Advanced digital image analysis method dedicated to the characterization of the morphology of filamentous fungus

Filamentous fungi have a complex morphology that induces fermentation process development issues, as a consequence of viscosity increase and diffusion limitations. In order to better understand the relationship between viscosity changes and fungus morphology during fermentations of Trichoderma reesei, an accurate image analysis method has been developed to provide quantitative and representative data for morphological analysis. This method consisted of a new algorithm called FACE that allowed sharp images to be created at all positions, segmentation of fungus, and morphological analysis using skeleton and topological approaches. It was applied and validated by characterizing samples of an industrial strain of Trichoderma reesei that had or had not been exposed to an extreme shear stress. This method allowed many morphological characteristics to be identified, among which nine relevant criteria were extracted, regarding the impact of shear stress on the fungus and on the viscosity of the fermentation medium.     

同态滤波法抑制离轴数字全息零级项

针对离轴数字全息图受记录器件像元尺寸的限制而在数字再现时存在再现像受零级项串扰的问题,提出了一种抑制离轴数字全息零级项的方法.根据图像灰度可由图像照度及表面反射率共同决定的原理,将全息图看作入射分量和反射分量的乘积,利用入射分量变化缓慢且集中在低频段,反射分量反映图像细节并集中在高频段的特性,用同态滤波方法处理数字全息图.设计了同态滤波器,其上下限分别为0.001和1,直径为300 pixel.用该方法实现了对衍射距离为34 cm的菲涅尔数字全息图的零级项抑制,并对重构出的物光对比度以及物体的细节信息进行了增强.     

基于数字图像处理的微光电视像质改善

针对微光电视图像像质缺陷,提出了基于数字图像处理的解决方法。讨论了对微光电视图像进行数字化处理的基本要求,论述了微光电视图像的像质缺陷来源和特点。针对微光电视图像的噪声污染现象,给出了用两种适用实时处理的空间域去除噪声方法。采用直方图均衡化的灰度变换方法来克服微光电视图像显视动态范围偏低的缺点,并通过编程得到了可行有益的微光电视图像的像质改善效果。     

图像采集卡对光学仪器性能数字化检测的影响研究

从介绍图像采集卡在光学仪器性能检测过程中的作用出发 ,详细分析了图像采集卡影响检测结果可信度的若干因素 ,并给出了一个图像采集卡的应用实例     

Three-dimensional morphometry in scanning electron microscopy: a technique for accurate dimensional and angular measurements of microstructures using stereopaired digitized images and digital image analysis

A method for accurate dimensional and angular measurements of microstructures analysed in the scanning electron microscope is described. The method considers central and parallel projections and involves (a) digital image acquisition of stereopaired images from the scanning electron microscope's photodisplay, (b) generation of 3D-image representations, (c) setting of measuring points in the digitized stereopaired images, (d) computation of exact space coordinates ( x / y / z ) from the corresponding point coordinates ( x _L/ y _L; x _R/ y _R), (e) determination of distances and angles between consecutive corresponding points using vector equations, and (f) transfer of computed data into spreadsheets of the data analysis software using dynamic data exchange with simultaneous graphical display of the frequency distribution of variables.
Measurements performed on specimens with known dimensions (grid with 10 μm wide square meshes, polystyrene beads with 0.33 μm diameter) and angles (synthetic crystals of K(Al,Cr)[SO₄], CuSO₄.5H₂O and NaCl) revealed a high accuracy in dimensional as well as angular measurements (total error 1 ± 0.5%).
In Monte Carlo experiments the overall error was found to depend strongly on the size of the measured structure relative to the size of the measurement field (field width).     

浅析数码显微镜分辨率的影响因素

从奈奎斯特采样定理、光源两方面分析对数码显微图像分辨率的影响,表明在物镜确定的情况下,需要选择合适的摄像头、摄像接头、光源、滤色片等,以满足采样定理,有效发挥物镜的分辨率,准确重建出数字图像。     

A semiautomated image analysis procedure for the quantification of dental microwear II

Paleontologists often examine microscopic scratches and pits that form on teeth for indications of the diets of past animals. Most researchers count and measure scratches and pits from photomicrographs to identify microwear patterns for comparison among samples. This paper describes an affordable, available semiautomated image analysis procedure for microwear quantification. An image is downloaded from a scanning electron microscope frame buffer to a microcomputer, and the user identifies microwear features with a mouse-driven pointer. Microwear feature density, dimensions, and orientations are then computed and stored as ASCII files for subsequent data analysis.     

Determination of ZrO2 distribution in an Al2O3 matrix by digital image analysis and point-pattern analysis

Digital image analysis and point-pattern analysis were applied to the microstructures of Al₂O₃-ZrO₂ composites in order to evaluate and classify the distribution of ZrO₂ particles. Scanning electron microscopy micrographs of Al₂O₃-ZrO₂ microstructures were digitized and processed into binary images. Point-pattern analysis was performed using data for mass centers for each ZrO₂ particle and the calculated distribution functions were compared with defined point-pattern distributions—clustered, random, and ordered. Distribution functions for analyzed samples were classified into random groups with different degrees of homogeneity, arising due to the preparation routine. It was shown that the point-pattern analysis can be used to distinguish between the type of distribution and degree of homogeneity for Al₂O₃ ceramic composites.     

Sequential processing of quantitative phase images for the study of cell behaviour in real‐time digital holographic microscopy

Transmitted light holographic microscopy is particularly used for quantitative phase imaging of transparent microscopic objects such as living cells. The study of the cell is based on extraction of the dynamic data on cell behaviour from the time‐lapse sequence of the phase images. However, the phase images are affected by the phase aberrations that make the analysis particularly difficult. This is because the phase deformation is prone to change during long‐term experiments. Here, we present a novel algorithm for sequential processing of living cells phase images in a time‐lapse sequence. The algorithm compensates for the deformation of a phase image using weighted least‐squares surface fitting. Moreover, it identifies and segments the individual cells in the phase image. All these procedures are performed automatically and applied immediately after obtaining every single phase image. This property of the algorithm is important for real‐time cell quantitative phase imaging and instantaneous control of the course of the experiment by playback of the recorded sequence up to actual time. Such operator's intervention is a forerunner of process automation derived from image analysis. The efficiency of the propounded algorithm is demonstrated on images of rat fibrosarcoma cells using an off‐axis holographic microscope.     

Technical note: A toy as tool: a low-cost image analysis system for the evaluation of tumor size in experimental small animal models

Image analysis systems are an essential tool in measurements of size of intraparenchymal tumors or lesions in experimental small animal models. Conventional image analysis systems are relatively expensive. We therefore compared the performance of a professional image analysis system with an inexpensive setup by evaluating tumor size in an orthotopic glioma mouse model. The maximum cross-sectional tumor area of H&E stained brain-slides of two groups of mice (treatment and control group) was measured by two independent investigators using a professional image analysis system (Leica DM IRB microscope) with the Leica Quantimet 500c software, and a low-cost-system (Intel QX3 microscope) with a non-commercial image analysis software. Mean tumor volumes were calculated and the results from each of the image analysis systems, investigators, and treatment effects were compared. The tumor volumes as measured with the low-cost and the professional system differed between -3.7 and +7.5% (P = 0.69-0.99). Measurements made by investigator A and B differed between -7.0 and +3.9% (P = 0.69-0.88). Treatment in all cases significantly reduced the tumor volume between 58.4 and 62.7% (P = 0.0002 or 0.0003), regardless of the investigator or the used image analysis system. We therefore conclude that the QX3 low-cost microscope in combination with a non-commercial image-analysis software represents an inexpensive solution to reliably analyze the size of regions of interest, if they provide a sufficient contrast. However, the low-cost setup due to its low resolution definitely limits a detailed analysis of histologic features.     

Centred contact density functions for the statistical analysis of random sets A stereological study on benign and malignant glandular tissue using image analysis

Real structures investigated in the material and biological sciences, such as minerals or tissues, can often be reduced to two phases. In a stochastic approach, the components of such binary structures may be considered as the union of grains — random sets implanted with their centres at random points — and their complementary space, which is called the pore space. The simplest stochastic germ-grain model is the Boolean model of random sets, which we use here instrumentally as a null model (reference model) for comparison with our biological material. After a brief review of basic properties of the Boolean model and related statistical methods, we introduce centred contact density functions as a new approach. Empirical contact density functions are estimated from the empirical contact distribution functions with an image analyser by dilation of the grain phase. Theoretical contact density functions are then predicted from a set of image parameters, under the assumption that the Boolean model holds. A centred contact density function is the difference between the estimated and the predicted contact density function. Apart from a random error term, centred contact density functions amount to zero irrespective of the area fraction of the grain phase, when the germ-grain model is Boolean. As a section of a spatial Boolean model is a planar Boolean model, the method is also applicable in stereological studies where digitized images are obtained from sections of a three-dimensional structure. Centred contact density functions were determined for mastopathic tissue as compared to mammary cancer, and for tumour-free prostatic tissue as compared to prostatic cancer. For each category of specimens, twenty cases with 10 images each were analysed. Benign and malignant glandular tissue of the aforementioned types deviates significantly from the Boolean model. Centred contact density functions show that malignant transformation is connected with profound geometric remodelling of the pore space.     

Multivariate statistical methods for the analysis of microscope image series: applications in materials science

Multivariate data sets are now produced in several types of microscopy. Multivariate statistical methods are necessary in order to extract the useful information contained in such (image or spectrum) series. In this review, linear and nonlinear multivariate methods are described and illustrated with examples related both to the segmentation of microanalytical maps and to the study of variability in the images of unit cells in high-resolution transmission electron microscopy. Concerning linear multivariate statistical analysis, emphasis is put on the need to go beyond the classical orthogonal decomposition already routinely performed through principal components analysis or correspondence analysis. It is shown that oblique analysis is often necessary when quantitative results are expected. Concerning nonlinear multivariate analysis, several methods are first described for performing the mapping of data from a high-dimensional space to a space of lower dimensionality. Then, automatic classification methods are described. These methods, which range from classical methods (hard and fuzzy C-means) to neural networks through clustering methods which do not make assumptions concerning the shape of classes, can be used for multivariate image segmentation and image classification and averaging.     

Three-dimensional morphometrical study of dendritic spines of the granule cell in the rat dentate gyrus with HVEM stereo images

Number, length, and diameters of dendritic spines of the granule cell in the dorsal leaf of the rat dentate gyrus were measured by using high-voltage electron microscope stereo images of 5-m?m-thick Golgi preparations with the aid of a three-dimensional image analyzer system. Spine densities of 2.02 ± 0.28, 2.28 ± 0.33, and 3.36± 0.35 per 1 μm at distal, middle, and proximal portions of the dendrite were obtained. These values were about 1.6-fold of the previous light microscopical report. Mean three-dimensional spine length were 1.244 ± 0.506 μm, 1.262 ± 0.563 μm, and 1.254 ± 0.584 μm at distal, middle, and proximal portions, respectively, which were about 1.4 times longer than those measured in two dimensions. By using measured morphometrical parameters of spines such as lengths, diameters, and population densities, total spine surface areas of 2.401 μm², 2.806 μm², and 4.180 μm² per 1 μm of the dendrite at distal, middle, and proximal portions, respectively, were obtained. The total surface area of dendrite was about doubled by the addition of the spines at each dendritic portion. The advantageous features and the problems of the present method are discussed.     

A simple 2D composite image analysis technique for the crystal growth study of L‐ascorbic acid

This work was destined for 2D crystal growth studies of L‐ascorbic acid using the composite image analysis technique. Growth experiments on the L‐ascorbic acid crystals were carried out by standard (optical) microscopy, laser diffraction analysis, and composite image analysis. For image analysis, the growth of L‐ascorbic acid crystals was captured as digital 2D RGB images, which were then processed to composite images. After processing, the crystal boundaries emerged as white lines against the black (cancelled) background. The crystal boundaries were well differentiated by peaks in the intensity graphs generated for the composite images. The lengths of crystal boundaries measured from the intensity graphs of composite images were in good agreement (correlation coefficient “r” = 0.99) with the lengths measured by standard microscopy. On the contrary, the lengths measured by laser diffraction were poorly correlated with both techniques. Therefore, the composite image analysis can replace the standard microscopy technique for the crystal growth studies of L‐ascorbic acid.     

Proteome analysis in the study of the bacterial heat-shock response

In recent years, it has become clear that, in addition to the regulation of the expression of specific genes, there are global regulatory systems that control the simultaneous expression of a large number of genes in response to a variety of environmental stresses. The first of these global control systems, and of substantial importance, is the heat-shock response. The heat-shock response is characterized by the induction of a large set of proteins (heat-shock proteins-HSPs) upon shifts to higher temperature and upon exposure to conditions in which proteins are denatured (i.e., alcohols, heavy metals). The heat-shock response is universal and many of the heat-shock proteins are highly conserved among species. In bacteria, the heat-shock response has been studied extensively in several Gram-positive bacteria (Bacillus subtilis) and in the Gram-negative bacteria (i.e., Escherichia coli, Agrobacterium tumefaciens). The first recognition of the molecular abundance of the bacterial heat-shock proteins took place with the introduction of high-resolution two-dimensional polyacrylamide gels (2D gels) to analyze complex mixtures of cellular proteins. Two-dimensional gels, followed by mass spectrometry, were used to define the heat-shock stimulons in several bacteria, and to study the regulatory elements that control the heat-shock response. Here, we review the heat-shock response and its regulation in bacteria. The review will emphasize the use of proteome analysis in the study of this response, and will point out those open questions that can be investigated with proteomics, including mass spectrometry techniques.     

A new method for the two-dimensional analysis of bone structure in human iliac crest biopsies

A new method is described for the analysis of the two-dimensional structural pattern of trabecular bone in human iliac crest biopsies. 8 μm thick undecalcified sections stained with the von Kossa technique were examined at a magnification of ×9. Using an Ibas II image analyser, the ratio of nodes to free ends and the length of different strut types (cortex to free end, node or cortex, free end to free end and node to node, loop or free end) expressed as a percentage of total strut length were assessed. The reproducibility of the method was good for most of the measured indices but inter-observer and inter-section variation were greater. Comparison of biopsy sections obtained from eleven young healthy control subjects and eleven patients with hepatic osteoporosis revealed a significantly higher node to free end ratio, node to loop and node to node strut length and significantly lower cortex to free end and free end to free end strut length in the controls. No significant differences were seen in node to free end, cortex to cortex or cortex to node strut length. This approach to trabecular bone structure analysis should prove useful in determining patterns of bone loss in health and disease and in examining the effects of treatment on bone structure in osteoporosis.     

Quantitative and reliable in vitro method combining scanning electron microscopy and image analysis for the screening of osteotropic modulators

The increased generation and up-regulated activity of bone resorbing cells (osteoclasts) play a part in the impairment of bone remodeling in many bone diseases. Numerous drugs (bisphosphonates, calcitonin, selective estrogen receptor modulators) have been proposed to inhibit this increased osteoclastic activity. In this report, we describe a pit resorption assay quantified by scanning electron microscopy coupled with image analysis. Total rabbit bone cells with large numbers of osteoclasts were cultured on dentin slices. The whole surface of the dentin slice was scanned and both the number of resorption pits and the total resorbed surface area were measured. Resorption pits appeared at 48 h and increased gradually up to 96 h. Despite the observation of a strong correlation between the total resorption area and the number of pits, we suggest that area measurement is the most relevant marker for osteoclastic activity. Osteotropic factors stimulating or inhibiting osteoclastic activity were used to test the variations in resorption activity as measured with our method. This reproducible and sensitive quantitative method is a valuable tool for screening for osteoclastic inhibitors and, more generally, for investigating bone modulators.     

A study on the development of image analysis instrument and estimation of mass,volume and center of gravity using CT image in Korean

In this study, the body segment parameters of Koreans were estimated using two-dimensional CT images of cadavers. A software instrument was developed in this study for the image analyses. The software converts 2D images into 3D images and reconstructs the full-body image to automatically distinguish and divide it into fat, bone, and muscle tissues, depending on the body components. The body segments were manually separated by joints, and the software automatically calculated the average mass, volume, and center of gravity of the body segment, and the results were used to analyze the relationship between the body segments and the body characteristics and to build a regression model with statistical significance. The regression model can estimate the mass and volume of segments based on the gender, height, and weight. The software instrument was able to rapidly analyze full-body CT images, and it allows users to easily repeat the analysis.     

基于ANSYS的钻井泵泵阀有限元分析

运用ANSYS对BF700I新型钻井泵泵阀的带筋阀座进行了三维有限元分析。结果表明,改进的泵阀阀座具有较好的稳定性;在泵阀小端面接近内锥面处,应力变形最大;阀座外锥面底部边缘的应力较大;中间阀盘的底部轴孔边缘也有很大的应力集中。