双光源投影法细丝直径测量

提出一种细丝直径在线测量方法,利用半导体激光作为点光源,发出球面波;通过细丝后在其后菲涅耳衍射区留下阴影,测量阴影宽度可得细丝直径;同时,利用两个相距一定距离的点光源对同一细丝投影,测量两个投影之间的距离可得细丝位置的晃动量,从而校正直径测量值。     

基于双频彩色光栅投影测量不连续物体三维形貌

为减少多频条纹相移法进行三维形貌测量时的投影条纹数量,提高测量速度, 提出了采用双频四步相移彩色条纹投影技术来准确测量具有台阶状不连续或孤立物体表面三维形貌的方法。利用计算机将高低两种不同频率的正弦条纹分别输入彩色图像的红色和蓝色通道合成为彩色条纹,由数字视频投影仪将四步相移彩色条纹投影到被测物体,然后利用彩色CCD或CMOS相机采集4幅彩色条纹图并存储于计算机中。基于色彩分离技术得到8幅两种不同频率的四步相移条纹图并由图像灰度算法获得1幅反映背景的灰度图像;由低频条纹确定台阶状的物体边缘,高频条纹计算物体的形貌,并通过背景图像的二值化定位阴影和不连续区域。提出的方法仅用4幅彩色图像完成了台阶状不连续或孤立物体三维形貌的准确测量。与现有的多频4步相移条纹投影形貌测量技术需要8~12幅图像相比,该方法有效地减少了条纹投影与图像采集的数量。     

条纹投射系统深度标定中的数据处理算法分析

条纹投射测量系统深度标定的目的是建立条纹位相和被测物面深度之间的映射关系.标定过程中,不同的数据处理模型和算法对系统最终的测量精度有显著影响.分析比较了标定中几种常用的数据处理方法的特点,包括精确几何模型及其线性化模型,多项式逼近模型,在噪声条件下的误差分布特点,以及节点选择对多项式逼近模型标定精度的影响等.通过数值模拟和实验验证了分析结果的正确性.     

Diffraction effect of x-rays on the accuracy of quantitative analysis of crystals

In order to understand an apparent discrepancy of concentration of constituent elements in the quantitative analysis of thin KCl crystals with an energy dispersive X-ray analyzer, which has been partly explained by fluorescent X-ray excitation, more detailed experiments were carried out on binary alkali halides and tricomponent amorphous material by changing the azimuthal angle. Intensity ratios of X-rays from the constituent elements in KCl and KBr single crystals varied considerably according to the rotation of the specimens. The observed differences between maximum and minimum intensity ratios were 15% for a thin KCl crystal about 100 nm thick and 20% for a bulk KCl crystal, which corresponded to 4.1% and 4.5% differences in atomic concentration, respectively. It was ascertained that for amorphous materials such as Co₈₇Zr₅Nb₈, such a variation of the intensity ratio of X-rays was not observed. It is thus proved that the variations of the intensity ratios of X-rays from the constituent elements with azimuthal angle for KCl and KBr are attributed to the diffraction effect of characteristic X-rays generated in crystalline specimens. This effect, which is the same as that in the production of Kossel patterns, is one of the essential factors limiting the accuracy of the analysis.     

条纹投射测量标定中摄像机畸变图像的修正

在条纹投射测量系统的标定过程中对摄像机畸变图像进行修正,能有效地提高系统的标定精度。采用改进的Harris算子检测标定板上特征点的亚像素坐标,通过预标定获得特征点空间坐标,并应用BP神经网络技术校正畸变图像。仿真实验表明该方法能简单有效地提高了标定系统的精度。     

复杂背景成像条件下运动点目标的轨迹提取

为改进空间目标天基实时监视能力,研究了美国在空间中段实验卫星上搭载的对空间目标进行在轨检测与跟踪的信号处理器的工作方案.针对该处理器在目标运动轨迹提取阶段采用的先二元速度滤波再能量累积判决的“筛选-确认”解决方案,提出了两点改进方法:一是在“筛选”阶段增加运动速度约束条件,二是在“确认”阶段增加样本均差约束条件,使其在降低虚警概率的同时提高检测概率,从而普遍适用于复杂背景成像条件下运动点目标轨迹的提取.最后,利用实际获取的云层背景图像数据库仿真生成了包含多运动目标的时序图像序列,并以此作为输入信号源分析比较了原算法与改进算法的性能差异.仿真实验结果表明:改进算法在二元速度滤波后候选条痕减少到原算法的50％以下,处理器运行时钟周期数从8.0×106次降低到7.1×106次;最终检测结果显示,改进算法判决门限的合理取值范围增加到20左右,可以实现对多运动目标的实时检测.     

超声乳腺肿瘤图像中种子点的自动定位研究

利用种子区域增长对超声乳腺肿瘤图像进行分割是一种常用的计算机辅助诊断方法。为实现种子点的自动快速定位,满足实时在线分割图像的需求,根据超声乳腺肿瘤图像的结构特征,综合图像的灰度因素和空间因素,提出了一种基于迭代四叉树分解的算法。该算法将满足特定阈值的图像分裂转化为寻找种子区域,以实现种子点的自动定位。对105幅超声乳腺肿瘤图像进行了实验验证,结果表明,该算法准确率能够达到94.28%,平均耗时2.97 s,不但满足了种子点的自动定位于图像肿瘤内部,而且需要调整的参数少,其定位效率要高于人工选择。     

Use of Peldri II (a fluorocarbon solid at room temperature) as an alternative to critical point drying for biological tissues

A new chemical, Peldri II, is evaluated as a compound for drying soft biological tissues for scanning electron microscopy. Peldri II, a fluorocarbon, is a solid at room temperature and is a liquid above 25°C. Cells or tissues are embedded in Peldri II by immersing them in the liquid form and allowing it to solidify. Once solidified, Peldri II will sublime with or without vacuum to dry tissues, probably without introducing surface tension. Several types of cells and tissues have been examined to compare preservation with Peldri II and critical point drying techniques. No differences were detected between the two techniques when normal surface structures were examined. Peldri II appears to be a significant improvement over hexamethyldisilazane as a drying agent for scanning electron microscopy. It is also very convenient for drying large numbers of samples.     

Direct probing of solvent-induced charge degradation in polypropylene electret fibres via electrostatic force microscopy

Electrostatic force microscopy was used to directly probe solvent‐induced charge degradation in electret filter media. Electrostatic force gradient images of individual polypropylene electret fibres were used to quantify the extent of charge degradation caused by the immersion of the fibres into isopropanol. Electrostatic force gradient images were obtained by monitoring the shifts in phase and frequency between the oscillations of the biased atomic force microscopy (AFM) cantilever and those of the piezoelectric driver. Electrostatic force microscopy measurements were performed using non‐contact scans at a constant tip‐sample separation of 75 nm with varied bias voltages applied to the cantilever. Mathematical expressions, based on the capacitance of the tip‐sample system, were used to model the phase and frequency shifts as functions of the applied bias voltage to the tip and the offset voltage due to the fibre's charge. Quantitative agreement between the experimental data and the simplified model was observed.     

The effect of subsurface topography on secondary electron images from chromate pretreated aluminium surfaces

In secondary and scanning transmission electron microscopes, secondary electron images of surface films can be dominated by an image derived from electrons back-scattered from the interface between the film and the substrate. The extent of the domination has been established by studying the variation in image obtained using primary beams of different energy and by platinum coating to enhance surface secondary electron emission. Studies of thicker films also established that chemical or structural difference within a film also lead to imaging effects. In general, 5 keV electrons are the most effective in producing subsurface and structural or chemical imaging effects.     

Charge contrast imaging of material growth and defects in environmental scanning electron miscroscopy--linking electron emission and cathodoluminescence

An electron-based technique for the imaging of crystal defect distribution such as material growth histories in non- and poorly conductive materials has been identified in the variable pressure or environmental scanning electron microscope. Variations in lattice coherence at the meso-scale can be imaged in suitable materials. Termed charge contrast imaging (CCI), the technique provides images that correlate exactly with emitted light or cathodoluminescence in suitable materials. This correlation links cathodoluminescence and an electron emission. The specific operating conditions for observation of these images reflect a complex interaction between the electron beam, the positive ions generated by electron-gas interactions in the chamber, a biased detector, and the sample. The net result appears to be the suppression of all but very near surface electron emission from the sample, probably from of the order of a few nanometres. Consequently, CCI are also sensitive to very low levels of surface contaminants. Successful imaging of internal structures in a diverse range of materials indicate that the technique will become an important research tool.     

Bordetella adenylate cyclase toxin induces a cascade of morphological changes of sheep erythrocytes and localizes into clusters in erythrocyte membranes

Adenylate cyclase toxin (CyaA) of Bordetella pertussis penetrates the membrane of eukaryotic cells, producing high levels of intracellular cAMP, as well as hemolysis that results from the formation of cation-selective toxin channels in the membrane. Using several microscopical approaches we studied the effects of CyaA action on the morphology of sheep erythrocytes during early phases preceding lysis and examined localization of CyaA molecules within the erythrocyte membrane. CyaA induced a cascade of morphological changes of erythrocytes, such as shrinkage, formation of membrane projections, and blebs and swelling. The use of an enzymatically inactive CyaA-AC- toxoid that is unable to produce cAMP and of a CyaA-E581K mutant exhibiting higher hemolytic activity than with CyaA showed that the hemolytic activity is responsible for the induction of morphological changes of erythrocytes. Further, immunolabeling of inserted CyaA-232/FLAG molecules with specific anti-FLAG antibodies and IgG-gold particles indicated a clustered distribution of CyaA molecules in erythrocyte membrane. This was confirmed by immunofluorescence and confocal microscopy, which revealed uniform stoichiometry of CyaA clusters, suggesting CyaA binding into specific domains in erythrocyte membrane. Indeed, a decrease of CyaA binding after cholesterol depletion of erythrocytes suggests toxin targeting and binding to membrane microdomains (rafts).     

A method of direct particle deposition on a carbon planchet to study mineral dust in human lung by scanning electron microscopy

Following Na-hypochlorite digestion of lung tissue, mineral particles extracted in the chloroform layer were deposited directly on a pre-smoothed carbon planchet for combined scanning electron microscopy and X-ray energy dispersive spectrometry (SEM and XEDS). Total mineral particle counts were obtained, and detailed physical characteristics of the fibrous particles were documented at 600, 1,500, 4,500 and 9,000 x in three lungs without, and one lung with, histories of occupational exposure. This preparation method was simple, collected more than 99% of identifiable mineral particles in the chloroform layer, gave excellent object to background contrast without heavy metal coatings, and was suitable for XEDS. Comparable fibrous particles from the chloroform layer could also be studied by selected-area electron diffraction to complement the results of XEDS. By this method, we found particles or fibers larger than 0.1 μm were readily counted and measured at 4,500 x. At 600 x, ferruginous bodies were found to be more than twice in number than when sought for by light microscopy. It was determined that 4,500 x is the most efficient magnification to examine and diagnose this type of specimen. The present study illustrates the importance of determining the most efficient magnification to be utilized in particle counts.     

Mass loss rate in collodion is greatly reduced at liquid helium temperature

The mass thickness of collodion films has been monitored at several temperatures, under conditions typical of electron microscopy, through the use of an electron energy loss spectrometer. Compared to room temperature, only a five-fold reduction in the rate of mass loss was observed through the use of a commercial liquid nitrogen cooled stage; in contrast, the rate of mass loss was reduced more than one hundred fold when these films were held at liquid helium temperature.     

Explorative statistical analysis of planar point processes in microscopy

Basic methods of explorative statistical analysis for stationary and isotropic planar point processes are briefly and informally reviewed. At the explorative level, planar point patterns may be characterized in terms of the intensity, the K-function and the pair correlation function. These second-order functions enable one to classify a given point process as completely random, clustering or repulsive. The repulsive behaviour may be quantified by an estimate of the hard-core distance. In the exploratory approach, the statistics are essentially free from model assumptions. Second-order spatial functions have been estimated to characterize genuine planar point processes in the macroscopic domain, for example in forestry, geography and epidemiology. For light microscopy and transmission electron microscopy, two situations are distinguished, which may be summarized as the genuine planar case and the stereological case. In the genuine planar case, a direct interpretation of the results of spatial statistics is feasible. Here, monolayers in cell culture, intramembranous particles on freeze fracture specimens and amacrine cells of the retina are mentioned as examples. In the stereological case, point patterns are generated by sections through 3D structures. Here the observed point patterns may arise as the centres of sectional profiles of particles, or as centres of sectional profiles of spatial fibre processes. In both situations, exploratory spatial point process statistics allow a quantitative characterization of sectional images for the purposes of group comparisons and classification. Moreover, for spatial fibre processes it has recently been shown that the observed pair correlation function of the centres of the fibre profiles is an estimate of the reduced pair correlation function of the fibre process in 3D. Hence for fibre processes a stereological interpretation of point process statistics obtained from sections is an additional option.     

A simple method for the acquisition of high-quality digital images from analog scanning electron microscopes

A method is described for converting video signals of analog scanning electron microscopes (SEMs) into digital images of high quality. A plug-in card commercially available for personal computers is used for the on-line analog/digital conversion. A Windows application program written by the authors, together with low-level software drivers supplied with the plug-in card, allow digital images to be recorded, to be displayed simultaneously on the computer monitor and to be saved as a file in a standardized format. Compared to conventional photographic images obtained from the SEM camera system, the digital images possess superior sharpness of outline, excellent image definition, diminished noise and well-defined grey-scale tones. This method provides SEM images of high quality for less than $1000 from most older analog SEMs. In addition, the advantages of digital image processing can be applied to analog SEMs, including contrast enhancement, digital filtering and multichannel recording.     

Measuring the randomness of micro- and nanostructure spatial distributions: Effects of scanning electron microscope image processing and analysis

The quantitative characterisation of the degree of randomness and aggregation of surface micro- and nanostructures is critical to evaluate their effects on targeted functionalities. To this end, the methods of point pattern analysis (PPA), largely used in ecology and medical imaging, seem to provide a powerful toolset. However, the application of these techniques requires the extraction of the point pattern of nanostructures from their microscope images. In this work, we address the issue of the impact that Scanning Electron Microscope (SEM) image processing may have on the fundamental metric of PPA, that is, the Nearest Neighbour Index (NNI). Using typical SEM images of polymer micro- and nanostructures taken from secondary and backscattered electrons, we report the effects of the (a) noise filtering and (b) binarisation threshold on the value of NNI as well as the impact of the image finite size effects. Based on these results, we draw conclusions for the safe choice of SEM settings to provide accurate measurement of nanostructure randomness through NNI estimation.     

Automated segmentation of wood fibres in micro‐CT images of paper

A novel algorithm has been developed and validated to isolate individual papermaking fibres in micro‐computed tomographic images of paper handsheets as a first step to characterize the structure of the paper. The three‐step fibre segmentation algorithm segments the papermaking fibres by (i) tracking the hollow inside the fibres via a modified connected component methodology, (ii) extracting the fibre walls using a distance transform and (iii) labelling the fibres through collapsed sections by a final refinement step. Furthermore, postprocessing algorithms have been developed to calculate the length and coarseness of the segmented fibres. The fibre segmentation algorithm is the first ever reported method for the automated segmentation of the tortuous three‐dimensional morphology of papermaking fibres within microstructural images of paper handsheets. The method is not limited to papermaking fibres, but can be applied to any material consisting of tortuous and hollow fibres.