Microscopy Methods Research Articles

Understanding of the correlation of structural and microwave dielectric performance of (1-x)MgTiO3 - xCaTiO3 ceramics for dielectric resonator antenna applications

In this article, the effect of CaTiO3 on the structural and microwave dielectric characteristics of MgTiO3 has been discussed. The (1-x)MgTiO3 - xCaTiO3 (abbreviated as (1-x)MT- xCT) for x = 0.025, 0.05, 0.075, 0.1 ceramics have been developed using a solid-state reaction route. The structural as well as morphological characteristics of specimens have been observed by employing X-ray diffraction and scanning electron microscope methods. The Rietveld refinement technique of X-ray diffraction data have been conducted for the refinement of structural phases. It suggests that two crystallographic phases have been present in all materials. The SEM images display dense well-identified grains of (1-x)MT - xCT (for x = 0.025–0.1) ceramics. The Raman spectroscopy method has been employed to detect the symmetry of ceramics and modes of vibration of the materials. The microwave dielectric characteristics like dielectric permittivity (εr), loss (tanδ), quality factor (Q×f), and temperature coefficient at resonant frequency (τf) of (1-x)MT - xCT (for x = 0.025–0.1) ceramics have been thoroughly investigated. The bond strength, bond valency, bond energy, and tolerance factor of (1-x)MT - xCT (for x = 0.025–0.1) materials have been evaluated and their relation with microwave dielectric characteristics has been discussed. The (1-x)MT - xCT (for x = 0.025–0.1) ceramics show excellent dielectric properties with near to zero τf. In addition, DRAs have been designed with (1-x)MT - xCT (for x = 0.025–0.1) materials as dielectric resonators and various antenna properties have been studied with the help of HFSS software. This work suggests that (1-x)MT - xCT for (x=0.05) material can be a promising candidate for DRA applications working in the C band.

Al-Ni-Co decagonal quasicrystal application as an energy-effective catalyst for phenylacetylene hydrogenation

Intermetallic catalysts, including quasicrystals, are considered a sustainable alternative to noble metal-based catalysts in hydrogenation reactions. This study discusses the manufacturing and catalytic potential of an Al-Ni-Co quasicrystalline alloy. Energy-effective and simple catalyst production was provided by a melt-spinning process. The obtained ribbons, characterized in terms of microstructure, phase and chemical composition using X-ray diffraction, scanning and transmission microscopy methods, were composed of a decagonal quasicrystalline phase with traces of crystalline phases. The form of the melt-spun ribbons ensured easy application in the phenylacetylene hydrogenation reaction. The catalyst provided a substrate conversion of approximately 80% and a styrene selectivity of 54% after 1 h of reaction carried out under mild conditions. The repeatability of the reaction course was verified, with a maximum deviation of 10%. Moreover, the catalyst recovered after the reaction was evaluated in terms of its phase composition and surface changes. X-ray diffractograms confirmed the phase stability, however, the surface degradation and oxidation occurred. The catalytic activity after three months of catalyst storage is also discussed.

Ultrastructure of the mesonephros of whitefishes from the Lake Baikal basin.

The article presents a study of the mesonephros ultrastructure of Baikal omul Coregonus migratorius, Baikal whitefish Coregonus baicalensis, and a cross between Baikal whitefish and humpback whitefish (C. baicalensis × Coregonus pidschian). The mesonephros ultrastructure was studied using electron microscopy methods. The results of the study show that the number of mature granulocytes is a systematic feature and does not depend on the ecology of fish. The quantitative characteristics of blood cells and the ultrastructural features of leukocytes in the mesonephros are associated with the functioning of the nonspecific defence system in fish. Morphological diversity of epithelial cells in nephron tubules is the ancestral characteristic of the modern omul population, associated with geological and climatic events in the history of Lake Baikal. The development of haematopoietic tissue in the mesonephros, the ultrafine structure of ion-transporting interstitial cells, as well as some ultrastructural features found in the nephron, reflect the adaptive capabilities of the species to live in the ultra-deep Lake Baikal.

Microscopic methods to study meat and meat product quality

Reliable information about meat quality at all stages of the production process is necessary to ensure high quality of meat products. The structure of muscle, connective and fat tissues plays a direct role in formation of meat quality. Microscopic methods allow investigating the meat structure and determining its change depending on a range of endo- and exogenous factors (animal species, breed, sex, conditions of raising and slaughter) and on a type of technological processing. The paper presents the main directions of using microscopic analysis in investigation of meat and finished meat products. An advantage of microscopy is presentation of results in a visual form as well as a possibility of performing morphometry, including with the use of computer systems of image analysis, and obtaining quantitative characteristics of structures. Most common are light microscopy and electron microscopy. Due to various staining procedures, light microscopy enables detecting different components of a sample, studying topography and morphology of tissues and cells. Electron microscopy gives information about the ultrastructure of cells and their chemical composition. The paper discusses possibilities of microscopy in assessment of composition and detection of falsification of finished meat products. It has been noted that the use of several approaches and methods of staining allows reliable identification of many components, including components of plant origin. Histological methods can ensure detection of falsification and control of meat product composition at the state level.

Application of Weak-Beam Dark-Field STEM for Dislocation Loop Analysis†.

Nanoscale dislocation loops formed by irradiation can significantly contribute to both irradiation hardening and embrittlement of materials when subjected to extreme nuclear reactor environments. This study explores the application of weak-beam dark-field (WBDF) scanning transmission electron microscopy (STEM) methods for quantitative irradiation-induced defect analysis in crystalline materials, with a specific focus on dislocation loop imaging and analysis. A high-purity Fe-5 wt% Cr model alloy was irradiated with 8 MeV Fe2+ ions at 450°C to a fluence of 8.8 × 1019 m-2, inducing dislocation loops for analysis. While transmission electron microscopy (TEM) has traditionally been the primary tool for dislocation imaging, recent advancements in STEM technology have reignited interest in using STEM for defect imaging. This study introduces and compares three WBDF STEM methods, demonstrating their effectiveness in suppressing background contrasts, isolating defect information for dislocation loop type classification, providing finer dislocation line images for small loop analysis, and presenting inside-outside contrast for identifying loop nature. Experimental findings indicate that WBDF STEM methods surpass traditional TEM approaches, yielding clearer and more detailed images of dislocation loops. The study concludes by discussing the potential applications of WBDF STEM techniques in defect analysis, emphasizing their adaptability across various material systems beyond nuclear materials.

Live Cell Protein Imaging of Tandem Complemented-GFP11-Tagged Coiled-Coil Domain-Containing Protein-124 Identifies this Factor in G3BP1-Induced Stress-Granules.

Coiled-coil domain-containing 124 protein is a multifunctional RNA-binding factor, and it was previously reported to interact with various biomolecular complexes localized at diverse subcellular locations, such as the ribosome, centrosome, midbody, and nucleoli. We aimed to better characterize the subcellular CCDC124 translocation by labelling this protein with a fluorescent tag, followed by laser scanning confocal microscopy methods. As traditional GFP-tagging of small proteins such as CCDC124 often faces limitations like potential structural perturbations of labeled proteins, and interference of the fluorescent-tag with their endogenous cellular functions, we aimed to label CCDC124 with the smallest possible split-GFP associated protein-tagging system (GFP11/GFP1-10) for better characterization of its subcellular localizations and its translocation dynamics. By recombinant DNA techniques we generated CCDC124-constructs labelled with either single of four tandem copies of GFP11 (GFP11 × 1::CCDC124, GFP11 × 4::CCDC124, or CCDC124::GFP11 × 4). We then cotransfected U2OS cells with these split-GFP constructs (GFP11 × 1(or X4)::CCDC124/GFP1-10) and analyzed subcellular localization of CCDC124 protein by laser scanning confocal microscopy. Tagging CCDC124 with four tandem copies of a 16-amino acid short GFP-derived peptide-tag (GFP11 × 4::CCDC124) allowed better characterization of the subcellular localization of CCDC124 protein in our model human bone osteosarcoma (U2OS) cells. Thus, by this novel methodology we successfully identified GFP11 × 4::CCDC124 molecules in G3BP1-overexpression induced stress-granules by live cell protein imaging for the first time. Our findings propose CCDC124 as a novel component of the stress granule which is a membraneless organelle involved in translational shut-down in response to cellular stress.

From Pixels to Information: Artificial Intelligence in Fluorescence Microscopy

This review explores how artificial intelligence (AI) is transforming fluorescence microscopy, providing an overview of its fundamental principles and recent advancements. The roles of AI in improving image quality and introducing new imaging modalities are discussed, offering a comprehensive perspective on these changes. Additionally, a unified framework is introduced for comprehending AI‐driven microscopy methodologies and categorizing them into linear inverse problem‐solving, denoising, and nonlinear prediction. Furthermore, the potential of self‐supervised learning techniques that address the challenges associated with training the networks are explored, utilizing unlabeled microscopy data to enhance data quality and expand imaging capabilities. It is worth noting that while the specific examples and advancements discussed in this review focus on fluorescence microscopy, the general approaches and theories are directly applicable to other optical microscopy methods.

Changes of wood surfaces treated with natural-based products – Structural and properties investigation

Preservative systems based on vegetable seed oils and natural waxes from renewable sources may confer protection to wood under exposure to various environmental conditions. These, as non-toxic substances, can form an environmentally friendly and efficient protective layer on the wood surfaces, with beneficial effects on their water resistance and dimensional stability. Thus, these natural coatings may hinder biodegradation of wood products to a certain degree. In present paper, softwood samples (from Abies alba fir tree species), prepared as dried discs (25 to 30 mm diameter, 8 to 10 mm thickness), were surface impregnated by dipping using vegetable oils, namely Asclepias syriaca seed oil, and soybean oil, respectively. Beeswax treatment was also applied for comparison purposes. Surface chemistry and morphology, biodegradation process under controlled and simulated natural conditions, and water sorption behavior of wood samples were investigated. Fourier Transform Infrared spectroscopy, X-ray diffraction, and scanning electron microscopy methods were used for investigation of surface changes in wood samples before and after impregnation with natural based products, as well as under biodegradation conditions in soil burial tests.

Direct observation on the phase separation of polymer membrane embryos confined by microfluidics chips

In this paper, the growth of finger-like droplets inside various polymer membrane embryos induced by non-solvent phase separation (NIPS) is studied in details by using a standardized optical microscopic observation method. The boundary conditions of the phase separation are well-defined by microfluidics technology. The growth of finger-like droplets and Marangoni convection inside them are tracked and recorded by high-speed camera of sharp resolutions. Effects of the geometry of the membrane embryos (thickness/shape/size), humidity of the atmosphere and the viscosity of the embryos on the growth of finger-like droplets are systematically studied. Except the qualitative observations that are consistent to previous relative reports, more quantitative data are provided in this paper. The diffusing speed of the interface of the developing large droplets is measured by template matching algorithm. Strikingly, the time-dependent diffusing speed of the developing large droplets shows a wave-like declining pattern, which indicates that fluctuating boundary conditions are surrounding the membrane embryos. The Reynolds number of the flow inside the developing large droplets is estimated at the order of 10−5, the magnitude of which is 2 order smaller than that of a swimming sperm.

Study on mechanical properties and meso-damage mechanism of carbon-polyvinyl alcohol hybrid fiber reinforced recycled coarse aggregate concrete under the coupling action of sodium sulfate and dry-wet cycles

In order to investigate the influence of the coupling effect of sodium sulfate dry-wet (DW) cycles on the mechanical properties of carbon-polyvinyl alcohol hybrid fiber reinforced recycled coarse aggregate concrete (HFRAC), HFRAC specimens were subjected to uniaxial compression tests. The total volume fraction of fibers was kept constant at 0.4%, and various volume fractions of carbon fibers (CF) were added to the recycled coarse aggregate concrete (RAC), namely 0%, 0.1%, 0.2%, 0.3%, and 0.4%, with polyvinyl alcohol (PVA) fibers replacing a portion of CF in equivalent volume. Uniaxial compressive stress–strain curves were obtained for HFRAC specimens subjected to different numbers of DW cycles (0, 30, 60, and 120 cycles). Microscopic testing methods including acoustic emission (AE), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), X-ray diffraction (XRD), and nuclear magnetic resonance (NMR) were employed to analyze the microstructure, erosion product composition, and pore structure changes of the specimens. The results revealed that, for RAC specimens, the peak stress (σp) and elastic modulus (E) initially increased and then decreased, whereas the peak strain (εp) initially decreased and then increased with an increase in the number of DW cycles. After adding hybrid fibers, the σp values of CF, Hybrid 3:1, Hybrid 1:1, Hybrid 1:3, and PVA specimens increased (or decreased) by 12.0%, 11.43%, 0.9%, −9.5%, and −8.5%, respectively, compared to RAC. The Hybrid 1:3 specimen exhibited a negative hybrid effect. With an increase in the erosion age, the Hybrid 3:1 specimen demonstrated enhanced resistance to sulfate attack and achieved the highest σp value after 120 cycles. The influence of sulfate DW cycles coupling on the microscopic damage mechanism of HFRAC was analyzed using a statistical damage constitutive model. The microscopic damage evolution process was characterized by five characteristic parameters: E0, εa, εh, εb, and H. The analysis results indicated a clear correlation between the variation of these characteristic parameters and the number of DW cycles. An effective connection was established between the microscopic damage mechanism and the macroscopic nonlinear constitutive behavior. The phenomenon of relative lagging of acoustic emission signals during the uniaxial compression process was explained from the perspective of effective stress. This study provides a theoretical basis for the promotion and application of RAC under sulfate DW cycle environments.

The effect of temperature and ultrasonic power on the microstructure evolution of coal modified by clean fracturing fluid: An experimental study

Ultrasonic-assisted fracturing permeability enhancement technology offers several advantages, including a broad range of hydraulic fracturing permeability enhancement, microstructure modified by fracturing fluid, and ultrasonic promotion of gas desorption and extraction. This technology can significantly improve the efficiency of coalbed methane extraction and has broad application prospects. To investigate the effect of ultrasonic action power and temperature on the microstructure evolution of coal by using clean fracturing fluid, experiments were conducted using coal samples from a mine in Shenyang. The pore structure of these samples was analyzed by scanning electron microscopy (SEM) and mercury intrusion method (MIP), while the chemical microstructure was evaluated with energy dispersive spectroscopy (EDS), X-ray diffraction experiment (XRD) and Fourier transform infrared spectroscopy (FTIR). The results showed that ultrasonic treatment improved the pore structure characteristics and connectivity by facilitating chemical reactions between fracturing fluid and mineral impurities. Specifically, when the ultrasonic parameter was set at 600W-60 °C, the pore volume of coal samples increased by 26.09 %, tortuosity decreased by 31.81 %, pore fractal dimension increased by 3.93 %, permeability increased by 100.45 %, aromaticity increased by 16.69 %, and the degree of branching aliphatic side-chains increased by 108.70 %. Under the same ultrasonic power, the effective temperature for ultrasonic-assisted fracturing fluid treatment on coal rock was found to be 60 °C. Additionally, under the same temperature conditions, an increase in ultrasonic power resulted in significant enhancements in both pore structure parameters and chemical structure parameters in coal, showing a certain linear relationship. The research results enrich the theoretical system of coalbed methane mining and provide theoretical guidance for the field practice of this technology.

Investigating Retinal Circuits and Molecular Localization by Pre-Embedding Immunoelectron Microscopy.

The retina comprises numerous cells forming diverse neuronal circuits, which constitute the first stage of the visual pathway. Each circuit is characterized by unique features and distinct neurotransmitters, determining its role and functional significance. Given the intricate cell types within its structure, the complexity of neuronal circuits in the retina poses challenges for exploration. To better investigate retinal circuits and cross-talk, such as the link between cone and rod pathways, and precise molecular localization (neurotransmitters or neuropeptides), such as the presence of substance P-like immunoreactivity in the mouse retina, we employed a pre-embedding immunoelectron microscopy (immuno-EM) method to explore synaptic connections and organization. This approach enables us to pinpoint specific intercellular synaptic connections and precise molecular localization and could play a guiding role in exploring its function. This article describes the protocol, reagents used, and detailed steps, including (1) retina fixation preparation, (2) pre-embedding immunostaining, and (3) post-fixation and embedding.

Laser synthesis of oxide nanoparticles with controlled gas condensation

In this work, the oxide nanopowders of Al2O3, ZrO2, Y2O3, Gd2O3, CeO2, and SiO2 were synthesized by CW CO2 laser vaporization technique with controlled gas condensation in an inert atmosphere. Methods for controlling the size of the resulting nanoparticles by adjusting the gas composition and pressure during the vaporization process have been demonstrated. The potential for producing ultrasmall oxide nanoparticles with dimensions less than 5 nm has been shown. The size distribution of nanoparticles taken from different parts of the evaporation-condensation tract was studied using scanning (SEM) and transmission (TEM) electron microscopy methods. The effect of synthesis conditions (pressure and composition of the inert gas) on characteristics of the nanoparticles is discussed. Using a wide class of simple oxides as the example, it is shown that the powders synthesized by the laser method consist of three types of particles: target spherical particles with a diameter of 3–20 nm (more than 98%), larger spherical particles with a diameter of 50–200 nm, and shapeless large particles with sizes more than 200 nm. The possibility of separating large particles from the main particles using the original labyrinth system for gas pumping is shown. The obtained particles with controlled sizes can be effectively used in various applications, in particular, for the preparation of catalysts and adsorbents.

A new diagnostic technique for identifying Angiostrongylus spp. larvae in intermediate snail species by examining the buccal cavity

BackgroundAngiostrongyliasis is a zoonotic parasitic disease caused by the rat lungworm Angiostrongylus cantonensis. The intermediate hosts of A. cantonensis are gastropods, and snail species such as Pomacea canaliculata play a key role in the transmission of human angiostrongyliasis. Detecting A. cantonensis infection in snails is an important component of epidemiological surveillance and the control of angiostrongyliasis.MethodsIn this study, a new method for diagnosing A. cantonensis infection in gastropods was developed by recovering larvae from the buccal cavity of three snail species. The entire buccal cavity of a snail was extracted, and the tissue was pressed between two microscope slides to observe whether A. cantonensis larvae were present. Our new method was compared with traditional pathogenic detection methods of lung microscopy, tissue homogenization, and artificial digestion. We artificially infected 160 P. canaliculata, 160 Cipangopaludina chinensis, and 160 Bellamya aeruginosa snails with A. cantonensis. Then, the four different detection methods were used to diagnose infection in each snail species at 7, 14, 21, and 28 days post exposure.ResultsWe found no significant difference in the percentages of infected P. canaliculata snails using the four methods to detect A. cantonensis larvae. The radula pressing method had a mean detection rate of 80%, while the lung microscopy (81.3%), tissue homogenization (83.8%), and artificial digestion (85%) methods had slightly greater detection rates. Similarly, the percentages of infected C. chinensis snails that were detected using the radula pressing (80%), tissue homogenization (82.1%), and artificial digestion (83.8%) methods were not significantly different. Finally, the percentages of infected B. aeruginosa snails that were detected using the radula pressing (81.3%), tissue homogenization (81.9%), and artificial digestion (81.4%) methods were not significantly different. These results showed that the radula pressing method had a similar detection rate to traditional lung microscopy, tissue homogenization, or artificial digestion methods.ConclusionsThis study demonstrates a new method for the qualitative screening of gastropods that act as intermediate hosts of A. cantonensis (and other Angiostrongylus species), provides technical support for the control of human angiostrongyliasis, and furthers research on A. cantonensis.Graphical

Hypoxic extracellular vesicles from hiPSCs protect cardiomyocytes from oxidative damage by transferring antioxidant proteins and enhancing Akt/Erk/NRF2 signaling

BackgroundStem cell-derived extracellular vesicles (EVs) are an emerging class of therapeutics with excellent biocompatibility, bioactivity and pro-regenerative capacity. One of the potential targets for EV-based medicines are cardiovascular diseases (CVD). In this work we used EVs derived from human induced pluripotent stem cells (hiPSCs; hiPS-EVs) cultured under different oxygen concentrations (21, 5 and 3% O2) to dissect the molecular mechanisms responsible for cardioprotection.MethodsEVs were isolated by ultrafiltration combined with size exclusion chromatography (UF + SEC), followed by characterization by nanoparticle tracking analysis, atomic force microscopy (AFM) and Western blot methods. Liquid chromatography and tandem mass spectrometry coupled with bioinformatic analyses were used to identify differentially enriched proteins in various oxygen conditions. We directly compared the cardioprotective effects of these EVs in an oxygen-glucose deprivation/reoxygenation (OGD/R) model of cardiomyocyte (CM) injury. Using advanced molecular biology, fluorescence microscopy, atomic force spectroscopy and bioinformatics techniques, we investigated intracellular signaling pathways involved in the regulation of cell survival, apoptosis and antioxidant response. The direct effect of EVs on NRF2-regulated signaling was evaluated in CMs following NRF2 inhibition with ML385.ResultsWe demonstrate that hiPS-EVs derived from physiological hypoxia at 5% O2 (EV-H5) exert enhanced cytoprotective function towards damaged CMs compared to EVs derived from other tested oxygen conditions (normoxia; EV-N and hypoxia 3% O2; EV-H3). This resulted from higher phosphorylation rates of Akt kinase in the recipient cells after transfer, modulation of AMPK activity and reduced apoptosis. Furthermore, we provide direct evidence for improved calcium signaling and sustained contractility in CMs treated with EV-H5 using AFM measurements. Mechanistically, our mass spectrometry and bioinformatics analyses revealed differentially enriched proteins in EV-H5 associated with the antioxidant pathway regulated by NRF2. In this regard, EV-H5 increased the nuclear translocation of NRF2 protein and enhanced its transcription in CMs upon OGD/R. In contrast, inhibition of NRF2 with ML385 abolished the protective effect of EVs on CMs.ConclusionsIn this work, we demonstrate a superior cardioprotective function of EV-H5 compared to EV-N and EV-H3. Such EVs were most effective in restoring redox balance in stressed CMs, preserving their contractile function and preventing cell death. Our data support the potential use of hiPS-EVs derived from physiological hypoxia, as cell-free therapeutics with regenerative properties for the treatment of cardiac diseases.

Exogenous Sodium Nitroprusside Affects the Redox System of Wheat Roots Differentially Regulating the Activity of Antioxidant Enzymes under Short-Time Osmotic Stress.

Nitric oxide (NO) is a multifunctional signalling molecule involved in the regulation of plant ontogenesis and adaptation to different adverse environmental factors, in particular to osmotic stress. Understanding NO-induced plant protection is important for the improvement of plant stress tolerance and crop productivity under global climate changes. The root system is crucial for plant survival in a changeable environment. Damages that it experiences under water deficit conditions during the initial developmental periods seriously affect the viability of the plants. This work was devoted to the comparative analysis of the pretreatment of wheat seedlings through the root system with NO donor sodium nitroprusside (SNP) for 24 h on various parameters of redox homeostasis under exposure to osmotic stress (PEG 6000, 12%) over 0.5-24 h. The active and exhausted solutions of SNP, termed as (SNP/+NO) and (SNP/-NO), respectively, were used in this work at a concentration of 2 × 10-4 M. Using biochemistry and light microscopy methods, it has been revealed that osmotic stress caused oxidative damages and the disruption of membrane cell structures in wheat roots. PEG exposure increased the production of superoxide (O2•-), hydrogen peroxide (H2O2), malondialdehyde (MDA), and the levels of electrolyte leakage (EL) and lipid peroxidation (LPO). Stress treatment enhanced the activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), the excretion of proline, and the rate of cell death and inhibited their division. Pretreatment with (SNP/+NO) decreased PEG-induced root damages by differently regulating the antioxidant enzymes under stress conditions. Thus, (SNP/+NO) pretreatment led to SOD, APX, and CAT inhibition during the first 4 h of stress and stimulated their activity after 24 h of PEG exposure when compared to SNP-untreated or (SNP/-NO)-pretreated and stress-subjected plants. Osmotic stress triggered the intense excretion of proline by roots into the external medium. Pretreatment with (SNP/+NO) in contrast with (SNP/-NO) additionally increased stress-induced proline excretion. Our results indicate that NO is able to mitigate the destructive effects of osmotic stress on the roots of wheat seedlings. However, the mechanisms of NO protective action may be different at certain periods of stress exposure.

Adaptive optics in an oblique plane microscope.

Adaptive optics (AO) can restore diffraction-limited performance when imaging beyond superficial cell layers in vivo and in vitro, and as such, is of interest for advanced 3D microscopy methods such as light-sheet fluorescence microscopy (LSFM). In a typical LSFM system, the illumination and detection paths are separate and subject to different optical aberrations. To achieve optimal microscope performance, it is necessary to sense and correct these aberrations in both light paths, resulting in a complex microscope system. Here, we show that in an oblique plane microscope (OPM), a type of LSFM with a single primary objective lens, the same deformable mirror can correct both illumination and fluorescence detection. Besides reducing the complexity, we show that AO in OPM also restores the relative alignment of the light-sheet and focal plane, and that a projection imaging mode can stabilize and improve the wavefront correction in a sensorless AO format. We demonstrate OPM with AO on fluorescent nanospheres and by imaging the vasculature and cancer cells in zebrafish embryos embedded in a glass capillary, restoring diffraction limited resolution and improving the signal strength twofold.

Low-Frequency Coherent Raman Imaging Robust to Optical Scattering.

We demonstrate low-frequency interferometric impulsive stimulated Raman scattering (ISRS) imaging with high robustness to distortions by optical scattering. ISRS is a pump-probe coherent Raman spectroscopy that can capture Raman vibrational spectra. Recording of ISRS spectra requires isolation of a probe pulse from the pump pulse. While this separation is simple in nonscattering specimens, such as liquids, scattering leads to significant pump pulse contamination and prevents the extraction of a Raman spectrum. We introduce a robust method for ISRS microscopy that works in complex scattering samples. High signal-to-noise ISRS spectra are obtained even when the pump and probe pulses pass through many scattering layers.

Examining the Synergic Effect of Exosomes Derived from Mouse Mesenchymal Stem Cells and Low-frequency Electromagnetic Field on Chondrogenic Differentiation

Background: Cartilage has intrinsically limited healing power, and regeneration of cartilage damages has remained a challenge. Secreted products of mesenchymal stem cells have shown a new therapeutic strategies for cartilage injuries. Also it has been observed that low frequency electromagnetic field plays a key role in biological processes. Objective: This research was performed to investigate the synergic effect of mesenchymal stem cell-derived exosomes and low frequency electromagnetic field on chondrogenic differentiation. Methods: In this in vitro study, mesenchymal stem cell-derived exosomes were identified using AFM, SEM, TEM microscopy, and DLS method. Cells were treated in chondrogenic medium by exosomes, low frequency electromagnetic field, and the synergy of both. The cell survival was examined using MTT and Annexin methods, and cartilage differentiation was confirmed by Alcian blue staining. The expression of Sox-9, Acan, Col 2a1 and Col 10a1 genes was examined via Real- Time PCR technique on day 14 post-treatment. Results: The results confirmed the presence of exosomes with an approximate size of less than 100 nm. The results of Alcian blue revealed greater expression of glycosaminoglycans in the synergic treatment group compared to the other groups. Real-time PCR showed a significant increase in the expression of Sox-9, Acan, and Col 2a1 genes, as well as a significant reduction of Col 10a1 gene expression in the synergic treatment group compared to other groups. Conclusion: This study indicated that the synergic effect of exosome and low-frequency electromagnetic fields would lead to enhanced chondrogenic differentiation, which can be further explored in future clinical studies

Application of digital image correlation for in-situ deformation studies using transmission electron microscopy

Digital Image correlation (DIC) is routinely applied using optical and scanning electron microscopy methods to map local strains during deformation. In this letter, we report its use for in-situ TEM imaged deformation evolution for a nanocrystalline metal. Since transmitted electrons give rise to complex contrast conditions, with such contrast associated with the mechanisms of deformation, one can track these changes if particular care is done in image analysis and interpretation. We describe these methodologies to enable this correlation through frame averaging. Using these collective correlation conditions, we found that the average DIC strain matches well with the far field strain values yielding increased confidence in its application. Through the utilization of DIC in the TEM, nanoscale deformation mechanisms can be further elucidated because of the increased spatial resolution offered by TEM.