Digital image tiles: a method for the processing of large sections

Segmentation of large areas of light microscopic slides into N by N fields, and each of these fields into M digital image tiles, allows the scanning, storage and digital processing of large images. Any of the original N² fields or composites of M adjacent tiles can be recalled to the video display for analysis. Developed procedures for use on a microscope equipped with a precision scanning stage allow registration of the image coordinates (X-Y) for any original or composite field and the alignment of one of these fields along the depth (Z) axis by means of external, machined fiducial marks in serial sections. To facilitate work whenever unavoidable, we have incorporated methods for digital image panning and zooming (changes of magnification) and discuss their use and implications.     

New generation scanning electron microscopy technology based on the concept of active image processing

A new generation of scanning electron microscopy (SEM) technology is proposed based on the concept of “active image processing.” In order to collect sufficient data for a purpose which is defined in the utilization of active image processing, we may need more devices from among a variety of useful hardware, for example, a digital scan generator with meaningful parameters and an analog-to-digital converter for ultrahigh density recording. After the data acquisition, the application of some digital image processing techniques is certainly effective, because the method in question is specially designed so that the property of obtained data will be suitable for the application of these techniques. The present technology should produce a variety of attractive options in the field of SEM.     

Videomicroscopy, image processing, and analysis of whole histologic sections of the human brain

Serial histologic sections of a whole human brain may have extensions of up to 130 x 130 mm within the coronal plane around the temporal lobe. To date, however, technology has not provided a bright field microscope that is able to shift the object holder continuously in the x- and y-direction over such distances and still possess the same optical capabilities as comparable devices. We developed a new light microscope to continuously quantify such sections. We also developed the computing environment for controlling the device and for analyzing the data produced. In principle, we are now able to quantify each neuron of a human brain. The data ultimately will provide the most detailed structural information about the human brain ascertained thus far. Such detailed information of the spatial distribution of neurons is essential to develop realistic models for simulation of large-scale neuronal networks and to investigate the significance of neuronal arrangements with respect to neuronal signal processing in the CNS. After preprocessing of the data produced by the new microscope, we are able to detect lamination patterns in the spatial distribution of gravity centers of cells. Furthermore, morphological features like size of the projection area and mean staining intensity are visualized as a particle process. The particle process presents the sizes and staining intensity of perikaryons and allows a distinction of gray matter and white matter. These results provide evidence that the system works correctly and can be applied to a systematic analysis of a larger sequence of serial histologic sections. The objective of this study is to introduce the very large section analyzing microscope (VLSAM) and to present the initial data produced by the system. Moreover, we will discuss workload and future developments of the parallel image analysis system that are associated with the microscope.     

Confocal microscopy and image processing in the study of plant nuclear structure

We describe measurements of the point spread function (PSF) for a confocal microscope and compare them with the PSF for a conventional (wide-field) fluorescence microscope. In situ hybridization with probes to telomere and ribosomal rDNA sequences, combined with three-dimensional (3-D) microscopy, has been used to study interphase nuclei in root tissue of Pisum sativum and Vicia faba. Nearly all the telomeres in both species are located at the nuclear envelope, and are highly clustered in the Vicia tissues, suggesting specific binding interactions. rDNA labelling in P. sativum shows four brightly staining knobs, corresponding to condensed regions of the rDNA genes from the two pairs of nucleolar organizer genes in this species, arranged approximately tetrahedrally around each nucleolus. Deconvolution using the measured PSFs can be used to improve these images, revealing a fibrous substructure in the perinucleolar knobs, and a large amount of interconnecting internal structure, which we suggest represents rDNA both in the fibrillar centres and also more diffuse, widely dispersed rDNA. Finally we show that accurate conventional data coupled with deconvolution can produce 3-D reconstructions comparable to those obtainable with confocal microscopy, but that the clearest images are obtained by applying deconvolution to the confocal data.     

基于图像处理的矿用乳化液浓度在线检测

针对传统矿用乳化液浓度检测方法的需人工取样、有滞后性、测量精度低等普遍存在的问题,对光入射到乳化液后的成像进行了研究,提出了一种基于光全反射原理的专用光学检测系统。对成像进行了几何建模,研究了其特性参数与乳化液浓度的关系,采用建立的函数关系对各种不同浓度的乳化液进行了检测。实验结果表明,基于图像处理的在线检测法检测精度可达0.001%,相比于传统检测方法的检测精度0.5%,具有更高的精度和可靠性。该检测方案克服了传统检测方法无法实现在线检测的缺点,具有很好的实用性。     

Three-dimensional reconstruction of a plant dictyosome from series of ultrathin sections using computer image processing

A single dictyosome from an actively secreting ovary gland cell of Aptenia cordifolia has been reconstructed in 3-D from a series of twenty-nine electron micrographs by computer image processing. The reconstruction is presented under different viewing angles in the form of shaded perspective displays. From these displays the entire dictyosome, surrounded by numerous vesicles, appears to be more a spherical than a flat body. The plate-like region of the dictyósome is demonstrated when only a portion of the electron micrographs is used for the image processing, leading to ‘cut-off’ displays. Since some upper planes were removed, such ‘cut-off’ displays revealed both tubular connections between cisternae of the dictyosome and the neighbouring endoplasmic reticulum as well as tubular continuities between adjacent Golgi cisternae within the same stack. Possible consequences of both types of interconnections on transport and processing of proteins and glycoproteins are discussed.     

Three-dimensional electron microscopy of entire cells

The digital processing of serial electron-microscope sections containing laser-induced topographical references allows a three-dimensional (3-D) reconstruction of entire cells at a depth resolution of 40–60 nm by the use of novel image analysis methods. The images are directly processed by a video-camera placed under the electron microscope in TEM mode or by the electron counting device in STEM mode. The deformations associated with the cutting of embedded cells are back-calculated by new computer algorithms developed for image analysis and treatment. They correct the artefacts caused by serial sectioning and automatically reconstruct the third dimension of the cells. Used in such a way, our data provide definitive information on the 3-D architecture of cells. This computer-assisted 3-D analysis represents a new tool for the documentation and analysis of cell ultrastructure and for morphometric studies. Furthermore, it is now possible for the observer to view the contents of the reconstructed tissue volume in a variety of different ways using computer-aided display techniques.     

面向对象的通用图象处理系统的体系结构研究

在面向对象技术的基础上本文提出一个通用图象处理系统的体系结构。它基于通用图象处理系统的基本功能来组织对象,以图象数据对象为核心构建系统框架。基于文档-视图结构,使用VisualC 实现了通用图象处理系统的一个原型系统,该原型系统由图象文件操作模块、图象预处理模块、图象处理模块、人工调整模块、结果输出模块等五个模块组成。对体系结构的模型分析与原型系统的测试表明,该体系结构对图象处理系统的设计开发是非常有效的。     

基于图像处理的在线实时监测系统设计

针对传统远程监测存在的图像质量差的问题,提出一种基于图像处理的远程实时监测系统.利用Web技术对系统整体架构、系统功能和开发环境等进行搭建,从而为图像的远程监控提供功能处理界面;利用图像处理算法对彩色模型选择、图像归一化、图像分割和图像去噪等进行处理,提高了图像采集的质量.最后通过系统运行,验证了该算法的可行性,为远程图像实时监控系统开发提供参考.     

基于TMS320C6711图像处理系统的开发

该系统实现了一个比较完整的图像采集，传输．计算和显示的硬件实验系统，它是由real-time公司提供的ICETEK-C6711-A板和ICETEK-IDK-YUV板以及彩色监视器．摄像头．各种连线所组成。摄像头所捕捉到的图像通过被编写在DSP上的中值滤波，直方围增强，灰度阈值处理，图象锐化．边缘检测等图像处理程序进行优化后能够实现576^＊720pixel图像的准实时稳定的传输及送显。用户还可以通过CCS在该系统上加载自己的图像处理算法程序．来检验处理的效果。     

基于图像测量技术的微透镜定焦方法研究

简要描述了基于图像处理方法的微透镜阵列,焦距测量方法。该方法采用图像处理技术解决测量中的定焦问题。文中介绍了几种清晰度评价函数。并对其定焦的精度进行比较,选出较优的算法用来判断焦点位置。实验证明使用该方法可以满足微透镜焦距测量精度要求。     

A combined light and electron microscopic method for the visualization of the same in vitro neuron by radioautography and serial sections

We report here on a technical improvement which makes it possible to study, at the ultrastructural level, a dopaminergic neuron which has been previously identified by light microscopy. Primary cultures of virtually pure mesencephalic neurons from mouse embryos were obtained. These cultures were kept for 6 days, then incubated with tritiated dopamine. fixed and embedded in Epon. The dopaminergic neurons were firstly visualized by radioautography directly through Epon blocks in toto by light microscopy. In a second step, ultrathin sections of the identified dopaminergic cells were prepared and the neurons observed at the electron microscopy level. The dopaminergic nature of these neurons was regularly checked by radioautographic control on some selected ultrathin sections.     

Determination of the physical structure of biological materials at biosensor interfaces by techniques of increasing magnification from microscopic to molecular scale

Chemical selectivity of biosensors is derived from biological materials interfaced to the surface of transducing devices. Molecular recognition events lead to macroscopic function suitable for analytical measurements. The structure-function relationships of biochemical species at interfaces must be established to characterize and optimize biosensor operation. The techniques of ellipsometry, fluorescence microscopy, electron microscopy, and scanning tunneling microscopy are used to investigate the structure of monolayers and multilayers of proteins and lipids at interfaces that are prepared by Langmuir-Blodgett techniques and by self-assembly from bulk solution. The relative merits and limitations of the measurement techniques in the determination of aspects of interfacial structure are considered.     

Imaging of submicron objects with the light microscope

Using conventional optics coupled with image processing, several submicron objects have been studied with light microscopy. These include polystyrene beads (88, 264 and 557 nm), frustules from the diatom Pleurosigma angulatum and the T-4 bacteriophage, either attached to its host, Escherichia coli, or free in the medium. The best results were obtained from dark-field and phase contrast optics. Digital image processing with the use of colour look-up tables in real time greatly promoted precise focusing of the objects. Selective discrimination of image histogram grey values allowed for selection of a sharp contour boundary, thus more effectively delimiting the submicron objects. User interactive scaling of the diffraction limited boundaries greatly improved discrimination of the objects. While Abbe limits have not been surpassed by these techniques, the greater ‘apparent resolution’ could be attributable to a ‘better recognition’ of the submicron objects examined. Equivalence of images of polystyrene beads and bacteriophages was demonstrated with light and electron microscopy of the same field.     

正交设计方法在优化空瓶检测图像处理参数中应用

介绍空瓶检测的图像处理关键技术,对酒瓶破损模式、空瓶检测方法的特点进行分析.以瓶身检测为例,用三水平三因素的正交设计对空瓶检测的图像处理参数,即Sobel、Prewitt、Roberts等边缘检测算子、处理步长以及决策算法等进行组合优化设计.通过对试验结果和极差值的分析,得出了各参数对空瓶检测精度和实时性影响的主次因素,为优化参数、平衡空瓶检测的准确率与工作时间提供了依据.在与论文实验条件相类似的情况下,推荐采用Prewitt、处理步长为1、加权求和决策算法的算法组合.本方法可推广到不同检测系统要求下所确定的最优参数组合中.     

应用数字图像处理技术检测塔式起重机钢丝绳的损伤

介绍1种采用数字图像处理实现塔式起重机钢丝绳损伤检测的新方法,对采集到的钢丝绳图像进行预处理以减少噪声的污染和影响,根据损伤钢丝绳的特征对预处理的图像进行分割,对图像进行数学形态学处理并判断钢丝绳是否存在损伤。     

Simple three-dimensional imaging of HRP-labelled neurons with the aid of an image processor

A method is described for visualizing the three-dimensional structure of horseradish peroxidase-labelled neurons in the central nervous system of Xenopus laevis tadpoles. The reconstruction is simple, and performed on a commercially available image processing system. Minimal operational skill is required for using this method. Also, various improvements were made where user's simplicity (user-friendliness), speed and required computer memory are concerned. In addition, the excellent properties of the IBAS II system, such as image filtering and image editing functions are fully available, offering a fast, clean and complete final image reconstruction.     

Application of recording and processing of energy-filtered image sequences for the elemental mapping of biological specimens: Imaging-Spectrum

Computerized energy-filtered transmission electron microscope (EFTEM) permits the recording and the processing of energy-filtered images, allowing a part of an electron energy-loss spectrum for each picture element to be obtained. This method, called ‘Imaging-Spectrum’, uses a Zeiss CEM902 coupled to several image analysis systems. The actual configuration records sequences of 48 images, 256 × 256 pixels, in steps of the energy loss, ΔE. Processing these sequences results in part of a core-loss EELS-spectrum for each pixel. This approach produces elemental maps with a short processing time. We have implemented three kinds of background calculation for the image subtraction. The influence of the irradiation dose and of the energy selecting slit width on the quality of the spectra is investigated. The method is applied to the analysis of some biological specimens (pericellular coat behaviour during adhesion between macrophages and red blood cells and location of calcite microcrystals in dental pulp cells). The Imaging-Spectrum method appears to be suitable for the analysis of large areas.     

Measurement of the geometric features of a cutting tool edge with the aid of a digital image processing technique

For the further development of new machining systems such as flexible manufacturing systems (FMS), this paper reports on a new measuring system developed for the multi-purpose understanding of the geometric features of cutting tool edges having serious influences on the machining outputs. In this system, to detect three-dimensional (3D) features of the toolface and tool flank only from the two-dimensional (2D) microscopic observation of the toolface and to describe an overall 3D geometric feature of the tooledge as a digital 2D image as well as to enable inexpensive system development, two convenient measuring principles are used. These are the scanning optical section method for the toolface contour mapping and the cutting edge profile comparison method for the flank wear landwidth estimation. Based on these, experimental hardware was arranged and various processing softwares were developed. They were applied to an observation of the cutting edge wear development in some cylindrical turning experiments, to show that various effective cutting edge parameters could be easily extracted from the integrated toolface image recorded, and that sufficient accuracy and resolution for practical use could be obtained.