Low-voltage transmission electron microscopy reveals SV40 viral particles within secretory granules in pancreatic cells

Novel approach in low voltage transmission electron microscopy (TEM) has revealed the presence of SV40 viral like particles in the secretory zymogen granules and in spherical membrane-bound dense bodies of SV40 infected pancreatic cells. The presence of SV40 antigen in these cellular compartments was confirmed by immunocytochemistry of the VP1 antigen. Visualization of the viral particles was only possible by examining ultrathin tissue sections with low-voltage TEM that significantly enhances imaging contrast. Results indicate that following infection of the cell entry and trafficking of the viral particles are present in unique cellular compartments such as ER, dense bodies, and secretory granules.     

Freeze-fracturing of biological material with a new specimen table

The extraction of glycoproteins labelled with 3H or 14C precursors: fucose and glucosamine, has been compared during processing of rat liver for transmission electron microscopy. Using 14C labelled fucose, approximately 6% of labelled macromolecules were extracted during processing including 3% during glutaraldehyde fixation, 1% during post-fixation and 2% during dehydration. Greater extraction (8%) occurs with glucosamine as a precursor, which may be attributed to hydrolysis of the more labile sialic acid residues of the glycoprotein molecules. Tritium labelled glycoproteins were more susceptible to extraction than 14C-labelled glycoproteins. The extraction of 3H-labelled glycoproteins as measured by liquid scintillation counting may prove difficult to interpret owing to the quenching from processing solutions.     

Quantification of small, convex particles by TEM

It is shown how size distributions of arbitrarily oriented, convex, non-overlapping particles extracted from conventional transmission electron microscopy (TEM) images may be determined by a variation of the Schwartz-Saltykov method. In TEM, particles cut at the surfaces have diminished projections, which alter the observed size distribution. We represent this distribution as a vector and multiply it with the inverse of a matrix comprising thickness-dependent Scheil or Schwartz-Saltykov terms. The result is a corrected size distribution of the projections of uncut particles. It is shown how the real (3D) distribution may be estimated when particle shape is considered. Computer code to generate the matrix is given. A log-normal distribution of spheres and a real distribution of pill-box-shaped dispersoids in an Al-Mg-Si alloy are given as examples. The errors are discussed in detail.     

Application of nanoparticle tracking analysis platform for the measurement of soot-in-oil agglomerates from automotive engines

Nanoparticle Tracking Analysis (NTA) has been applied to characterising soot agglomerates of particles and compared with Transmission Electron Microscoscopy (TEM). Soot nanoparticles were extracted from used oil drawn from the sump of a light duty automotive diesel engine. The samples were prepared for analysis by diluting with heptane. Individual tracking of soot agglomerates allows for size distribution analysis. The size of soot was compared with length measurements of projected two-dimensional TEM images of agglomerates. Both the techniques show that soot-in-oil exists as agglomerates with average size of 120 nm. NTA is able to measure particles in polydisperse solutions and reports the size and volume distribution of soot-in-oil aggregates; it has the advantages of being fast and relatively low cost if compared with TEM.     

近壁面超声空化微射流对微细颗粒的破碎作用

针对微细粉体“团聚”导致粉磨极限的问题,在液相环境下利用超声的分散和空化冲击作用,开展了近壁面超声空化微射流对微细颗粒破碎作用的研究。通过理论计算空化微射流冲击微细颗粒破碎的有效作用范围,从空化泡溃灭速度的角度分析了超声频率、声压幅值、介质尺寸等主要参数对微射流强度的影响;结合物料质量浓度、介质尺寸、介质面积和功率等影响因素的微细颗粒超声空化破碎正交试验,并利用SEM观测粉体形貌,分析了颗粒中位粒径D50、10%体积累积粒径D10和比表面积（SSA）等分布特性。参数组合优化后获得了粒径小于800目的微细颗粒,破碎率高达79.35%;粒径大于10 000目的极细颗粒产率高达12.84%。从提高微细颗粒破碎率的角度,发现介质面积是主要影响因素,功率次之,其次为介质尺寸和物料质量浓度。试验结果与理论研究成果基本一致,表明优化超声空化微射流参数与增加介质壁面面积等方法可有效提高微细颗粒的破碎率。     

Third generation wavelet for the extraction of morphological features from micro and nano scalar surfaces

This paper investigates wavelet theory and techniques for the extraction of morphological structures from micro/nano scalar surfaces. It will significantly extend the authors’ previous accomplishments in the programme (EU: SMT CT98-2209). This previous work identified problems in the extraction of morphological structures that requires further study. Under a new EPSRC found project, the present work tries to overcome these problems and facilitates more practical and feasible methodologies for the identification and extraction of morphological structures on micro/nano scalar surface textures.In the current work, a complex wavelet model is introduced to solve the problems of nanometre roughness analysis of surface texture with linear features and curve-like features, in which small shifts of the input signal can cause large variations in the distribution of energy between wavelet coefficients at different scales. As a result, linear and curve-like features on a surface topography can be identified and extracted with refined accuracy. Case studies are conducted using automotive cylinders and femoral heads to demonstrate the application of using the new wavelet model in the assessment of surface topography.     

An electron microscopic analysis on the ultra structural abnormalities in sperm of the common carp Cyprinus carpio L. inhabiting a polluted lake, Umiam (Meghalaya, India)

The present communication reports the ultra structural abnormalities in sperm of a fish species Cyprinus carpio inhabiting a polluted lake, Umiam in North‐East India. Transmission electron microscopy (TEM) revealed absence of differentiation between head and midpiece (neck) of some sperm while scanning electron microscopy (SEM) showed some sperm tails with highly reduced length and some sperm with folded tail. Abnormal shape of some sperm head was also revealed by Scanning electron microscopy. Detachment of membrane from some parts of the sperm head and an outward expansion of the same was observed from Transmission electron micrographs of transverse section of sperm head. The well developed mitochondria surrounding the cytoplasmic channel in the sperm tail, as observed in control were found to be drastically disorganized in fish inhabiting the polluted lake. The study suggests that the fish C. carpio inhabiting the polluted lake Umiam is under severe stress as far as its male reproductive system is concerned. The study further suggests that Electron microscopic approach is extremely important in the assessment of adverse effects of environmental pollution on fish tissue. Microsc. Res. Tech. 2011. © 2011 Wiley Periodicals, Inc.     

基于冗余提升形态小波的非对称表面特征提取研究

针对提升小波提取非对称表面形貌特征时存在平移变动性和较差的方向选择性,将具有平移不变性和良好的方向选择性的冗余提升形态Haar小波应用到非对称表面的特征提取中。开展了该形态小波对具有显著特征的非对称平顶珩磨的缸套内表面进行特征提取的分析;同时结合仿真的方法,研究了该形态Haar小波对一个模拟的具有一定长度沟槽特征的空间表面进行特征提取的效果。研究结果表明,冗余提升形态Haar小波对非对称表面形貌进行形态特征提取时,在各尺度重构的信号沿特征边缘几乎没有变动且无畸变,可实现准确提取。     

Extraction of carbon 14-labeled compounds from plant tissue during processing for electron microscopy

Loss of ¹⁴C-labeled compounds from bean leaf tissue was monitored during all the stages of routine specimen preparation. No significant differences in extraction were associated with the use of acetone, ethanol, or dioxane as dehydration fluids. Fixation at low temperature increased the loss of label. Prolonged fixation in glutaraldehyde increased the loss, but fixation in osmium solutions for periods as long as 4 hr had no influence on extraction. Buffer rinses and dehydration fluids caused appreciable amounts of label to be extracted. The use of propylene oxide as transition fluid resulted in low extraction. Some embedding media caused the loss of small amounts of labeled compounds, but one of the media tested (LR-white) extracted significant amounts of label.     

曲波域铁谱磨粒图像特征提取方法研究

曲波变换具有多尺度分析能力,与小波变换相比可更好地表达图像的曲线特征.为有效描述铁谱磨粒的形貌特征,提出一种曲波域图像特征提取方法.利用曲波变换将磨粒图像进行分解,得到不同尺度的曲波系数;根据曲波系数统计分布特点,采用广义高斯分布模型对细尺度和精细尺度曲波系数分布进行建模;提取粗尺度曲波系数的均值、标准差、能量和熵等统计特征,以及细尺度和精细尺度曲波系数的广义高斯分布模型参数描述磨粒特征.将提取的特征用于发动机典型磨粒识别,识别成功率达到了88.9%,表明该方法所提特征能很好地表达铁谱磨粒的形貌特征.     

Enhancing microscopy images of minerals through morphological center operator‐based feature extraction

To well enhance the mineral image and image details obtained by microscopes, an effective mineral image enhancement algorithm through feature extraction using the morphological center operator is proposed in this work. First, mineral image feature extraction based on the morphological center operator is proposed and discussed. Second, the multiscale extension of the mineral image feature extraction is given by using the multiscale structuring elements. Third, the important mineral image features at multiscales of image are extracted and used to construct the final mineral features for mineral image enhancement. Finally, the original mineral image is well enhanced through importing the extracted final mineral image features into the original mineral image. Experimental results on different types of microscopy images of minerals verified the effective performance of the proposed algorithm for microscopy mineral image enhancement. Microsc. Res. Tech., 2013. © 2012 Wiley Periodicals, Inc.     

Electron microscopic observation of the sagittal structure of Drosophila mature sperm

Observation of sperm development and determination of their morphological characteristics are very important to the understanding of phylogenetic relationships and the study of sperm function during fertilization. Although ultrastructural studies of sperm development in the testes of the fruit fly Drosophila have been performed, there are few reports describing electron microscopic morphology of mature sperm, that is, those released from the testes to the seminal vesicles. Here, we present the first report of the sagittal organization of Drosophila sperm head and neck regions by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The head and tail structures of a mature sperm, for example, the acrosome, nucleus, and flagellum, were easy to distinguish by the morphological characteristics of the sperm surface by SEM. The morphological relationships between the surface and internal structures of mature sperm were confirmed by observing longitudinal sections with TEM. Our approach overcame the technical difficulties involved in sample preparation for electron microscopic observation of the Drosophila mature sperm head, and therefore, this study serves as an important foundation for future genetic dissection of sperm ultrastructure and function in male sterile mutants. Microsc. Res. Tech. 77:661–666, 2014. © 2014 Wiley Periodicals, Inc.     

Use of atomic force microscopy (AFM) for microfabric study of cohesive soils

Microfabric reflects the imprints of the geologic and stress history of the soil deposit, the depositional environment and weathering history. Many investigators have been concerned with the fundamental problem of how the engineering properties of clay depend on the microfabric, which can be defined as geometric arrangement of particles within the soil mass. It is believed that scanning electron microscopy (SEM) and transmission electron microscopy (TEM) are the only techniques that can reveal particle arrangements of clayey soils directly; however, current research introduces a novel and more advanced technique, atomic force microscopy, to evaluate the microfabric of cohesive materials. The atomic force microscopy has several advantages over SEM/TEM for characterizing cohesive particles at the sub‐micrometre range by providing 3D images and 2D images with Z‐information used in quantitative measurements of soil microfabric using SPIP software, and having the capability of obtaining images in all environments (ambient air, liquids and vacuums). This paper focuses on the use of atomic force microscopy technique to quantify the microfabric of clayey soils by developing the criteria for average and maximum values of angle of particle orientation within the soil mass using proposed empirical equations for intermediate and extreme microfabrics (dispersed, flocculated).     

基于形态学和像素灰度归类的背景重建算法

针对传统背景提取方法在前景运动对象密度较高,亦或前景出现过于频繁时,提取到的背景图像会掺杂很多前景成分的问题,提出了一种基于形态学和像素灰度归类的背景重建算法。该算法通过三帧差分法和形态学把每帧图像分为前景对象和背景区域;然后再用像素灰度归类法只对背景区域进行背景重建,以期在前景对象密度较高时正确重建背景。实验结果证明,新方法能够在车辆密度较高的情况下正确生成背景。     

A proposal for dichroic experiments in the electron microscope

Building upon the similarities between inelastic electron scattering and X-ray absorption we show that dichroism can be observed in electron energy loss spectrometry (EELS) in the transmission electron microscope (TEM). Natural or magnetic linear dichroism can be studied in electron scattering experiment with definite wave vector transfer in the interaction.The detection of circular dichroism in the TEM relies on interferometric EELS in a particular scattering geometry that allows extraction of the mixed dynamic form factor from energy loss spectra. Similarities between dichroic signals in energy loss near edge structures and X-ray absorption near edge structures are discussed, and a new experimental setup for dichroic measurements in the TEM is proposed.     

Alterations of lymphocyte membranes during HIV-1 infection via multiple and simultaneous entry strategies

Human immunodeficiency virus type 1 (HIV-1) must bind to and enter lymphocytes to replicate and cause the acquired immunodeficiency syndrome. The association of viral particles with the lymphocyte plasma membrane may vary according to a multitude of unknown variables, including lymphocyte membrane receptor mobilization, lipid raft aggregation, clathrin, caveolin, endosomes, microendosome-mediated penetration or penetration through a hole in the membrane. The time course of this delivery appears to be short. Fusion of the virion membrane and lymphocyte plasma membrane leads to destabilization of the lymphocyte membrane. Five morphological stages of membrane alteration were observed in the infected lymphocytes: (1) swelling, (2) splitting, (3) fusion, (4) breaking, and (5) thinning of the lipid bilayer. These plasma membrane alterations were not contributed by fixation artifacts, because the dimensions and distance between the subunits of the surface glycoprotein (SU, gp120) and the transmembrane glycoprotein (gp41) of the viral particles adjacent to the infected cells and processed at the same time remained unchanged. Destabilization of lipid raft patches in the lymphocyte plasma membrane by unknown variables may facilitate HIV-1 penetration of lymphocyte, and other cell types. This a combined review of the pertinent literature with our data showing that HIV-1 may take advantage of multiple penetration approaches simultaneously in the same cell type (H9) to overwhelm the infected cells. The ultrastructural details of H9 cultured cells infected in vitro with HIV-1 contribute to our understanding of viral particle association with the plasma membrane of infected cells.     

Use of the scanning electron microscope and Fourier methods in characterization of microfabric and texture of sediments

Behaviour of soils and sediments is generally assessed by laboratory tests of mass property relationships and semi-empirical equations. A basic approach calls for more detailed information concerning the properties of sediments and manageable methods of calculation. The scanning electron microscope is now providing data concerning the fabic, diagenesis and weathering of sediments. Some sediments are fragile and care is needed in sample preparation to avoid the introduction of artefacts. For example, moisture removal is often accompanied by dimensional change. Freeze drying and critical point drying appear to reduce this problem. Quantitative methods of fabric and texture analysis are being developed, the data being processed by computer. The Fourier method is able to provide quantitative information concerning shape and texture of particles. This sort of information has been used in studies of provenance, correlation, pedogenesis and environmental processes. Examples are given of the use in engineering of fabric and textural data obtained by the Fourier method and by microscopy.     

基于单次料层冲击破碎质量模型的球磨选择函数

为提高球磨破碎效率,以及构建球磨装置的颗粒破碎过程模型,开展了基于单次料层冲击破碎质量模型的球磨选择函数研究。根据料层夹持形态与落球冲击试验,将料层破碎分为低冲击能下的未完全破碎和高冲击能量下的完全破碎两种状态。分析了单位质量能耗特性和细颗粒对夹持破碎质量的影响,分别建立了两种破碎状态下的料层冲击破碎质量模型。结合冲击能量谱,构建了基于单次料层冲击破碎质量模型的球磨选择函数,将介质球径、颗粒特性等影响球磨破碎效率的参数纳入到选择函数中。     

多层次信息融合在铁谱图像磨粒识别中的应用

针对铁谱图像磨粒识别中异类信息综合利用率较低的问题,提出多层次信息融合的铁谱图像磨粒识别方法。首先,在铁谱图像二值化分割的基础上进行二值滤波,结合彩色铁谱图的R、G、B三分量,实现铁谱图像的彩色滤波。其次,以实际采集的磨粒图像样本为例,提取滤波后二值图像的形态特征,以及滤波后彩色图像的颜色特征;在特征层利用PCA对异类特征进行维数约简,并结合SVM和k-fold交叉验证,实现形态特征和颜色特征的特征层融合;在决策层将异类特征的SVM概率输出结果作为D-S证据理论的基本概率分配函数,实现形态特征和颜色特征的决策层融合。通过与形态学滤波结果对比,验证了本文提出滤波方法的优越性;其次,不同层次的信息融合结果表明,与单独使用颜色特征和形态特征相比,异类信息融合后可实现优势互补,有效提高故障磨粒的识别准确率。