Computer-controlled double-beam scanning microspectrophotometry for rapid microscopic image reconstructions.

A method for automated collection of various specific data from an entire microscopical preparation and their quantitative evaluation is described. Its application to the study of neuronal connections is discussed in some detail. Brain sections are scanned using a computer-controlled microscope for reflectance, fluorescences or absorbance signals. Two illuminating beams are used, one of them being amplitude modulated. By means of a synchronous detection the two signals are recorded simultaneously: for example, in an autoradiograph, the reflectance (measuring the density of the silver grains in emulsion) and the absorbance (allowing to localize the underlying counterstained cells). The data are stored in a computer. Various off-line processing schemes allow the reconstruction of the data with respect to the corresponding spatial coordinates. Thus pseudo-three-dimensional, analogue or digital, graphic displays may be obtained in which the patterns of neuronal connections can be recognized and interpreted. A method for the detection of weakly labelled nerve fibres based on digital filtering is presented. The whole processing for a frontal section of the mouse brain (7 X 10 mm area) takes less than 1 h. In addition to the evaluation of microscopically labelled material (grains of autoradiographs, horseradish peroxidase, nucleic acids) the technique described has been successfully used for the study of naturally fluorescent intracellular components in living tissue cultures.     

Evaluation of characterization methods for thin sections of silver halide microcrystals by analytical electron microscopy

Silver halide microcrystals are usually composed of different phases with varying halide compositions. These microcrystals can be sectioned with an ultramicrotome with a certain thickness in a specific direction. The scanning electron microscopical analysis of these sections provides information on the internal halide profile.
During sample sectioning, many deformations can be induced in the material, and usually dominate the different scanning electron images.
By acquiring X-ray maps, it is possible to locate the different phases in the sections. This technique, however, is very time-consuming, and can only be used when the local halide variations are very high.
When an EDX line scan is acquired over the section, information on the position of the phases with different halide composition is obtained in a reasonable time, and shows no dependence on the section deformations. From this knowledge, several positions on the section are selected where local X-ray analyses are performed. After quantifying the results, the internal halide distribution is obtained. The analytical lateral resolution is about 30 nm for sections ≈100 nm thick.     

Quantitative method for estimating stain density in electron microscopy of conventionally prepared biological specimens

Stain density is an important parameter for optimising the quality of ultrastructural data obtained from several types of 3D electron microscopy techniques, including serial block-face electron microscopy (SBEM), and focused ion beam scanning electron microscopy (FIB-SEM). Here, we show how some straightforward measurements in the TEM can be used to determine the stain density based on a simple expression that we derive. Numbers of stain atoms per unit volume are determined from the measured ratio of the bright-field intensities from regions of the specimen that contain both pure embedding material and the embedded biological structures of interest. The determination only requires knowledge of the section thickness, which can either be estimated from the microtome setting, or from low-dose electron tomography, and the elastic scattering cross section for the heavy atoms used to stain the specimen. The method is tested on specimens of embedded blood platelets, brain tissue and liver tissue.     

Structural inspection and troubleshooting analysis of a lab-scale distillation column using gamma scanning technique in comparison with Monte Carlo simulations

In the present study, structural inspection and troubleshooting analysis of a lab-scale distillation column has been performed using gamma scanning technique and Monte Carlo simulations. MCNP4C Monte Carlo code has been used for simulations of the column and calculation of the computational density profile. The tested column is a one-pass tray type with 51 cm diameter. A Cs-137 sealed source and a 1 × 1 inch NaI (Tl) detector has been used for this gamma scanning process. According to the results, both experimental and simulation results showed the specification of trays and another section of the column accurately. Also, in addition to the flooding and damaged tray in the column, defects such as foaming with the density of 0.17 g/cm³ can be distinguished using this technique. Based on the results, using photopeak count approach the differences in the material attenuations can be better distinguished. The effectiveness of this approach in determination of malfunctions increases with the density of the material between the source and the detector. Analyzing the experimental and simulation results are indicative of the fact that the procedures and methods used in this work are quite suitable for improving the accuracy of the troubleshooting analysis based on gamma scanning technique.     

基于光学扫描测量数据的叶片型面偏差分析研究

为了满足叶片型面快速检测的需要,研究基于光学扫描测量数据的型面偏差分析技术.详细描述光学扫描测量的原理及数据采集过程,运用基于特征点的预配准和SVD-ICP算法来进行测量模型与CAD理论模型的配准定位,通过UG/API函数获取截面数据,对截面数据进行偏差计算,得到偏差情况统计结果.同时在偏差计算的过程中,针对叶片检测中截面各区域公差要求不一样的情况,提出一种基于轮廓线参数化的公差带设计方法,使得公差带的设计更好地符合实际检测的需要.基于上述算法,综合运用VC+ +,UG/API和VTK等工具,开发了叶片型面偏差分析软件,对于实现叶片型面的快速检测具有重要意义.     

端面约束单晶硅直梁MEMS扫描微镜应力特性研究

结合微机电系统(micro-electronic-mechanical system,MEMS)扫描微镜结构以及单晶硅材料正交各向异性的特点,理论分析端面约束单晶硅直梁MEMS扫描微镜的应力特性,得到器件工作时扭转梁上正确的应力分布及其特征,所得结论与有限元仿真结果进行比较.结果表明由于端部约束效应的影响,作用在MEMS扫描微镜上的驱动力矩分别通过纯扭转力矩和横向弯矩,在扭转梁上传递,导致在扭转梁产生正应力、附加剪切应力和扭转剪切应力,其中扭转剪切应力最大值位于扭转梁中部截面长边中点处,而正应力和附加剪切应力最大值分别位于约束端部截面四角处和截面短边中点处.随着扭转梁截面形状趋近于窄的矩形,端面约束效应会愈加明显,约束端部截面的正应力越大于其他两种应力,经过有限元分析获得与分析解较为一致的结果.     

A quantitative approach to cytoarchitectonics: Analysis of structural inhomogeneities in nervous tissue using an image analyser

Cytoarchitectonic investigations are based on the analysis of structural inhomogeneities in the neuronal composition of nervous tissue. Boundaries of brain regions are established at locations where local structural properties, such as density, shape, orientation or arrangement of the nerve cells, change. A quantitative approach requires a complete scanning of histological sections and the measurement of at least one of these properties using an automatic device. In contrast to recently developed stereological methods which result in unbiased estimates of parameters and which are based on efficient sampling procedures, measurements with automatic devices are biased due to section thickness and problems in the segmentation of nerve cell bodies. Automatic measurements are necessary, however, if a complete scanning of histological sections is required in order to localize areal or laminar boundaries. In this approach, the grey level index (GLI) is measured with a TV-based image analysing system from routine histological sections. Using Nissl-staining, this parameter is a biased estimate of the local volume density of Nissl-positive structures (cell bodies). The histological section is digitized into a GLI image by a scanning procedure. The GLI image is processed by image enhancement procedures in order to visualize the laminar pattern. Areal boundaries are localized at positions where this laminar pattern changes. GLI statistics of single brain regions can easily be evaluated by delineating these regions with a cursor in the GLI images of the sections. Information from a series of sections is compiled by specific application programs.     

快速造型技术中的反求工程

阐述反求工程在快速造型技术中所起的作用；目前反求工程在快速造型中常用的测量方法是三坐标测量仪法、激光三角形法、核磁共振（ＭＲＩ）和ＣＴ法、光栅法和自动断层扫描仪法；从测量原理、测量精度以及测量材料和范围等方面对测量方法作出分析和对比；总结在快速造型技术中所用的几种测量数据处理的方法，对比这些方法的原理、严谨及使用范围；提出适用于快速造型技术的反求工程系统所应具有的功能。     

Signal‐to‐noise ratio enhancement on SEM images using a cubic spline interpolation with Savitzky–Golay filters and weighted least squares error

A new technique based on cubic spline interpolation with Savitzky–Golay smoothing using weighted least squares error filter is enhanced for scanning electron microscope (SEM) images. A diversity of sample images is captured and the performance is found to be better when compared with the moving average and the standard median filters, with respect to eliminating noise. This technique can be implemented efficiently on real‐time SEM images, with all mandatory data for processing obtained from a single image. Noise in images, and particularly in SEM images, are undesirable. A new noise reduction technique, based on cubic spline interpolation with Savitzky–Golay and weighted least squares error method, is developed. We apply the combined technique to single image signal‐to‐noise ratio estimation and noise reduction for SEM imaging system. This autocorrelation‐based technique requires image details to be correlated over a few pixels, whereas the noise is assumed to be uncorrelated from pixel to pixel. The noise component is derived from the difference between the image autocorrelation at zero offset, and the estimation of the corresponding original autocorrelation. In the few test cases involving different images, the efficiency of the developed noise reduction filter is proved to be significantly better than those obtained from the other methods. Noise can be reduced efficiently with appropriate choice of scan rate from real‐time SEM images, without generating corruption or increasing scanning time.     

复合材料曲面构件缺陷超声三维成像方法

针对目前复合材料曲面结构缺陷检测技术存在的检测结果不直观、效率低等问题,提出一种基于超声相控阵的缺陷三维成像方法。使用三维激光扫描仪获取曲面的点云模型,通过平行截面法规划检测路径,然后使用相控阵轮式探头采集超声图像数据。利用均匀三次B样条函数拟合检测路径与曲面,根据扫查步长和图像序列关系计算超声图像数据点的空间位置以生成超声点云集。最后利用体素化降采样方法对超声检测结果进行重建,实现复合材料内部缺陷的三维成像。实验结果表明,本文方法的缺陷成像结果与CT检测结果的平均误差为1.14 mm,能够快速准确地重建缺陷的位置、形状与尺寸信息,实现复合材料曲面样件内部缺陷的精确表征。     

Distance measurement by circular scanning of the excitation beam in the two-photon microscope

We developed a method to measure relative distances with nanometer accuracy of fluorescent particles of different color in a two-photon scanning fluorescence microscope, with two-channel photon counting detection. The method can be used in the 10-500 nm range, for distances below the resolution limit of standard far field microscopy. The proposed technique is more efficient than the methods using raster scanning. To achieve maximum sensitivity in the radial direction, the excitation beam is moved periodically in a circular orbit with a radius of the size of the point spread function. The phase and the modulation of the periodic fluorescence signal, calculated by fast Fourier transform, gives the phase and the radial distance of the particle from the center of scanning. The coordinates of particles are recovered simultaneously in the two channels and the relative distance is calculated in real time. Particles can be tracked by moving the center of scanning to the recovered position, while measuring the distance from the second particle. Intensity data are saved and fitted later by a model accounting for light leakage between the channels. The total number of detected photons limited the accuracy of the position and distance measurement. Experiments demonstrating the advantages of the method were performed on fluorescent spheres and single dye molecules immobilized on quartz surface.     

State-of-the-art of non-destructive measurement of sub-surface material properties and damages

The possibilities of common non-destructive measuring techniques are reviewed in this paper for their applications in precision engineering. The grazing X-ray technique is believed to be a powerful improvement of the conventional X-ray techniques under both the diffraction mode and the fluorescent mode. Information of the crystallographic structure and chemical composition can be obtained to a nanometre resolution. Ultrasound can be used in scanning acoustic microscopy to give information on the physical or even chemical nature of superficial layers. Raman spectroscopy has now become an important tool for studying superficial structures, chemical composition and stresses in cyrstalline and amorphous materials: it is recommended to use this method especially for the investigation of monocrystalline silicon and germanium. The instrumented microindentation technique is a quasi- non-destructive technique for evaluating mechanical material properties like hardness and Young's modulus in a nanometre range. It can be used on any material that does not require special surroundings like a vacuum. Photothermal microscopy has been developed recently for the non-destructive testing of the local thermal properties of materials. By using the Mirage effect and its local measurement above the surface, a non-destructive depth profiling of surface damages can be obtained.     

组合梁有限元分析

通过ANSYS有限元的分析方法，针对相同几何截面下的单梁和不同材料组合梁，分析了在承受相同荷载作用下的变形及工作应力。ANSYS有限元分析过程比电测法更为方便、快捷、；隹确，是工程上解决应力、应变问题的另一种途径。     

NanoSIMS imaging of Bacillus spores sectioned by focused ion beam

Preparation and sectioning of bacterial spores by focused ion beam and subsequent high resolution secondary ion mass spectrometry analytical imaging is demonstrated. Scanning transmission electron microscopy mode imaging in a scanning electron microscope is used to show that the internal structure of the bacterial spore can be preserved during focused ion beam sectioning and can be imaged without contrast staining. Ion images of the sections show that the internal elemental distributions of the sectioned spores are preserved. A rapid focused ion beam top‐sectioning method is demonstrated to yield comparable ion images without the need for sample trenching and section lift‐out. The lift‐out and thinning method enable correlated transmission electron microscopy and high resolution secondary ion mass spectrometry analyses. The top‐cutting method is preferable if only secondary ion mass spectrometry analyses are performed because this method is faster and yields more sample material for analysis; depth of useful sample material is ～300 nm for top‐cut sections versus ～100 nm for electron‐transparent sections.     

层去图像法反求工程中的数据处理

提出层去图像法反求工程中的数据处理方法。首先采用多帧平均、中值滤波和保持细节平滑滤波对图像进行预处理,使图像质量得到恢复与改善,然后再进行图像分割和边缘检测,最后进行轮廓追踪有其多边形逼近。实践表明,所提出的数据处理方法具有较好的应用效果。     

Semi-automatic analysis of microscopic images of the human cerebral cortex using the grey level index

A new semi-automatic method is introduced which makes possible the quantitative examination of cresyl-fast-violet stained sections of the cerebral cortex. These sections are divided into measuring fields of 20 × 20 μm in a well-defined area. A scanning procedure then automatically determines the grey level index of every single field. The image of the histological section can be compared with the grey level index data matrix by means of a plotted image, the density of the plotted points of which is proportional to the measured grey level index. By overlaying this plotted image with the original section, the boundaries of the laminae can be fixed in the data matrix. The grey level index profiles—the grey level data plot as a function of the cortical depth—after smoothing and standardization are typical of special cortical areas. The mean thickness and the mean portion of the grey level index profile of the different laminae of the striate area are compared with data from other publications.     

Investigation of fluorescent lamp phosphors using the combined CL and EDS modes of the SEM

The phenomenon of cathodoluminescence (CL) potentially offers the ideal tool for studying the phosphor materials used in fluorescent lamps, since it can be used directly on processed or unprocessed powders, on coatings in tubes, or on sections cut from tubes. Using examples of both single component materials and multi-component blends, it is demonstrated how a relatively unsophisticated dispersive CL system attached to a scanning electron microscope (SEM) can be used expediently in the extensive study of such phosphors. These studies can be significantly enhanced when other complementary modes of the SEM (e. g. the energy-dispersive x-ray analysis facility) are combined with the CL mode. The strength of the combined technique lies in the major role it can play in materials and processing aspects of the powders themselves, in the processing of the lamps (e.g. by optimising such parameters as coating thickness, packing density etc.), and in diagnostic studies of poor materials or lamps (e.g. by locating rogue particles/components and identifying their origin). The technique also provides a convenient method of studying the temperature stability of selected phosphors.     

Finite element simulation of flexible roll forming with supplemented material data and the experimental verification

Flexible roll forming is a promising manufacturing method for the production of variable cross section products. Considering the large plastic strain in this forming process which is much larger than that of uniform deformation phase of uniaxial tensile test, the widely adopted method of simulating the forming processes with non-supplemented material data from uniaxial tensile test will certainly lead to large error. To reduce this error, the material data is supplemented based on three constitutive models. Then a finite element model of a six passes flexible roll forming process is established based on the supplemented material data and the original material data from the uniaxial tensile test. The flexible roll forming experiment of a B pillar reinforcing plate is carried out to verify the proposed method. Final cross section shapes of the experimental and the simulated results are compared. It is shown that the simulation calculated with supplemented material data based on Swift model agrees well with the experimental results, while the simulation based on original material data could not predict the actual deformation accurately. The results indicate that this material supplement method is reliable and indispensible, and the simulation model can well reflect the real metal forming process. Detailed analysis of the distribution and history of plastic strain at different positions are performed. A new material data supplement method is proposed to tackle the problem which is ignored in other roll forming simulations, and thus the forming process simulation accuracy can be greatly improved.     

A protocol for 3D image reconstruction from a single image of an oblique section.

Oblique section 3D reconstruction can produce a 3D image of a sectioned crystal from a single electron micrograph. We describe here in detail a reconstruction protocol applicable to an electron micrograph of an oblique section through a 3D crystal. The protocol is described in six steps: (1) selection criteria for images, (2) preprocessing steps to correct for image defects, (3) determination of unit cell coordinates, (4) interpolation of strip images with correction for image distortions and crystal disorder, (5) production of a crystallographic serial section reconstruction, (6) correction for skewed sampling to produce an oblique section reconstruction. In addition, we explore Wiener filter deconvolution of the section thickness. We describe a method for determining the section thickness by comparing data from projections of the oblique section reconstruction with corresponding data from a thick longitudinal section. Several schemes for Wiener filter deconvolution are described that differ in the way information on the signal-to-noise ratio is used in the filter.     

Quantitative analysis of backscattered-electron contrast in scanning electron microscopy

Backscattered-electron scanning electron microscopy (BSE-SEM) imaging is a valuable technique for materials characterisation because it provides information about the homogeneity of the material in the analysed specimen and is therefore an important technique in modern electron microscopy. However, the information contained in BSE-SEM images is up to now rarely quantitatively evaluated. The main challenge of quantitative BSE-SEM imaging is to relate the measured BSE intensity to the backscattering coefficient η and the (average) atomic number Z to derive chemical information from the BSE-SEM image. We propose a quantitative BSE-SEM method, which is based on the comparison of Monte–Carlo (MC) simulated and measured BSE intensities acquired from wedge-shaped electron-transparent specimens with known thickness profile. The new method also includes measures to improve and validate the agreement of the MC simulations with experimental data. Two different challenging samples (ZnS/Zn(O_xS_1–x)/ZnO/Si-multilayer and PTB7/PC₇₁BM-multilayer systems) are quantitatively analysed, which demonstrates the validity of the proposed method and emphasises the importance of realistic MC simulations for quantitative BSE-SEM analysis. Moreover, MC simulations can be used to optimise the imaging parameters (electron energy, detection-angle range) in advance to avoid tedious experimental trial and error optimisation. Under optimised imaging conditions pre-determined by MC simulations, the BSE-SEM technique is capable of distinguishing materials with small composition differences.