Image analyser measurements of coal reflectance

The application of an image analyser to the rapid measurement of coal reflectance histograms is described. The ability of an image analyser to utilize a two-dimensional image allows the coal only to be represented in the reflectance histogram, with contribution from the resin used in particulate block specimens eliminated. While the accuracy of reflectance measurement with an image analyser is not as great as can be obtained with a spot photometer system, sufficient accuracy is obtainable for rank determination.     

Analysis of coal blends by automated microscopy

The relative efficiency of coal blend analyses by automated reflectance measurement techniques is compared with an optimum point counting method. The influence of overlapping reflectance distributions and the advantage of image analyser systems over point reflectance measurements are evaluated. Automated point reflectance measurement techniques are feasible for coal blend analysis provided the measurement time is sufficiently small, while image analyser instruments can gain an advantage by reducing the particle top size in the specimen preparation.     

Characterization of coals by automated optical image analysis 1. Vitrinite reflectance

An increasing awareness of the importance of petrographic characterization of coals to efficient selection of coals for pulverized coal combustion and metallurgical coking, has highlighted the need to improve slow and subjective optical microscopic procedures. Automated image-analysis procedures to measure vitrinite random reflectance are examined here in some detail, giving particular attention to inertinite-rich coals. It is shown by consideration of intra- and inter-particle reflectance variance that a given accuracy for vitrinite mean random reflectance (R_v) can be achieved by selection of an appropriate surface sampling procedure. The repeatability of R_v for single coals is similar to that for manual microscopy, but the reproducibility, as established by an international interchange exercise, is not yet good enough to specify a standard procedure. In some coals even vitrinite sub-macerals can be distinguished; however, caution is required when extending this method to vitrinite reflectance distributions of blends containing different rank coals or of heat-altered coals.     

Apparent versus true density for the volume-to-weight transformation in coal blends

This work reviews the procedures involved in the conversion of the petrographic characterization data of blended coals into weight percentages of the single coals in the blends, the selection of the most suitable parameter for describing the density of the coals being critical. First, the fundamentals of the volume-to-weight transformation of the petrographic data were developed by means of mathematical formulae. This part of the procedure highlighted the need to use a density value adapted to the resolving power of the optical microscope. It was found that mercury penetration at 5 MPa, as determined by mercury porosimetry, was best suited for this purpose. A literature survey of published coal density data revealed that the use of correlations between coal density and rank was not an accurate enough procedure to deal with the relatively low corrections involved in the transformation of petrographic data. In the end, three binary blends were prepared from six carefully selected coals and polished particulate blocks of these blends were petrographically analysed to determine their compositions on a volume per cent basis. The best fit of these volume percentages with the weight percentages used in the formulation of the blends was found for a density correction based on mercury penetration at 5 MPa, in agreement with the empirical analysis.     

Reconstruction of the complex reflectance function in acoustic microscopy

We describe the theory and practical implementation of an iterative computer algorithm (an extension of the Gerchberg Saxton algorithm, originally developed for electron microscopy) for enabling an estimate of the complex reflectance function of a material to be reconstructed from measured values of only the magnitude of the response of an acoustic microscope (i.e. without the phase of the transducer voltage). Results are presented for eight materials measured with a spherical lens (at 320 MHz) and five with a cylindrical line-focus lens (at 210 and 228 MHz).     

Differentiation of live‐viable versus dead bacterial endospores by calibrated hyperspectral reflectance microscopy

This paper describes the use of hyperspectral imaging microscopy (HIM) for the characterization and differentiation of live viable versus dead/non‐viable bacterial endospores for two species of Bacillus. To accomplish this, endospore‐forming Bacillus were cultured and differentiated into endospores. Non‐viable endospores were produced using sporicidal methods representing standard decontamination procedures incorporating chlorine and peroxide. Finally, endospore samples were lyophilized to prepare them for spectral analysis. Prior to HIM, baseline spectral reflectance characterizing the endospores was measured using an ASD (400–900 nm) reflectance spectrometer. These data were used to calibrate the resulting spectral image data. HIM data comprising 32 images ranging from 400 to 720 nm (visible to near infrared) were recorded using a C‐mounted VariSpec hyperspectral camera attached to an epifluorescent microscope. The images produced by the system record the reflectance and absorption features of endospores based on the structure of the outer coat. Analysis of the HIM data was performed using accepted image and spectral processing routines. Where peroxide was the sporicide, changes in the outer endospore coat contributed to structurally significant visible and near infrared signature differences between live‐viable versus dead, non‐viable endospores. A statistical test for divergence, a method for scoring spectral structural diversity, also showed the difference between viable and non‐viable peroxide killed endospores to be statistically significant. These findings may lead to an improved optical procedure to rapidly identify viable and non‐viable endospores in situations of decontamination.     

An evaluation,by Fourier analysis,of some of the errors introduced in reflectance measurements

In order to evaluate the errors which can be introduced in the measurement of reflectance, we have made a study of data obtained for several minerals and in two immersion media on turning the microscope stage to a succession of positions relatively orientated by 45°. Fourier analysis of the results allows an estimate of the different errors as a function of the periodicity with which they appear on a full rotation of the stage. The mean magnitude of two of the Fourier coefficients is shown to provide a measure of the quality of the measurements further allowing evaluation of the equipment used, the orthogonality of the sample in relation to the incident light, and other errors of different periodicity.     

Geometrical characterization of fluorescently labelled surfaces from noisy 3D microscopy data

Modern fluorescence microscopy enables fast 3D imaging of biological and inert systems alike. In many studies, it is important to detect the surface of objects and quantitatively characterize its local geometry, including its mean curvature. We present a fully automated algorithm to determine the location and curvatures of an object from 3D fluorescence images, such as those obtained using confocal or light‐sheet microscopy. The algorithm aims at reconstructing surface labelled objects with spherical topology and mild deformations from the spherical geometry with high accuracy, rather than reconstructing arbitrarily deformed objects with lower fidelity. Using both synthetic data with known geometrical characteristics and experimental data of spherical objects, we characterize the algorithm's accuracy over the range of conditions and parameters typically encountered in 3D fluorescence imaging. We show that the algorithm can detect the location of the surface and obtain a map of local mean curvatures with relative errors typically below 2% and 20%, respectively, even in the presence of substantial levels of noise. Finally, we apply this algorithm to analyse the shape and curvature map of fluorescently labelled oil droplets embedded within multicellular aggregates and deformed by cellular forces.     

Correlative light and backscattered electron microscopy of bone--part II: automated image analysis

Detailed studies of biological phenomena often involve multiple microscopy and imaging modes and media. For bone biology, various forms of light and electron microscopy are used to study the microscopic structure of bone. Integrating information from the different sources is necessary to understand how different aspects of the bone structure interact. To accomplish this, methods were developed to prepare and image thin sections for correlative light microscopy (LM) and backscattered electron imaging in the scanning electron microscope (BSE-SEM). Images of the same fields of view may then be analyzed for degrees of relationships between specimen features not observed by LM or SEM alone. These methods are applied here to study possible associations between the degree of bone mineralization and pattern of collagen fiber orientation in the mid-shaft of the human femur. The "relational images" obtained allow us to examine the relationship between these two variables, both objectively and quantitatively.     

Ability of quality controllers to detect standard scratches on polished surfaces

The production of polished metal surfaces addresses a large number of fields such as medical prostheses, jewellery or moulds. These surfaces are checked by people who rarely agree on the definition of the right geometry limit of scratches. The aim of this work is to study human vision of scratches. The first step was to produce calibrated scratches barely visible by the human eye on polished metal surfaces in order to define standard samples for polishing quality control. A nanoindenter with a Berkovich tip was used to produce these scratches. These standard samples were shown to polishing controllers. They were asked to perform a research task leading to the establishment of two thresholds that characterize the visualization of scratches: a visibility threshold as well as an acceptability threshold.     

A comparison study of detecting gold nanorods in living cells with confocal reflectance microscopy and two‐photon fluorescence microscopy

Two‐photon fluorescence microscopy and confocal reflectance microscopy were compared to detect intracellular gold nanorods in rat basophilic leukaemia cells. The two‐photon photoluminescence images of gold nanorods were acquired by an 800 nm fs laser with the power of milliwatts. The advantages of the obtained two‐photon photoluminescence images are high spatial resolution and reduced background. However, a remarkable photothermal effect on cells was seen after 30 times continuous scanning of the femto‐second laser, potentially affecting the subcellular localization pattern of the nanorods. In the case of confocal reflectance microscopy the images of gold nanorods can be obtained with the power of light source as low as microwatts, thus avoiding the photothermal effect, but the resolution of such images is reduced. We have noted that confocal reflectance images of cellular gold nanorods achieved with 50 μW 800 nm fs have a relatively poor resolution, whereas the 50 μW 488 nm CW laser can acquire reasonably satisfactory 3D reflectance images with improved resolution because of its shorter wavelength. Therefore, confocal reflectance microscopy may also be a suitable means to image intracellular gold nanorods with the advantage of reduced photothermal effect.     

Representation of three-dimensionally reconstructed objects in electron microscopy by surfaces of equal density

A surface representation algorithm has been developed to visualize the results of three-dimensional reconstructions from projections in electron microscopy. It produces an easily recognizable image of the reconstructed particle and facilitates comprehension of the three-dimensional structure. Thus building of physical models can be avoided in many cases. If such a model is needed the algorithm can be used to establish the correct particle boundary in advance. Some further effort has been made to represent long fibres and visualize inner structures of objects, which are obscured by outer high-density regions. Shading may be used to enhance the depth perception from single views.     

Image division technique in pre-acquisition analysis of information content for automated microscopy

This article presents a method that allows for reliable automated image acquisition of specimens with high information content in light microscopy with emphasis on fluorescence microscopy applications. Automated microscopy typically relies on autofocusing used for the analysis of information content behaviour along the z-axis within each field of view. However, in the case of a field of view containing more objects that do not lie precisely in one z-plane, traditional autofocusing methods fail due to their principle of operation. We avoid this issue by reducing the original problem to a set of simple and performable tasks: we divide the field of view into a small number of tiles and process each of them individually. The obtained results enable discovering z-planes with rich information content that remain hidden during global analysis of the whole field of view. Our approach therefore outperforms other acquisition methods including the manual one. A large part of the contribution is oriented towards practical application.     

Evaluation and interpretation of grain density autoradiographs by reflectance photometry

The tritium content of nervous tissue sections is calculated from grain-density of autoradiographs without loss in anatomical information. The calculation is not based on determination of (expected) grain numbers but on photometric measurements (SKphot), interpreted as the result of the function of exposure time (T) and incorporated activity (β): SKphot = f (T, β). As experimental radiation standards homologous material (optic nerve preparations) with different activities determined by liquid scintillation counting has been used. Exposure times between 12 h and 28 weeks for these preparations with known radioactivities yielded a set of curves (nomogram) relating photometric values to exposure times. At any given exposure time the knowledge of photometric readings and related radioactivities within that set of curves enabled the development of a function covering all photometer readings within the range of the nomogram. Thus, any photometric reading could be correlated to the corresponding radioactivity. Such a function has been applied for the transformation of photometric values obtained from particular brain-section areas exposed for 16 weeks. Since the results are given in Bq, they may be related and, if necessary, corrected to results obtained by other tritium detection methods. Terminology used in the text: experimental radiation source = experimental standards prepared from optic nerve sections; standard radiation source = polymer reference source.     

基于RGB颜色信息聚类的光谱反射率重建

针对光谱反射率研究中因训练样本数据量大造成的冗杂等问题,提出了一种基于RGB信息进行聚类的样本分类方法。首先对颜色进行聚类并确定聚类个数,后对每一类光谱反射率使用BP神经网络分别进行重建。对于实验结果,使用ΔE00、均方根误差（RMSE）以及适应度系数等标准进行评价,同时与主成分分析算法进行对比。从实验分析可得出,在聚类数目为7时光谱反射率重建效果最好,这时的平均CIE2000的色差为0.836,平均RMSE为0.014 9,平均适应度系数为99.82%,并且,在最后对重建色差较大的色块进行了优化处理。实验证明,颜色聚类方法可以很好的应用于光谱反射率重建。     

Texture characteristics of unpolished and polished aggregate surfaces

In order to investigate evolution of polishing aggregate surfaces on an aggregate wear index (AWI) wear track specimen, experimental texture measurements and data dependent system (DDS) approach were utilized to model and analyze elevation profiles collected from unpolished and polished aggregate surfaces. It was found that the DDS approach was able to characterize the evolved macrotexture and microtexture. The polishing effect induced by the interaction between tire tread and aggregate surfaces was found to reduce the microtexture roughness significantly, but showed little influence on the macrotexture. This does not imply that the macrotexture plays little role in tire tread friction. It was also found that polishing effect presented a strong relationship with grain size existing on aggregate surfaces.     

An algorithm selection methodology for automated focusing in optical microscopy

Autofocus systems are essential in optical microscopy. These systems typically sweep the sample through the focal range and apply an algorithm to determine the contrast value of each image, where the highest value indicates the optimal focus position. As the optimal algorithm may vary according to the images' content, we evaluate the 15 most used algorithms in the field using 150 stacks of images from four different kinds of tissue. We use four measuring criteria and two types of analysis and propose a general methodology to apply to select the best fitting algorithm for any given application. In this paper, we present the results of this evaluation and a detailed discussion of different features: the threshold used for the algorithms, the criteria parameters, the analysis used, the bit depth of the images, their magnification, and the type of tissue, reaching the conclusion that some of these parameters are more relevant to the study than others, and the implementation of the proposed methodology can lead to a fast and reliable autofocus system capable of performing an analysis and selection of algorithms with no supervision required.     

Mapping the chemistry of resinite,funginite and associated vitrinite in coal with micro‐FTIR

Micro‐FTIR mapping is a powerful tool for nondestructive, in situ chemical characterization of coal macerals at high resolution. In this study, the chemistry of resinite, funginite and associated vitrinite is characterized via reflectance micro‐FTIR for Cenozoic high volatile C bituminous coals from Colombia. In comparison with the micro‐FTIR spectra of vitrinite and inertinite, the corresponding spectra of liptinite macerals in the same coals are characterized by stronger aliphatic CH_x absorbance at 3000–2800 and 1460–1450 cm^?1, but less intense aromatic C=C ring stretching vibration and aromatic CH_x out‐of‐plane deformation at 700–900 cm^?1. The aliphatic components in resinite have the longest carbon chains and are least branched, bestowing the highest hydrocarbon generation potential on resinite among the three macerals studied. In contrast, funginite exhibits the strongest aromatic character, the highest aromaticity, the lowest ‘A’ factor values and the lowest C=O/C=C ratios among the three maceral groups. Vitrinite generally displays intermediate chemical characteristics. Reflectance micro‐FTIR mapping of coal samples further confirms the aliphatic character of resinite and the aromatic nature of funginite. In addition, chemical mapping of resinite and adjacent vitrinite shows that vitrinite immediately adjacent to resinite displays higher aliphatic CH_x stretching intensity than more distant vitrinite, suggesting that chemical components from resinite can diffuse over short distances into adjacent vitrinite, specifically causing hydrogen enrichment. It needs to be pointed out, however, that the region of influence is localized and limited to a narrow zone, whose extent likely depends on resinite's properties, such as its size and aliphatic material content. This way, the chemical map of resinite and associated vitrinite provides direct evidence of the intermaceral effects occurring during the peat forming stage or during later coalification. No influence of funginite (primarily fungal spores and sclerotia) on the chemistry of adjacent vitrinite has been demonstrated, which is likely due to the highly aromatic structure of this type of funginite.     

红外镜反射法测定涂料薄膜的分子取向

国家纳米技术与工程研究院研发出的防污涂料是一种超低表面能氟碳涂料。使用红外镜反射光谱法可以快速判断该涂料的含氟基团在固体表面上呈定向分布。