Assessment and Visualization of Ki67 Heterogeneity in Breast Cancers through Digital Image Analysis

The Ki67 index (KI) is a standard clinical marker for tumor proliferation;however, its application is hindered by intratumoral heterogeneity. In this study, we used digital image analysis to comprehensively analyze Ki67 heterogeneity and distribution patterns in breast carcinoma. Using Smart Pathology software, we digitized and analyzed 42 excised breast carcinoma Ki67 slides. Boxplots, histograms, and heat maps were generated to illustrate the KI distribution. We found that 30% of cases (13/42) exhibited discrepancies between global and hotspot KI when using a 14% KI threshold for classification. Patients with higher global or hotspot KI values displayed greater heterogenicity. Ki67 distribution patterns were categorized as randomly distributed (52%, 22/42), peripheral (43%, 18/42), and centered (5%, 2/42). Our sampling simulator indicated analyzing more than 10 high-power fields was typically required to accurately estimate global KI, with sampling size being correlated with heterogeneity. In conclusion, using digital image analysis in whole-slide images allows for comprehensive Ki67 profile assessment, shedding light on heterogeneity and distribution patterns. This spatial information can facilitate KI surveys of breast cancer and other malignancies.

<i>In Vitro</i>Bioassay of Allelopathy in Four Bamboo Species;<i>Bambusa multiplex, Phyllostachys bambusoides, P. nigra, Sasa kurilensis</i>, Using Sandwich Method and Protoplast Co-Culture Method with Digital Image Analysis

Moderately strong allelopathic activities were found in four bamboo species, Bambusa multiplex cv. Houraichiku;Phyllostachys bambusoides cv. Madake;P. nigra cv. Hachiku;Sasa kurilensis cv. Chishimazasa, which are of different classification or of different ecological distributions, using the “Sandwich Method”, which assays the dried leaves on growth of lettuce seedlings. Only small difference of activity was found among the four bamboo species. In addition, “Protoplast Co-culture Method” for assay of allelopathy in a 50 μL liquid medium using a 96 well culture plate, was applied to the suspension cultures of the four bamboo species. Protoplasts were isolated from two-week cultured suspension cells of four bamboo species using Cellulase RS and Pectolyase Y-23 in 0.6 M mannitol. At low protoplast densities of bamboo, B. multiplex and P. bambusoides stimulated the recipient lettuce growth, i.e., non-spherically cell enlargement and cell divisions observed under an inverted microscope, while protoplasts of P. nigra and S. kurilensis were less stimulatory or inhibitory. Inhibitory effect of S. kurilensis was the strongest among four bamboo species. Furthermore, highly inhibitory effects of S. kurilensis protoplasts on yellow color accumulation of lettuce protoplasts were clearly observed by analysis of a scanned digital image of a 96-well culture plate. Differences and causes of the allelopathic activities were discussed comparing with other plant species studied using the same assay methods.

Collagen proportionate area of liver tissue determined by digital image analysis in patients with HBV-related decompensated cirrhosis

BACKGROUND: The accurate assessment of the degree of hepatic fibrosis plays a critical role in guiding the diagnosis, treatment and prognostic assessment of chronic liver diseases. Liver biopsy is currently the most reliable method to evaluate the severity of hepatic fibrosis. However, liver biopsy is an invasive procedure associated with morbidity and mortality, and has several limitations in patients with decompensated cirrhosis. There is no report on the collagen proportionate area (CPA) of liver tissue in the decompensated stage of cirrhosis. This study aimed to determine the CPA of resected liver tissue samples from patients with HBV-related decompensated cirrhosis using digital image analysis, and to analyze the relationship between the CPA and liver functional reserve. METHODS: Fifty-three resected liver tissue samples from liver transplant patients with chronic hepatitis B-induced decompensated cirrhosis were stained with Masson’s trichrome, and the CPA in these samples was quantitatively determined using digital image analysis. The values of relevant liver function just before liver transplantation, the CPA in liver tissue, and their correlation were analyzed. RESULTS: The mean CPA at the decompensated stage of cirrhosis was 35.93±14.42% (11.24%-63.41%). The correlation coefficients of the CPA with a model for end-stage liver disease score, serum total bilirubin and international standard ratio of prothrombin B were 0.553, 0.519 and 0.533, respectively (P<0.001). With increasing CPA values, the three indices reflecting liver functional reserve also changed significantly.CONCLUSIONS: The degree of fibrosis may be correlated with the functional reserve. With the advancement of fibrosis, the liver functional reserve is attenuated accordingly.

In-situ strain measurement and error analysis of arc welding with 2D digital image correlation

The deformation and residual stress generated by the welding process can seriously affect the use of components.As a result,it is very important to understand the evolution of stress and strain during the welding process.The strain measurement method based on digital image correlation(DIC)is an excellent method to detect welding strain and residual stress.The out-of-plane translation and out-of-plane rotation introduce errors to the two-dimensional DIC.In this paper,the causes of errors are analyzed theoretically,and the formulas of errors caused by the out-of-plane displacement and the out-of-plane rotation are derived.The out-of-plane translation experiment and the out-of-plane rotation experiment were carried out to verify the theory,and the experimental results are consistent with the theoretical analysis results.The error caused by the out-of-plane translation can be reduced by increasing the object distance;the error caused by the out-of-plane rotation is greatly affected by the rotation angle.

DIGITAL IMAGE ANALYSIS OF PHASE STRUCTURE IN THE BLEND OF POLY(BUTYLENE TEREPHTHALATE-COCAPROLACTONE) WITH POLYCARBONATE

The digital image processing technique was used for the extraction of physical information from the temporal image of phase structure in the process of phase separation in the blend of poly (butylene terephthalate-co-caprolactone )with polycarbonate. The power spectrum images of the two-dimensional Fourier transformation (2DFT) of the temporal image of phase structure reflect the change of the phase size with time.

Automatic Computer Analysis of Digital Images of Triple-Antibody-Stained Prostate Biopsies

Background: Worldwide, prostatic adenocarcinoma is the most common tumour type among men. Aim: The aim of the present investigation was to develop a computer program to identify normal prostate biopsies and distinguish them from biopsies showing premalignant alterations (LGPIN, HGPIN) and adenocarcinoma. Method: Prostate biopsies (n = 2094) taken from 191 consecutive men during 2016 were stained with triple immunehistochemisty (antibodies to AMACRA, p63 and CK 5). Digital images of the biopsies were obtained with a scanning microscope and used to develop an automatic computer program (CelldaTM), intended to identify the morphological alterations. Visual microscopic finding was used as a reference. Result: Of the 191 men, 121 (63.4%) were diagnosed as having prostate adenocarcinoma and 70 (36.6%) as having no malignancy on the basis of the visual microscopy. In comparison, computer analysis identified 134 (70.2%) men with malignant disease and 57 (29.8%) with non-malignant disease after exclusion of artifacts, which constituted 10.4% of areas (indicated as malignant disease). Discrepant results were recorded in 15 (7.9%) men, and in 14 of these cases, HGPIN and areas suggestive of early invasion were common. Thus, it was uncertain whether these cases should be regarded as malignant or not. The agreement between the visual examination and the computer analysis was 92.1% (kappa value 0.823, sensitivity 99.2 and specificity was 0.80). Conclusion: It seems that computer analysis could serve as an adjunct to simplify and shorten the diagnostic procedure, first of all by ensuring that normal prostate biopsies are sorted out from those sent for visual microscopic evaluation.

Automated Detection and Quantification of Prostate Cancer in Needle Biopsies by Digital Image Analysis

Introduction: Triple immunohistochemical (IHC) stains including antibodies specific for alpha-methylacyl-CoA-racemase and basal cell markers have been a valuable aid in accurate identification of prostate carcinoma. However, accurate quantification of minuscule areas of prostate carcinoma in biopsy specimens can often be a challenge. Here we assessed the diagnostic value and quantitative use of automated digital image analysis on triple IHC stained prostate needle biopsies. Methods: Twelve cases of prostate needle biopsy material including 75 needle cores were stained with triple-antibody cocktail (P504S + 34βE12 + p63). Slides were digitally scanned with the APERIO digital image analyzer and evaluated with the GENIE pattern and color recognition digital image analysis that we developed. A slide with known areas of adenocarcinoma, high grade prostatic intraepithelial neoplasia (PIN), benign glands and stroma was used as a training set for the automated digital image analysis platform. Results: Among 75 needle biopsy cores, 19 (25.33%) contained adenocarcinoma by histology. Digital image analysis recognized adenocarcinoma in 95% of these needle biopsies. The average area of the needle biopsy was 7.63 mm2 and overall the average area of tumor was 0.196 mm2. The smallest area of tumor recognized by the program was 0.0022 mm2 (0.0363% of the core) and the largest was 0.62 mm2 (8.17% of the core) among needle core biopsies. False positives resulted from areas of high grade PIN with patchy basal cells. The false negative was caused by uneven AMACR staining in one area of adenocarcinoma. Digital recognition of areas of interest was improved by three successive image analysis training which increased the sensitivity and specificity from 83% and 89% to 90% and 93%, respectively. Conclusions: Digital image analysis in concert with IHC triple staining may be useful for accurate detection and quantitative analysis of small foci of prostatic adenocarcinoma. Defining methods to increase the sensitivity and specificity of quantitative automated digital image analysis will likely evolve as an area of investigation. Future automated digital scanning and innovative pattern and color recognition technologies may open avenues for classifying a variety of prostate lesions.

Impact of blurs on machine-learning aided digital pathology image analysis

BACKGROUND Digital pathology image(DPI)analysis has been developed by machine learning(ML)techniques.However,little attention has been paid to the reproducibility of ML-based histological classification in heterochronously obtained DPIs of the same hematoxylin and eosin(HE)slide.AIM To elucidate the frequency and preventable causes of discordant classification results of DPI analysis using ML for the heterochronously obtained DPIs.METHODS We created paired DPIs by scanning 298 HE stained slides containing 584 tissues twice with a virtual slide scanner.The paired DPIs were analyzed by our MLaided classification model.We defined non-flipped and flipped groups as the paired DPIs with concordant and discordant classification results,respectively.We compared differences in color and blur between the non-flipped and flipped groups by L1-norm and a blur index,respectively.RESULTS We observed discordant classification results in 23.1%of the paired DPIs obtained by two independent scans of the same microscope slide.We detected no significant difference in the L1-norm of each color channel between the two groups;however,the flipped group showed a significantly higher blur index than the non-flipped group.CONCLUSION Our results suggest that differences in the blur-not the color-of the paired DPIs may cause discordant classification results.An ML-aided classification model for DPI should be tested for this potential cause of the reduced reproducibility of the model.In a future study,a slide scanner and/or a preprocessing method of minimizing DPI blur should be developed.

Simulation of Fracture Process of Lightweight Aggregate Concrete Based on Digital Image Processing Technology

The mechanical properties and failure mechanism of lightweight aggregate concrete(LWAC)is a hot topic in the engineering field,and the relationship between its microstructure and macroscopic mechanical properties is also a frontier research topic in the academic field.In this study,the image processing technology is used to establish a micro-structure model of lightweight aggregate concrete.Through the information extraction and processing of the section image of actual light aggregate concrete specimens,the mesostructural model of light aggregate concrete with real aggregate characteristics is established.The numerical simulation of uniaxial tensile test,uniaxial compression test and three-point bending test of lightweight aggregate concrete are carried out using a new finite element method-the base force element method respectively.Firstly,the image processing technology is used to produce beam specimens,uniaxial compression specimens and uniaxial tensile specimens of light aggregate concrete,which can better simulate the aggregate shape and random distribution of real light aggregate concrete.Secondly,the three-point bending test is numerically simulated.Thirdly,the uniaxial compression specimen generated by image processing technology is numerically simulated.Fourth,the uniaxial tensile specimen generated by image processing technology is numerically simulated.The mechanical behavior and damage mode of the specimen during loading were analyzed.The results of numerical simulation are compared and analyzed with those of relevant experiments.The feasibility and correctness of the micromodel established in this study for analyzing the micromechanics of lightweight aggregate concrete materials are verified.Image processing technology has a broad application prospect in the field of concrete mesoscopic damage analysis.

Laboratory visualization of damage asymmetry formation of rock fracture via acoustic emission and digital imaging correlation

Rock fracture mechanics and accurate characterization of rock fracture are crucial for understanding a variety of phenomena interested in geological engineering and geoscience.These phenomena range from very large-scale asymmetrical fault structures to the scale of engineering projects and laboratory-scale rock fracture tests.Comprehensive study can involve mechanical modeling,site or post-mortem investigations,and inspection on the point cloud of the source locations in the form of earthquake,microseismicity,or acoustic emission.This study presents a comprehensive data analysis on characterizing the forming of the asymmetrical damage zone around a laboratory mixed-mode rock fracture.We substantiate the presence of asymmetrical damage through qualitative analysis and demonstrate that measurement uncertainties cannot solely explain the observed asymmetry.The implications of this demonstration can be manifold.On a larger scale,it solidifies a mechanical model used for explaining the contribution of aseismic mechanisms to asymmetrical fault structures.On a laboratory scale,it exemplifies an alternative approach to understanding the observational difference between the source location and the in situ or post-mortem inspection on the rock fracture path.The mechanical model and the data analysis can be informative to the interpretations of other engineering practices as well,but may face different types of challenges.

Estimation of Ground Deformation Caused by the Earthquake (M7.2) in Japan,2008,from the Geomorphic Image Analysis of High Resolution LiDAR DEMs

In this study, a new method for quantitative and efficient measurement for the ground surface movement was developed. The feature of this technique is to identify geomorphic characteristics by image matching analysis, using the intelligent images made from high resolution DEM(Digital Elevation Model). This method is useful to extract the small ground displacement where the surface shape was not intensely deformed.

On Secure Digital Image Watermarking Techniques

Digital watermarking is used to hide the information inside a signal, which can not be easily extracted by the third party. Its widely used application is copyright protection of digital information. It is different from the encryption in the sense that it allows the user to access, view and interpret the signal but protect the ownership of the content. One of the current research areas is to protect digital watermark inside the information so that ownership of the information cannot be claimed by third party. With a lot of information available on various search engines, to protect the ownership of information is a crucial area of research. In latest years, several digital watermarking techniques are presented based on discrete cosine transform (DCT), discrete wavelets transform (DWT) and discrete fourier transforms (DFT). In this paper, we propose an algorithm for digital image watermarking technique based on singular value decomposition;both of the L and U components are explored for watermarking algorithm. This technique refers to the watermark embedding algorithm and watermark extracting algorithm. The experimental results prove that the quality of the watermarked image is excellent and there is strong resistant against many geometrical attacks.

An image analysis method for quantification of hepatic perfusion based on contrast-enhanced ultrasound imaging

Information about hepatic perfusion is used in clinical liver disease diagnosis. An image analy-sis system can help physicians make efficient and accurate diagnosis. The objective of this study is to propose an image analysis method for the quantification of the hepatic perfusion based on contrast-enhanced ultrasound imaging (CEUI). The proposed method contains frame selection, image registration, digital subtraction and grey-scale calculation. Then, by processing an image sequence, a time-intensity curve (TIC) for hepatic perfusion is derived. The kernel of this image analysis technology is digital subtrac-tion and its accuracy is improved by frame selec-tion and image registration. The advantage of this method is that it can obtain the perfusion information of the whole liver which is rarely ob-tained by traditional image analysis technology;therefore, it is a supplement of the traditional image analysis method. This method is applied on the quantification of a rabbit’s hepatic perfu-sion and the result shows the efficiency of it.

Bubble dynamics properties of B-particles in a quasi-2D gas-solid fluidized bed:Computational particle fluid dynamics numerical simulation and post-processed by digital image analysis technique

Bubble dynamics properties play a crucial and significant role in the design and optimization of gas-solid fluidized beds.In this study,the bubble dynamics properties of four B-particles were investigated in a quasi-two-dimensional(quasi-2D)fluidized bed,including bubble equivalent diameter,bubble size distribution,average bubble density,bubble aspect ratio,bubble hold-up,bed expansion ratio,bubble radial position,and bubble velocity.The studies were performed by computational particle fluid dynamics(CPFD)numerical simulation and post-processed with digital image analysis(DIA)technique,at superficial gas velocities ranging from 2u_(mf) to 7u_(mf).The simulated results shown that the CPFD simulation combining with DIA technique post-processing could be used as a reliable method for simulating bubble dynamics properties in quasi-2D gas-solid fluidized beds.However,it seemed not desirable for the simulation of bubble motion near the air distributor at higher superficial gas velocity from the simulated average bubble density distribution.The superficial gas velocity significantly affected the bubble equivalent diameter and evolution,while it had little influence on bubble size distribution and bubble aspect ratio distribution for the same particles.Both time-averaged bubble hold-up and bed expansion ratio increased with the increase of superficial gas velocity.Two core-annular flow structures could be found in the fluidized bed for all cases.The average bubble rising velocity increased with the increasing bubble equivalent diameter.For bubble lateral movement,the smaller bubbles might be more susceptible,and superficial gas velocity had a little influence on the absolute lateral velocity of bubbles.The simulated results presented a valuable and novel approach for studying bubble dynamics properties.The comprehensive understanding of bubble dynamics behaviors in quasi-2D gas-solid fluidized beds would provide support in the design,operation,and optimization of gas-solid fluidized bed reactors.

Arterial Imaging in Digital Gangrenes Associated with Scleroderma-Spectrum Disorders

Objectives: Digital refractory gangrene is rarely found in collagen diseases, including systemic sclerosis and is possibly caused by similar underlying vascular damage in peripheral arterial disease (PAD) such as arteriosclerosis obliterans (ASO) and/or thromboangiitis obliterans (TAO) by unclarified mechanisms other than vasculitis and thrombosis. This study evaluated the radiological imaging in patients with digital gangrene associated with collagen disease and compared the images with those of PAD based on the results of laboratory and histopathological examinations. Methods: Angiography, MR angiography and/or CT angiography were performed on 6 patients with refractory gangrene or extensive ulcers accompanied by scleroderma-spectrum disorders;3 with diffuse systemic sclerosis, 1 with limited systemic sclerosis, 1 with overlap syndrome and 1 with Sj?gren’s syndrome. Results: Although the vascular alterations in collagen diseases were similar to those in PAD, the abnormal image findings (occlusion or stenosis of the arteries with smooth vessel walls) found in collagen diseases did not include atheromatous plaque, which are worm-like vessels that are characteristic of those observed in PAD. Conclusions: Some cases of digital gangrene seen in collagen diseases show similar vascular imaging patterns to those of PAD and comprehensive examinations including arterial imaging can be useful for the diagnosis of these unrecognized vascular changes other than vasculitis or digital thrombosis.

Mapping Microscale Strain Fields Around a Crack Tip in Molybdenum Via Geometric Phase Analysis and Digital Image Correlation

Uniaxial tension tests were conducted on single-edge-notched tensile specimens of pure molybdenum with a mesh grid pattern in front of the notch. A series of images of crack initialization and propagation with a distorted mesh grid pattern were obtained by means of in situ scanning electron microscopy. Strain fields around the crack tip were mapped successively using geometric phase analysis and digital image correlation techniques, and then compared with the predictions obtained through linear elastic fracture mechanics （LEFM）. The comparison shows that the measured strain distribution ahead of the crack tip is consistent with the LEFM predictions of up to 25 μm from the crack tip.

Designing Thought,Technical Line and Some Theoretical Issues on Geospatial Digital Framework of China

Based on the developing tendency of present China’s basic GIS,this paper discusses the designing idea for scales of 1∶10 000,1∶50 000, 1∶250 000 and 1∶1 000 000 pyramid_like multi_layer and multi_resolution of the basic GIS.A technical line for the construction of basic GIS of the whole country and various provinces for sustainable development is put forward.And some important theoretical GIS issues touched by the technical process are discussed.

An Object-based Approach for Two-level Gully Feature Mapping Using High-resolution DEM and Imagery： A Case Study on Hilly Loess Plateau Region, China

Gully feature mapping is an indispensable prerequisite for the motioning and control of gully erosion which is a widespread natural hazard. The increasing availability of high-resolution Digital Elevation Model(DEM) and remote sensing imagery, combined with developed object-based methods enables automatic gully feature mapping. But still few studies have specifically focused on gully feature mapping on different scales. In this study, an object-based approach to two-level gully feature mapping, including gully-affected areas and bank gullies, was developed and tested on 1-m DEM and Worldview-3 imagery of a catchment in the Chinese Loess Plateau. The methodology includes a sequence of data preparation, image segmentation, metric calculation, and random forest based classification. The results of the two-level mapping were based on a random forest model after investigating the effects of feature selection and class-imbalance problem. Results show that the segmentation strategy adopted in this paper which considers the topographic information and optimal parameter combination can improve the segmentation results. The distribution of the gully-affected area is closely related to topographic information, however, the spectral features are more dominant for bank gully mapping. The highest overall accuracy of the gully-affected area mapping was 93.06% with four topographic features. The highest overall accuracy of bank gully mapping is 78.5% when all features are adopted. The proposed approach is a creditable option for hierarchical mapping of gully feature information, which is suitable for the application in hily Loess Plateau region.

Dimensional Description of On-line Wear Debris Images for Wear Characterization

As one of the most wear monitoring indicator, dimensional feature of individual particles has been studied mostly focusing on off-line analytical ferrograph. Recent development in on-line wear monitoring with wear debris images shows that merely wear debris concentration has been extracted from on-line ferrograph images. It remains a bottleneck of obtaining the dimension of on-line particles due to the low resolution, high contamination and particle’s chain pattern of an on-line image sample. In this work, statistical dimension of wear debris in on-line ferrograph images is investigated. A two-step procedure is proposed as follows. First, an on-line ferrograph image is decomposed into four component images with different frequencies. By doing this, the size of each component image is reduced by one fourth, which will increase the efficiency of subsequent processing. The low-frequency image is used for extracting the area of wear debris, and the high-frequency image is adopted for extracting contour. Second, a statistical equivalent circle dimension is constructed by equaling the overall wear debris in the image into equivalent circles referring to the extracted total area and premeter of overall wear debris. The equivalent circle dimension, reflecting the statistical dimension of larger wear debris in an on-line image, is verified by manual measurement. Consequently, two preliminary applications are carried out in gasoline engine bench tests of durability and running-in. Evidently, the equivalent circle dimension, together with the previously developed concentration index, index of particle coverage area （IPCA）, show good performances in characterizing engine wear conditions. The proposed dimensional indicator provides a new statistical feature of on-line wear particles for on-line wear monitoring. The new dimensional feature conveys profound information about wear severity.

Local Data Analysis for Eliminating End Restraint of Triaxial Specimen

A data processing method was proposed for eliminating the end restraint in triaxial tests of soil. A digital image processing method was used to calculate the local deformations and local stresses for any region on the surface of triaxial soil specimens. The principle and implementation of this digital image processing method were introduced as well as the calculation method for local mechanical properties of soil specimens. Comparisons were made between the test results calculated by the data from both the entire specimen and local regions, and it was found that the deformations were more uniform in the middle region compared with the entire specimen. In order to quantify the nonuniform characteristic of deformation, the non-uniformity coefficients of strain were defined and calculated. Traditional and end-lubricated triaxial tests were conducted under the same condition to investigate the effects of using local region data for deformation calculation on eliminating the end restraint of specimens. After the statistical analysis of all test results, it was concluded that for the tested soil specimen with the size of 39.1 mm × 80 ram, the utilization of the middle 35 mm region of traditional specimens in data processing had a better effect on eliminating end restraint compared with end lubrication. Furthermore, the local data analysis in this paper was validated through the comparisons with the test results from other researchers.