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An optical test to unveil twisting of birefringent crystals in spherulites
摘要： Helical conformations and structures are frequently observed in materials. The presence of helices at points of the unit cell of a crystal, on a larger size scale in the crystalline lattice or even in the microscopic structure of crystals, affects the chemico-physical properties of a solid and, hence, also interactions with light. Here, attention has been drawn to the geometrical properties of helices produced by a hypothetical torque of a transparent crystal, and optical properties of twisted crystals easily observed by a polarizing microscope have been discussed. Radially grown spherulites are obtained by most substances crystallized from melt. The circular optical arrangement of behaviour of larger dimensions of spherulites, allows investigations otherwise hardly feasible on separate crystals. According to the torsional analysis of elongated bodies and the birefringence theory, information on the existence of helically shaped crystals can be deduced, as hereinafter explained, from the microscopic appearance and birefringence pattern of spherulites. Indeed, twisting decreases the birefringence throughout an elongated crystal and, therefore, also the birefringence of spherulites formed by twisted radial crystals is reduced.
关键词： optical microscopy,microstructure,helical deformation,birefringence,crystals,banding
更新于2025-10-24 16:38:51
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Vibrational spectroscopic imaging and live cell video microscopy for studying differentiation of primary human alveolar epithelial cells
摘要： Alveolar type II(ATII) cells in the peripheral human lung spontaneously differentiate towards alveolar type I(ATI) cells, thus enabling air-blood barrier formation. Here, linear Raman and coherent anti-Stokes Raman scattering (CARS) microscopy are applied to study cell differentiation of freshly isolated ATII cells. The Raman spectra can successfully be correlated to gradual morphological and molecular changes during cell differentiation. Alveolar surfactant rich vesicles in ATII cells are identified based on phospholipid vibrations, while ATI-like cells are characterized by the absence of vesicular structures. Complementary, CARS microscopy allows for 3D visualization of lipid vesicles within ATII cells and their secretion, while hyperspectral CARS enables the distinction between cellular proteins and lipids according to their vibrational signatures. This study paves the path for further label-free investigations of lung cells and the role of the pulmonary surfactant, thus also providing a basis for rational development of future lung therapeutics.
关键词： confocal laser scanning microscopy,confocal Raman microscopy,pneumocyte type II differentiation,CARS microscopy,cell imaging
更新于2025-09-23 15:23:52
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Multiphoton imaging for morphometry of the sandwich-beam structure of the human stapedial annular ligament
摘要： Background: The annular ligament of the human stapes constitutes a compliant connection between the stapes footplate and the peripheral cochlear wall at the oval window. The cross section of the human annular ligament is characterized by a three-layered structure, which resembles a sandwich-shaped composite structure. As accurate and precise descriptions of the middle-ear behavior are constrained by lack of information on the complex geometry of the annular ligament, this study aims to obtain comprehensive geometrical data of the annular ligament via multiphoton imaging. Methods: The region of interest containing the stapes and annular ligament was harvested from a fresh-frozen human temporal bone of a 46-years old female. Multiphoton imaging of the unstained sample was performed by detecting the second-harmonic generation of collagen and the autofluorescence of elastin, which are constituents of the annular ligament. The multiphoton scans were conducted on the middle-ear side and cochlear side of the annular ligament to obtain accurate images of the face layers on both sides. The face layers of the annular ligament were manually segmented on both multiphoton scans, and then registered to high-resolution mCT images. Results: Multiphoton scans of the annular ligament revealed 1) relatively large thickness of the core layer compared to the face layers, 2) asymmetric geometry of the face layers between the middle-ear side and cochlear side, and variation of their thickness and width along the footplate boundary, 3) divergent relative alignment of the two face layers, and 4) different fiber composition of the face layers along the boundary with a collagen-reinforcement near the anterior pole on the middle-ear side. Conclusion and outlook: Multiphoton microscopy is a feasible approach to obtain the detailed three-dimensional features of the human stapedial annular ligament along its full boundary. The detailed description of the sandwich-shaped structures of the annular ligament is expected to contribute to modeling of the human middle ear for precise simulation of middle-ear behavior. Further, established methodology in this study may be applicable to imaging of other middle-ear structures.
关键词： Stapes,Two-photon microscopy,Multiphoton microscopy,Core layer,Face layer,Annular ligament
更新于2025-09-23 15:23:52
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Successful optimization of reconstruction parameters in structured illumination microscopy – A practical guide
摘要： The impact of the different reconstruction parameters in super-resolution structured illumination microscopy (SIM) on image artifacts is carefully analyzed. These parameters comprise the Wiener filter parameter, an apodization function, zero-frequency suppression and modifications of the optical transfer function. A detailed investigation of the reconstructed image spectrum is concluded to be suitable for identifying artifacts. For this purpose, two samples, an artificial test slide and a more realistic biological system, were used to characterize the artifact classes and their correlation with the image spectra as well as the reconstruction parameters. In addition, a guideline for efficient parameter optimization is suggested and the implementation of the parameters in selected up-to-date processing packages (proprietary and open-source) is depicted.
关键词： Super resolution,Fluorescence microscopy,Structured illumination microscopy,Parameter estimation,Image reconstruction
更新于2025-09-23 15:23:52
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Slide-free imaging of hematoxylin-eosin stained whole-mount tissues using combined third-harmonic generation and three-photon fluorescence microscopy
摘要： Intraoperative margin assessment of surgical tissues during cancer surgery is clinically important, especially in the case of tissue conserving surgery like Mohs micrographic surgery in which minimization of the surgical area is considered crucial. Frozen pathology is the gold standard of assessing excised tissues for signs of remaining cancerous lesions. The current protocol, however, is time-consuming and labor-intensive. Instead of the complex frozen sectioning, staining, and traditional white light microscopy imaging protocol, optically-sectioned histopathological imaging of hematoxylin-eosin stained whole-mount skin tissues with a sub-femtoliter resolution is demonstrated by using nonlinear microscopy in this study. With our proposed method, the reagents of staining and the contrast of imaging are fully consistent with the current clinical standard of frozen pathology, thus facilitating rapid intraoperative assessment of surgical tissues for future applications.
关键词： hematoxylin-eosin,three photon microscopy,third harmonic generation microscopy,surgical border,margin assessment
更新于2025-09-23 15:23:52
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Disentangling topographic contributions to near-field scanning microwave microscopy images
摘要： We develop empirical models to predict the contribution of topographic variations in a sample to near-field scanning probe microwave microscopy (NSMM) images. In particular, we focus on |S11| images of a thin Perovskite photovoltaic material and a GaN nanowire. The difference between the measured NSMM image and this prediction is our estimate of the contribution of material property variations to the measured image. Prediction model parameters are determined from either a reference sample that is nearly free of material property variations or directly from the sample of interest. The parameters of the prediction model are determined by robust linear regression so as to minimize the effect of material property variations on results. For the case where the parameters are determined from the reference sample, the prediction is adjusted to account for instrument drift effects. Our statistical approach is fully empirical and thus complementary to current approaches based on physical models that are often overly simplistic.
关键词： Near-field scanning probe microwave microscopy,Signal extraction,GaN nanowire,Statistical methods,Perovskite materials,Atomic force microscopy
更新于2025-09-23 15:23:52
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Asymmetric strain: A critical role in domain evolution in (111) - Oriented ferroelectric films
摘要： Domain behavior of (111)- oriented perovskite ferroelectric films is significantly different from (001)-/(101)- oriented ones, resulting in enhancing property responses such as a superior susceptibility and a reduced coercive field. However, the domain structures and evolutions with the thickness of (111)-oriented ferroelectric films, which are crucial to further understand the distinctive properties, are still obscure. In this study, the ferroelectric domains of (111)- oriented PbTiO3 films are investigated by transmission electronic microscopy (TEM) and piezoresponse force microscopy (PFM). We identify the domain evolution with the film thicknesses under anisotropic strains imposed by the orthorhombic GdScO3 (101)O substrates. Contrast analysis and electron diffraction patterns reveal that only four ferroelectric variants evolve in PTO films: d2+, d2-, d3+ and d3-, with the polarization directions along [010], [010], [001] and [001], respectively. Two kinds of domain walls are formed: 'inclined' (011) domain walls for d2-/d3+ (d2+/d3-) domains, 'normal' (011) domain walls for d2+/d3- (d2-/d3+) domains. The width of periodically distributed d2+/d3+ (d2-/d3-) domains increases with film thickness following the square root rule. Aberration-corrected scanning transmission electronic microscopy demonstrates the lattice characteristics of domains in (111)- oriented PbTiO3 films are consistent with tetragonal ferroelectric domains. PFM studies reveal that both the out-of-plane and in-plane polarization components of d2-/d3+ (d2+/d3-) domains are non-identical, whereas the d2+/d3+ (d2-/d3-) domains possess uniform out-of-plane polarization component and non-uniform in-plane polarization component. This study discloses the domain structure in (111)-oriented tetragonal ferroelectric films under anisotropic strains and the association with the ferroelectric properties.
关键词： PbTiO3,Transmission electron microscopy,Ferroelectric,T,Perovskites,Piezoresponse force microscopy
更新于2025-09-23 15:23:52
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Superdomain structure and high conductivity at the vertices in the (111)-oriented epitaxial tetragonal Pb(Zr,Ti)O3 thin film
摘要： Recently, in ferroelectric materials, there have been many experimental efforts to find out more intriguing topological objects and their functionalities, such as conduction property. Here we investigated ferroelectric domain structures and related topological defects in the (111)-oriented epitaxial tetragonal PbZr0.35Ti0.65O3 thin film. Systematic piezoresponse force microscopy measurements revealed that the field-induced polarization switching can form thermodynamically stable superdomain structures composed of nano-sized stripe sub-domains. Within such superdomain structures, we observed the exotic equilateral triangular in-plane flux-closure domains composed of three stripe domain bundles with 120/120/120 degrees of separation. The conductive-atomic force microscopy measurements under vacuum showed that some vertices have significantly higher conductivity compared to other surrounding regions. This work highlights electric field-driven polarization switching and unique crystallographic symmetry (here, three-fold rotational symmetry) can generate exotic ferroelectric domain structures and functional topological defects, such as conductive vertices.
关键词： Vertex,Ferroelectric,Superdomain,Flux-closure domain,Piezoresponse force microscopy,Conductive-atomic force microscopy
更新于2025-09-23 15:23:52
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[IEEE 2018 IEEE International Symposium on the Physical and Failure Analysis of Integrated Circuits (IPFA) - Singapore (2018.7.16-2018.7.19)] 2018 IEEE International Symposium on the Physical and Failure Analysis of Integrated Circuits (IPFA) - Characterization of Multilayered Ceramic Capacitors via Piezoelectric Force Microscopy
摘要： The coupling between an electrical and mechanical response in a material is a fundamental property that provides functionality to a variety of applications ranging from sensors and actuators to energy harvesting and biology. Most materials exhibit electromechanical coupling in nanometer-sized domains. Therefore, to understand the relationship between structure and function of these materials, characterization on the nanoscale is required. This property can be directly measured in a non-destructive manner using piezoelectric force microscopy (PFM), a mode that comes standard in all atomic force microscopes (AFMs) from Park Systems. Additionally, PFM can be used as a spectroscopic tool to evaluate switching of piezoelectric domains. Here we demonstrate the utility of PFM for failure analysis of a multilayered ceramic capacitor. Correlative imaging of topography and electrical signals revealed discontinuous structures in the device that likely had a direct effect on device performance. Spectroscopy was also performed at a specific piezoelectric region to measure domain properties, such as the electric field required to flip the polarization direction (coercive voltage).
关键词： topography,atomic force microscopy,multilayered ceramic capacitor,electromechanics,piezoelectric microscopy,polarization,failure analysis
更新于2025-09-23 15:23:52
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A robust and versatile platform for image scanning microscopy enabling super-resolution FLIM
摘要： Image scanning microscopy (ISM) can improve the effective spatial resolution of confocal microscopy to its theoretical limit. However, current implementations are not robust or versatile, and are incompatible with fluorescence lifetime imaging (FLIM). We describe an implementation of ISM based on a single-photon detector array that enables super-resolution FLIM and improves multicolor, live-cell and in-depth imaging, thereby paving the way for a massive transition from confocal microscopy to ISM.
关键词： super-resolution,SPAD array,fluorescence lifetime imaging,confocal microscopy,image scanning microscopy
更新于2025-09-23 15:23:52