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Optical microscopy is the gold standard technique used to confirm the diagnosis of scabies. Multiple diagnostic features of the pathogen Sarcoptes scabiei var. hominis (S. scabiei) can be identified under a microscope and classified into 3 categories: mites, eggs and fecal pellets. However, mite and eggshell fragments can also be observed, which have been ignored in the 2020 International Alliance for the Control of Scabies (IACS) Criteria and by most researchers. In this study, we propose a novel morphological classification method that classifies multiple diagnostic features into 5 categories and 7 subcategories. Our results revealed that 65.2% (1893 of 2896) of the positive cases were confirmed through the identification of mites, eggs or fecal pellets, whereas up to 34.6% (1003 of 2896) of the positive cases were confirmed through the identification of mite or eggshell fragments. Therefore, the important diagnostic values of mite and eggshell fragments should be emphasized. Importantly, for the first time, mite and eggshell fragments were classified into 7 subcategories, some of which are easily ignored or confused with contaminating artefacts. We believe that this novel morphological classification method will be beneficial for operator training in interpreting slides and in improving the 2020 IACS Criteria.
We present a fast and precise deep-learning architecture, which we term O-Net, for obtaining super-resolved images from conventional phase-modulated optical microscopical techniques, such as phase-contrast microscopy and differential interference contrast microscopy. O-Net represents a novel deep convolutional neural network that can be trained on both simulated and experimental data, the latter of which is being demonstrated in the present context. The present study demonstrates the ability of the proposed method to achieve super-resolved images even under poor signal-to-noise ratios and does not require prior information on the point spread function or optical character of the system. Moreover, unlike previous state-of-the-art deep neural networks (such as U-Nets), the O-Net architecture seemingly demonstrates an immunity to network hallucination, a commonly cited issue caused by network overfitting when U-Nets are employed. Models derived from the proposed O-Net architecture are validated through empirical comparison with a similar sample imaged via scanning electron microscopy (SEM) and are found to generate ultra-resolved images which came close to that of the actual SEM micrograph.
Three-dimensional fluorescence microscopy is a key technology for inspecting biological samples, ranging from single cells to entire organisms. We recently proposed a novel approach called spatially modulated Selective Volume Illumination Microscopy (smSVIM) to suppress illumination artifacts and to reduce the required number of measurements using an LED source. Here, we discuss a new strategy based on smSVIM for imaging large transparent specimens or voluminous chemically cleared tissues. The strategy permits steady mounting of the sample, achieving uniform resolution over a large field of view thanks to the synchronized motion of the illumination lens and the camera rolling shutter. Aided by a tailored deconvolution method for image reconstruction, we demonstrate significant improvement of the resolution at different magnification using samples of varying sizes and spatial features.
In this work, the technique and the pictorial materials employed by Claude Monet in Pink Water Lilies, presently housed at the National Gallery of Modern and Contemporary Art in Rome, were investigated. The painting underwent noninvasive investigations such as energy-dispersive X-ray fluorescence and visible reflectance spectroscopies. The combined use of these techniques allowed us to identify most of the inorganic pigments such as cobalt blue and violet, zinc oxide, cadmium yellow, vermilion, and mixtures. Particularly, the spectrophotometric curves allow for the detection of the anhydrous and hydrated chromium greens. Two micro-fragments of the painting were also examined with micro-Fourier transform infrared spectroscopy and the cross-sections obtained were analyzed with the optical microscope and with scanning electron microscopy (SEM) and energy-dispersive X-ray spectrometry (EDS). Fourier Transform Infrared spectroscopy analyses allowed us to recognize the animal glue used for priming the canvas, which was covered with a ground layer consisting of calcite and lead white mixed with an oil binder. A lipidic binder was also detected in the color layer. Optical microscopy and SEM-EDS were useful to retrieve information about the stratigraphy, the distribution of pigments, and a more complete palette identification of phosphate, arsenate, and magnesium arsenate cobalt violets, and the red lake was possible.
The CytoViva Enhanced Darkfield (EDF) Illuminator enables direct optical observation of nano-scale entities in a wide range of transparent and translucent sample environments, without requiring the use of labels or other markers. The improved performance is a result of a uniquely designed light path, which focuses maximum photon density on the sample, thus enabling superior signal-to-noise imaging performance. This high-performance capability not only provides users with previously unobtainable images, but it also allows established techniques such as hyperspectral and Raman microscopy to be used in new and exciting ways.
Three clayey materials named MY3, KK and KG originating from the Foumban region (west Cameroon) were analysed to determine their granulometry, plasticity, major-element chemistry and mineralogy. Dilatometric and ceramic behaviour were also investigated. Clays were shaped by uniaxial pressing in a steel mould. Shaped samples were heated at 1300, 1400 and 1500°C. The end products were characterized in terms of their density, porosity and compressive strength. Raw materials differ in terms of their mineralogical composition, grain-size distribution, Al2O3 content and the nature and abundance of impurities inducing specific thermal behaviour during dilatometric analysis and sintering tests. The final material properties may be related to the main features of the raw materials used.
Multispectral imaging of ore minerals under the microscope is a logical extension of quantitative colour analysis and microspectrophotometric analysis of minerals. This paper describes, step by step, how the proper calibration of a scientific video camera can be performed in order to obtain precise reflectance measurements at each pixel within the field of view. After having reviewed the different sources of noise and aberration, practical formulae are presented that allow for the acquisition of a set of images at different wavelengths in the visible spectrum.
The advantage of using a multispectral image acquisition system based on narrow bandwidth (10 nm) interference filters is discussed and quantitatively compared to colour imaging using tristimulus (red, green, blue) filters.
Images taken from major sulphide parageneses are shown as examples of well contrasted multispectral images. Finally, the potential for automatic identification of ore minerals is discussed with reference to supervised multivariate image classification algorithms similar to those used in remote sensing. Additional comments on extending the principles for handling optical anisotropy and developing a multiradial imaging system are made.
In this paper, the change detection of a fast turning specimen is studied at micro-level, whereas the images are acquired without stopping the rotation. In the beginning of the experiment, the imaging system is focused on the surface of the specimen. By starting the rotation of the specimen, the diameter of the specimen changes due to wear, which results in de-focusing of the imaging system. So the amount of blur in the images can be used as evidence of the wear phenomenon. Due to the properties of the microscope, the corners of the frames were dark and had to be cropped. So, each micrograph reflects only a small area of the surface. Nevertheless, techniques like stitching of multiple images can provide a significant surface area for micro-level investigation which increases the effectiveness of analyzing the material modification. Based on the results computer vision could detect a change of about 1.2 µm in the diameter of the specimen. More important is that we could follow the same locations of the surface in the microscopic images despite blurring, uneven illumination, change on the surface, and relatively a high-speed rotation.
This study introduces a passive autofocus method based on image analysis calculating the Bayes spectral entropy (BSE). The method is applied to optical microscopy and together with the specific construction of the opto-mechanical unit, it allows the analysis of large samples with complicated surfaces without subsampling. This paper will provide a short overview of the relevant theory of calculating the normalized discrete cosine transform when analyzing obtained images, in order to find the BSE measure. Furthermore, it will be shown that the BSE measure is a strong indicator, helping to determine the focal position of the optical microscope. To demonstrate the strength and robustness of the microscope system, tests have been performed using a 1951 USAF test pattern resolution chart determining the in focus position of the microscope. Finally, this method and the optical microscope system is applied to analyze an optical grating (100 lines/mm) demonstrating the detection of the focal position. The paper concludes with an outlook of potential applications of the presented system within quality control and surface analysis.
Compounds of poly(3-hydroxybutyrate) (PHB) and carbon black (CB) with CB content ranging between 0.5 and 10% were prepared in an internal mixer. The effect of heating and cooling rates on the crystallization and melting of PHB/CB compounds was investigated by differential scanning calorimetry (DSC), and its morphology analyzed by optical microscopy (OM). Results showed that PHB and its compounds partially crystallize from the melt during cooling and partially cold crystallize on reheating, with the amount of polymer crystallizing in each stage depending strongly on the cooling rate. Melting is usually shown in DSC scans as complex (double) peaks, which are influenced by the heating/reheating thermal cycles. The melting and cold crystallization temperatures, and the rates of phase change depend strongly on the cooling and heating rates and CB content. CB acts as a nucleating agent, promoting the melt and cold crystallization of PHB as well as increasing the number of spherulites, with a mild effect on the melting transition. Light microscopy images suggest that a secondary crystallization of PHB also occurs during storage at room temperature.
A resolution metric intended for resolution analysis of arbitrary spatially calibrated images is presented. By fitting a simple sigmoidal function to pixel intensities across slices of an image taken perpendicular to light–dark edges, the mean distance over which the light–dark transition occurs can be determined. A fixed multiple of this characteristic distance is then reported as the image resolution. The prefactor is determined by analysis of scanning transmission electron microscope high-angle annular dark field images of Si<110>. This metric has been applied to optical, scanning electron microscope, and helium ion microscope images. This method provides quantitative feedback about image resolution, independent of the tool on which the data were collected. In addition, our analysis provides a nonarbitrary and self-consistent framework that any end user can utilize to evaluate the resolution of multiple microscopes from any vendor using the same metric.
We apply common image enhancement principles and sub-pixel sample positioning to achieve a significant enhancement in the spatial resolution of a vertical scanning interferometer. We illustrate the potential of this new method using a standard atomic force microscope calibration grid and other materials having motifs of known lateral and vertical dimensions. This approach combines the high vertical resolution of vertical scanning interferometry and its native advantages (large field of view, rapid and nondestructive data acquisition) with important increases in lateral resolution. This combination offers the means to address a common challenge in microscopy: the integration of properties and processes that depend on, and vary as a function of observational length.
Studies on cultural metal artifacts can benefit greatly from microscopy techniques. The examination of microstructural features can provide relevant information about ancient manufacturing techniques, as well as about corrosion/degradation processes. In the present work, advantages of the use of multifocus imaging techniques in optical microscopy for the study of archaeological metals are presented. An archaeometallurgical study of a large collection of bronzes demonstrates the possibility of a microstructural study with no need for sample removal, which is a great advantage in the study of cultural objects. In addition, the study of mounted samples illustrates the advantages of the multifocus technique in the examination of particular corrosion features, with the possibility of three-dimensional reconstructions.
Selective hydrogenation is an important process in petrochemistry to purify feedstock for polymer synthesis. For this process, catalysts containing metallic palladium deposited with an eggshell distribution on porous alumina are usually employed. For this kind of catalyst, the activity is known to be in close relation with the thickness of the palladium crust. As palladium oxide is brown and alumina is white, the palladium distribution in a catalyst bead before the reduction step can be characterized by optical microscopy. We propose an original and automatic procedure of optical image analysis to obtain a fast and robust method to measure the mean crust thickness of a catalyst batch and the corresponding standard deviation. The approach is validated by two different methods. First, we compared the crust thickness with those obtained by electron probe microanalysis. Then, catalytic tests of four samples with varying palladium crust thicknesses were performed and confirmed the expected correlation between activity and crust thickness measured by optical microscopy coupled with image analysis.
Because of its great potential to provide data on contacts and overseas trade, ivory has aroused a great deal of interest since the very start of research into Iberian late prehistory. Research recently undertaken by the German Archaeological Institute in Madrid in collaboration with a number of other institutions has provided valuable contributions to the study of ivory in the Iberian Copper Age and Early Bronze Age. One of the archaeological sites that is contributing the most data for analysing ivory from the Copper Age in southern Iberia is Valencina de la Concepción (Seville), which is currently the focus of several debates on the development of social complexity. This article contributes to this line of research by providing new, unpublished evidence and by examining the significance of ivory craftsmanship in commercial, social, and ideological terms. It also assesses in greater detail the prominent part played by luxury ivory items as an expression of social status and power.
A gray cast iron specimen was investigated by color and chemical etching with optical and atomic force microscopy, and the effect of grain orientation on the effectiveness of etching was examined. It was proven that the grain orientation dependence of chemical and color etching is just the opposite, and that the specimen surface after color etching is not uniformly smooth. Explanation for the layer structure of the color etched iron specimen is given.
The microstructure of Ti-35Nb-7.2Zr-5.7Ta (TNZT) and Ti-35Nb-7.2Zr-5.7Ta-0.5B (TNZTB) alloys under different heat treatment conditions has been analyzed. The solution-treated and water-quenched TNZT sample consists mainly of β phase with a very small amount of fine athermal ω precipitate. Precipitation of α can be observed when solution-treated samples are directly aged at 580°C for 8 h. The microstructure of the samples subjected to single-stage aging at 300°C or 400°C consists of ω precipitates in equiaxed β grains. Second stage aging at 580°C for 8 h after first stage of aging at 300°C or 400°C results in the replacement of ω precipitates by secondary α. In all of these samples, the amount of ω or α phase was very small, and therefore they could not be detected by X-ray diffraction studies. However, analysis of selected area diffraction patterns obtained from transmission electron microscopy studies confirms their presence. The addition of boron leads to the formation of dispersed precipitates of TiB in the β matrix of the TNZT alloy and also refines the β grains in the microstructure. However, other microstructural features of the TNZTB alloy are similar to those of the TNZT alloy.
Complementary state-of-the-art optical, scanning electron, and X-ray microscopy techniques have been used to study the morphology of Apollo 11 lunar soil particles (10084-47). The combination of innovative lighting geometries with image processing of a through focal series of images has allowed us to obtain a unique collection of high-resolution light micrographs of these fascinating particles. Scanning electron microscopy (SEM) stereo-pair imaging has been exploited to illustrate some of the unique morphological properties of lunar regolith. In addition, for the first time, X-ray micrographs with submicron resolution have been taken of individual particles using X-ray ultramicroscopy (XuM). This SEM-based technique lends itself readily to the imaging of pores, cracks, and inclusions and allows the internal structure of an entire particle to be viewed. Rotational SEM and XuM movies have also been constructed from a series of images collected at sequential angles through 360°. These offer a new and insightful view of these complex particles providing size, shape, and spatial information on many of their internal features.
An oil painting by Claude Monet, Port-Goulphar, Belle-Ile
1887 (collection of the Art Gallery of New South Wales), was examined
to determine both the identity of the pigments used by the artist in
this painting and his technique of mixing colors and laying paint on
the canvas. The extremely complex construction of the painting was
revealed by optical microscopy, scanning electron microscopy (SEM),
energy dispersive X-ray analysis (EDS), and X-ray mapping (XRM)
analysis of cross sections of paint flakes excised from damaged regions
of Port-Goulphar, Belle-Ile. Nine different pigments were
found on the painting. Many of the identified colors were modern
pigments that became available only late in the 19th century as a
result of scientific advances in pigment chemistry. Although similar
colors were available in a natural mineral form, they lacked the vivid
color of their manufactured counterparts. The use of these new
synthetic metallic oxide colors by Monet accounts for the brilliance of
his paintings. In addition, a separation between successive paint
layers was observed in some areas of paint chip cross sections,
indicating that oil-based paint was applied to paint that had dried,
and consequently, Port-Goulphar, Belle-Ile was painted over a
long period of time. This observation is contrary to the general
perception of Monet's technique of painting freely and quickly.
We probe the mesophase transitions and layer structures in thin ordered smectic liquid crystalline elastomer films by means of x-ray diffraction and optical microscopy. Oriented elastomer films of submicrometer thickness are produced by crosslinking freely suspended smectic polymer films. After crosslinking, the mesomorphism is similar to that of the precursor polymer. Smectic layers align parallel to the film plane. The layer spacing increases with temperature in the SmC* phase while it decays above the SmC*-SmA transition.