show that serious element segregation and massive eutectics are present in the center of the ingot. By contrast, the addition of boron reduces the secondary dendrite spacing, the degree of elemental segregation, and the σ phase. Further, the effect of boron on the solidification process is studied by using the high‐temperature confocal laser scanning microscope and combined with Thermo‐Calc thermodynamic simulation results. The results show that boron can widen the solidification temperature range and greatly retard the matrix solidification. The role of boron in refining dendrite structure and inhibiting precipitation is further confirmed. The effect of boron on the constitutive equation of S31254 superaustenitic stainless steel has also been researched through isothermal compression testing in the temperature range of 950–1200 °C and strain rate range of 0.01–10 s−1. The activation energy of the boron‐free and boron‐containing S31254 superaustenitic stainless steels based on the constitutive equation are 427.77 and 495.80 kJ mol−1, respectively. Processing maps indicate that the instability domain is significantly reduced and the hot ductility is improved after the addition of B.

The combined effect of acid and mild heat treatment on the shelf-life and quality of fresh wet noodles

In this research, changes in microbial shelf-life and quality characteristics of fresh wet noodles (FWNs) were analyzed under acid and different mild heat treatment conditions. Acid combined with heat treatment at 50 °C for 30 min extended the shelf-life of FWNs by 30–40 d and effectively delayed the color deterioration of FWNs. The water absorption of FWNs was reduced by the mild heat treatment. Meanwhile, the cooking loss of the noodles and hardness significantly (p < 0.05) increased. Differential scanning calorimetry (DSC) analysis showed that the gelatinization enthalpy of the starch in FWNs decreased and the gelatinization temperature increased. The X-ray diffraction (XRD) results showed that the crystallinity of the starch decreased from 27.51% to 24.2% after mild heat treatment. In addition, it was observed by confocal laser scanning microscope that the mild heat treatment made the gluten networks more continuous. The free sulfhydryl content of noodles showed a decreasing trend, while the sodium dodecyl sulfate-extractable protein (SDS-EP) value was also significantly (p < 0.05) reduced. Overall, the combination of mild heat treatment with acid extended the shelf-life of FWNs and had less impact on their quality.

Suppressing the virulence factors of Candida auris with baicalein through multifaceted mechanisms.

Candida auris, a rapidly spreading multi-drug-resistant fungus, is causing lethal infections under certain conditions globally. Baicalin (BE), an active ingredient extracted from the dried root of Scutellaria baicalensis Georgi, exhibits antifungal activity. However, studies have shown the distinctive advantages of Traditional Chinese medicine in combating fungal infections, while the effect of BE, an active ingredient extracted from the dried roots of Scutellaria baicalensis Georgi, on C. auris, remains unknown. Therefore, this study aims to evaluate the potential of BE as an antifungal agent against the emerging multidrug-resistant C. auris. Various assays and models, including microbroth dilution, time growth curve analysis, spot assays, adhesion tests, flocculation test, cell surface hydrophobicity assay, hydrolase activity assays, XTT assay, violet crystal assay, scanning electron microscope (SEM), confocal laser scanning microscope (CLSM), flow cytometry, Live/dead fluorescent staining, reactive oxygen species (ROS), cell wall assay, aggregation assay, porcine skin model, Galleria mellonella larvae (G. mellonella larvae) infection model, and reverse transcription-quantitative polymerase chain reaction (RT-PCR) were utilized to investigate how baicalein suppresses C. auris through possible multifaceted mechanisms. The findings indicate that BE strongly inhibited C. auris growth, adhesion, and biofilm formation. It also effectively reduced drug resistance and aggregation by disrupting the cell membrane and cell wall while reducing colonization and invasion of the host. Transcriptome analysis showed significant modulation in gene expression related to different virulence factors post-BE treatment. In conclusion, BE exhibits significant effectiveness against C. auris, suggesting its potential as a viable treatment option due to its multifaceted suppression mechanisms.

Dual-Responsive "Egg-Box" Shaped Microgel Beads Based on W1/O/W2 Double Emulsions for Colon-Targeted Delivery of Synbiotics.

In this study, for enhancing the resistance of probiotics to environmental factors, we designed a microgel beads delivery system loaded with synbiotics. Multiple droplets of W1/O/W2 emulsions stabilized with zein-apple pectin hybrid nanoparticles (ZAHPs) acted as the inner "egg," whereas a three-dimensional network of poly-L-lysine (PLL)-alginate-CaCl2 (Ca) crosslinked gel layers served as the outermost "box." ZAHPs with a mass ratio of 2:1 zein-to-apple pectin showed excellent wettability (three-phase contact angle = 89.88°). The results of the ζ-potentials and Fourier transform infrared spectroscopy demonstrate that electrostatic interaction forces and hydrogen bonding were the main forces involved in the formation of ZAHPs. On this basis, we prepared W1/O/W2 emulsions with other preparation parameters and observed their microstructures by optical microscopy and confocal laser scanning microscope. The multi-chambered structures of W1/O/W2 emulsions were successfully visualized. Finally, the W1/O/W2 emulsions were coated with PLL-alginate-Ca using the solution extrusion method. The results of the in vitro colonic digestion stage reveal that the survival rate of probiotics in the microgel beads was about 75.11%, which was significantly higher than that of the free. Moreover, probiotics encapsulated in microgel beads also showed positive storage stability. Apple pectin would serve as both an emulsifier and a prebiotic. Thus, the results indicate that the "egg-box" shaped microgel beads, designed on the basis of pH-sensitive and enzyme-triggered mechanisms, can enhance the efficiency of probiotics translocation in the digestive tract and mediate spatiotemporal controlled release.

Thermally reversible emulsion gels and high internal phase emulsions based solely on pea protein for 3D printing

This work compared thermally reversible emulsion gels and high internal phase emulsions (HIPEs) based on pea protein for 3D printing at different oil volume fraction (ϕ) ranging from 0.2 to 0.8. Thermally reversible emulsion gels were obtained with ϕ from 0.2 to 0.65, which melt into liquid emulsion as temperature increased, and recovered into emulsion gels upon cooling as reflected by consistent rheological measurements. The Fourier Transform Infrared Spectroscopy (FTIR) analysis revealed the transition between intermolecular β-sheet at 1630 cm−1 and intramolecular β-sheet at 1618 cm−1 through reversible H-bonds upon changes in temperature, which was responsible for the repeated thermo-reversibility. With ϕ increased to 0.74–0.8, HIPEs were confirmed by Scanning Electron Microscope (SEM) and Confocal Laser Scanning Microscope (CLSM), yet the thermo-reversibility was not maintained. In addition, the thermally reversible emulsion gels exhibited strong shear-thinning and thixotropic recovery through the untangling and reformation of the H-bonded gel network. Whereas deformation and restructuring of the close packed droplets were responsible for such behaviors in HIPEs. 3D printing test has further demonstrated excellent performance of thermally reversible emulsion-filled gel with ϕ 0.4, comparable to HIPE with ϕ 0.74 b y enabling printed objects with high shape fidelity (printability index (Pr) of 0.95–1.08) and shape stability (prints height maintenance (Hm) of 82.2%–87.8%). This research opens up thermally reversible emulsion gels solely from sustainable protein source for 3D printing, with low oil content and no synthetic ingredients needed.

Tumor microenvironment (Part I): Tissue integrity in a rat model of peripheral neural cancer

ICAM–1 (intercellular adhesion molecule 1) and MPZ (myelin protein zero) are thought to be a factor in the integrity of nerve tissues. In this report, we attempted to trace the expression of ICAM–1, responsible for cell-to-cell adhesion, and of MPZ, the main constituent of myelin sheath, in malignant tissues of the sciatic nerve (SN) in inbred male Copenhagen rats. AT–1 Cells (anaplastic tumor 1) were injected in the perineurial sheath, and tissues of the SNs were collected after 7, 14 and 21 days and compared to a sham-operated group of rats (n = 6 each). Tissues were sectioned and histologically examined, under light microscope, and stained for measuring the immunoreactivity of ICAM–1 and MPZ under laser scanning microscope. The cancer model was established, and the tumor growth was confirmed. ICAM–1 showed severe decreases, proportional to the growing anaplastic cells, as compared to the sham group. MPZ revealed, however, a distinct defensive pattern before substantially decreasing in a comparison with sham. These results support the notion that malignancies damage peripheral nerves and cause severe axonal injury and loss of neuronal integrity, and clearly define the role of ICAM–1 and MPZ in safeguarding the nerve tissues.

Design and synthesis of fluorescent BODIPY derivatives with D-A structure for cancer cell imaging

Boron-dipyrromethene (BODIPY) as a traditional fluorescent dye has received increasing attention because of its high molar extinction coefficient, easy modification, and good light stability. In this work, three BODIPY compounds (BDP-1, BDP-2 and BDP-3) were designed and synthesized by introducing PEG chains and a naphthalimide (NI) group onto the parent structure of BODIPY. Due to the electron-withdrawing properties of NI, the compounds of BDP-2 and BDP-3 formed a typical donor (D)-acceptor (A) structure, which led to the red-shifted absorption and fluorescence spectrum. The synthesized BODIPY exhibited intense green emission, with fluorescence quantum yield of 0.66, 0.29, and 0.19 for BDP-1, BDP-2 and BDP-3 respectively. The decrease in fluorescence of BDP-2 and BDP-3 was probably ascribed to the enhanced intramolecular charge transfer because of the introduction of the NI group. The good biocompatibility and low cytotoxicity of the synthesized BODIPY compounds were verified by the CCK-8 assay. The subcellular localization of the three compounds was further evaluated through a confocal laser scanning microscope. The results showed that these BODIPY derivatives were mainly localized in the lysosomes of cancer cells. Thus, the synthesized BODIPY fluorescent dyes show promising applications in tumor imaging benefiting from their good biocompatibility and strong fluorescence.

Effect of blood and artificial saliva contamination on marginal adaptation and sealing ability of different retrograde filling materials: A comparative analysis.

The objective of this study was to evaluate the effect of blood and artificial salivary contamination of different root-end filling materials on microleakage using a confocal laser scanning microscope and on marginal adaptation using a scanning electron microscope. Eighty noncarious single-rooted teeth with mature apices were taken. After retro-cavity preparation, they were randomly assigned into two major groups (n = 40). They were contaminated with blood and artificial saliva, respectively. Each major group was divided randomly into four subgroups (n = 10) and filled as follows: Subgroup A, Biodentine; Subgroup B, bioactive bone cement; Subgroup C, Cention N; and Subgroup D, Bio-C Repair. The samples were sectioned transversely at 1 and 2 mm from the root apex and checked under a confocal laser scanning microscope for microleakage and under an scanning electron microscope for marginal adaptation. The average mean values were calculated. Independent samples t-tests, paired t-tests, and one-way analysis of variance with Tukey's post hoc tests were done to analyze the data. All the tested materials showed marginal gaps and dye leakage. The Bio-C Repair group showed the least mean marginal gap and dye leakage values, followed by bioactive bone cement, Biodentine, and Cention N, respectively, in both blood and artificial saliva contamination. However, the mean marginal gaps and dye leakage between the major groups were statistically insignificant. In an overall comparison, Bio-C Repair was found to be superior in terms of marginal adaptation and sealing ability under the test conditions.

Comparison of penetration depth of chitosan, zinc oxide, and silica-doped titanium novel nanoparticle irrigant solutions – A confocal laser scanning microscopic in vitro study

ABSTRACT Aim: The aim of this study was to evaluate the penetration depth of three different irrigant solutions incorporated with nanoparticles (NPs) using a confocal laser scanning microscope. Methods: Forty-two single-rooted teeth were used. Access cavities were prepared, and working length was determined. The samples were split into three groups at random (n = 14). Group 1 – chitosan NPs, Group 2 – zinc oxide NPs, and Group 3 – silica-doped titanium NPs. Each group was further divided into subgroup A with activation and subgroup B without activation. Each tooth received final irrigants for 1 min. Each sample is sectioned at 3 mm from the apex. A confocal laser scanning microscope was used to observe the irrigant penetration. Results: The results showed that there was a significant difference in the penetration depth in Group 2A and Group 3A. Conclusion: Activation with PATS Vario increases the penetration when compared to conventional syringe, and zinc oxide NP solution showed increased penetration when compared with other groups.

Confocal laser scanning evaluation of the influence of ledges on root canal disinfection

AbstractThe aim was to analyse the influence of an apical ledge on root canal disinfection. Forty‐four single‐rooted teeth were micro‐CT scanned and inoculated with Enterococcus faecalis. In Group S shaping was performed with ProTaper Next (PTN) up to X3 at working length (WL). In Group L an apical ledge was created with K‐Files #40 and shaping completed up to PTN X3. NaOCl 5% and EDTA 10% irrigant solutions were alternated. Confocal laser scanning microscope (CLSM) and viability staining were used to analyse the proportions of dead (red) and live (green) bacteria and penetration ability inside dentinal tubules. Data were analysed with the Mann–Whitney test with Bonferroni correction (p &lt; 0.05). In Group L the amount of red fluorescence resulted significantly lower, and penetration ability was decreased in the apical and middle portion (p &lt; 0.05). The presence of an apical ledge may negatively influence the disinfection both in the apical and middle third.

Autofluorescence lifetime flow cytometry with time-correlated single photon counting.

Autofluorescence lifetime imaging microscopy (FLIM) is sensitive to metabolic changes in single cells based on changes in the protein-binding activities of the metabolic co-enzymes NAD(P)H. However, FLIM typically relies on time-correlated single-photon counting (TCSPC) detection electronics on laser-scanning microscopes, which are expensive, low-throughput, and require substantial post-processing time for cell segmentation and analysis. Here, we present a fluorescence lifetime-sensitive flow cytometer that offers the same TCSPC temporal resolution in a flow geometry, with low-cost single-photon excitation sources, a throughput of tens of cells per second, and real-time single-cell analysis. The system uses a 375 nm picosecond-pulsed diode laser operating at 50 MHz, alkali photomultiplier tubes, an FPGA-based time tagger, and can provide real-time phasor-based classification (i.e., gating) of flowing cells. A CMOS camera produces simultaneous brightfield images using far-red illumination. A second PMT provides two-color analysis. Cells are injected into the microfluidic channel using a syringe pump at 2-5 mm/s with nearly 5 ms integration time per cell, resulting in a light dose of 2.65 J/cm2 that is well below damage thresholds (25 J/cm2 at 375 nm). Our results show that cells remain viable after measurement, and the system is sensitive to autofluorescence lifetime changes in Jurkat T cells with metabolic perturbation (sodium cyanide), quiescent versus activated (CD3/CD28/CD2) primary human T cells, and quiescent versus activated primary adult mouse neural stem cells, consistent with prior studies using multiphoton FLIM. This TCSPC-based autofluorescence lifetime flow cytometer provides a valuable label-free method for real-time analysis of single-cell function and metabolism with higher throughput than laser-scanning microscopy systems.

Pickering emulsion gel of polyunsaturated fatty acid-rich oils stabilized by zein-tannic acid green nanoparticles for storage and oxidation stability enhancement

HypothesisPoor storage stability and oxidative deterioration are the common drawbacks of edible oils rich in polyunsaturated fatty acids (PUFAs). We hypothesized that the natural zein/tannic acid self-assembly nanoparticles (ZT NPs) could be employed as stabilizers to anchor at the oil–water interface, thus constructing Pickering emulsion gel (PKEG) system for three types of PUFA-rich oils, soybean oil (SO), fish oil (FO) and cod liver oil (CLO), to improve the storage and oxidation stability. ExperimentsZT NPs were prepared by the anti-solvent coprecipitation method, and the three-phase contact angle, FT-IR, and XRD were mainly characterized. To observe the shell-core structure and oil–water interface details of SO/FO/CLO PKEGs by confocal laser scanning microscope and cryo-scanning electron microscope. Accelerated oxidation of FO was performed to assess the protective effect of PKEG on lipids. FindingsThe SO, FO, and CLO PKEGs stabilized by 2 % ZT NPs, with oil fraction (φ = 0.5–0.6), were obtained. PKEGs show high viscoelasticity, clear shell-core structure spatial network structure, and ideal storage stability. Under accelerated oxidation, the degree of oxidative rancidity of FO PKEG was obviously lower than that of free FO. Overall, this work opens up new possibilities for using natural PKEG to prevent oxidative deterioration and prolong the shelf-life of PUFA-rich oils.

Thermal stabilization enhancement of diamond films via boron doping and its antioxidant mechanism

Diamond films possess numerous extraordinary physical and chemical properties, but their high-temperature applications are limited by its poor thermal stability. Herein, we fabricated the undoped microcrystalline diamond (MCD) film and heavy boron-doped diamond (h-BDD-1, 2.94 × 1021 atoms·cm−3; h-BDD-2, 9.10 × 1021 atoms·cm−3) films to investigate the effect of boron doping on the thermal stability of diamond films. A series of evidences, including air annealing experiments, thermogravimetric-differential scanning calorimetry (TG-DSC) analysis, and in-situ ultra-high-temperature confocal laser-scanning microscope (CLSM), demonstrated that the thermal stability of diamond films was obviously enhanced by boron doping. Consequently, the h-BDD-2 film exhibited the best thermal stability with an initial oxidation temperature above 900 °C, which was much higher than that of MCD film (709 °C). By deconvolving the X-ray photoelectron spectra, we believed that the transition of B–C bonds to stable oxy-boron carbide complexes (B4O–C, B3O–C) is the key to improving the thermal stability. These results not only reveal a new antioxidant mechanism of boron-doped diamond films but also pave a way for the application of diamond in high-temperature environments.

Diclofenac sodium effectively inhibits the biofilm formation of Staphylococcus epidermidis.

Staphylococcus epidermidis is an opportunistic pathogen commonly implicated in medical device-related infections. Its propensity to form biofilms not only leads to chronic infections but also exacerbates the issue of antibiotic resistance, necessitating high-dose antimicrobial treatments. In this study, we explored the use of diclofenac sodium, a non-steroidal anti-inflammatory drug, as an anti-biofilm agent against S. epidermidis. In this study, crystal violet staining and confocal laser scanning microscope analysis showed that diclofenac sodium, at subinhibitory concentration (0.4 mM), significantly inhibited biofilm formation in both methicillin-susceptible and methicillin-resistant S. epidermidis isolates. MTT assays demonstrated that 0.4 mM diclofenac sodium reduced the metabolic activity of biofilms by 25.21-49.01% compared to untreated controls. Additionally, the treatment of diclofenac sodium resulted in a significant decrease (56.01-65.67%) in initial bacterial adhesion, a crucial early phase of biofilm development. Notably, diclofenac sodium decreased the production of polysaccharide intercellular adhesin (PIA), a key component of the S. epidermidis biofilm matrix, in a dose-dependent manner. Real-time quantitative PCR analysis revealed that diclofenac sodium treatment downregulated biofilm-associated genes icaA, fnbA, and sigB and upregulated negative regulatory genes icaR and luxS, providing potential mechanistic insights. These findings indicate that diclofenac sodium inhibits S. epidermidis biofilm formation by affecting initial bacterial adhesion and the PIA synthesis. This underscores the potential of diclofenac sodium as a supplementary antimicrobial agent in combating staphylococcal biofilm-associated infections.