Fibrosis distribution in explanted cirrhotic livers

Hall A, Germani G, Isgrò G, Burroughs A K & Dhillon A P
(2012) Histopathology  60, 270–277 Fibrosis distribution in explanted cirrhotic livers Aims: Little information is available regarding the distribution of fibrosis within cirrhotic livers. We measured collagen in cirrhotic explants to determine if fibrosis differs (i) between left (L) and right (R) lobes, and (ii) between different aetiologies. Methods and results: Ten cases each of common aetiologies of cirrhosis were studied: alcoholic liver disease (ALD), primary biliary cirrhosis (PBC), primary sclerosing cholangitis (PSC), autoimmune hepatitis (AIH), hepatitis C virus (HCV) and hepatitis B virus (HBV). A total of 120 tissue blocks (one block each from L and R lobes) were studied. Collagen was measured as collagen proportionate area (CPA), i.e. the proportion of the tissue sections stained by picro‐Sirius red. L and R lobes contained similar amounts of fibrosis (r = 0.788; P < 0.0001) with good agreement between L and R lobes (Bland–Altman analysis, R lobe bias = 1.35%). Median CPA across all aetiologies (R plus L lobes) was 21.5%, (L = 8–40%, R = 10–47%). There was more fibrosis in ALD (30%, 15–47%) than PBC (23.5%, 16–34%) and PSC (22.5%, 8–33%), which in turn showed more than AIH (18.5%, 10–40%), HCV (17%, 13–31%) and HBV (16.5%, 8–30%). Conclusions: At the time of transplantation cirrhotic livers have different ranges of collagen proportionate area, according to aetiology. R lobe fibrosis corresponds with L lobe fibrosis. The range of fibrosis within each aetiological group could be useful for prognostic subclassification.     

医用显微图像的计算机处理及形态量化分析

本文介绍了自行研制开发出的一套以测量分析医用显微图像的形态参数为主要特征的图像处理软件系统。文中以新生大鼠缺氧缺血脑切处中线粒体衰竭以及大鼠颈动脉损伤后内膜增生抑制作用的形态参数的测量为例,证实本软件可广泛应用于临床诊断,治疗等研究中。     

Type IV collagen immunostaining and computerized image analysis (SAMBA) in breast and endometrial disorders

Type IV collagen immunostaining was performed on tissue sections from a large series of non-malignant and malignant disorders of the breast and endometrium. The results were analysed by means of a computerized system of image analysis referred to as SAMBA. It was shown that this system provided an accurate, reliable, reproducible, automated and multiparameteric analysis of collagen IV immunoprecipitates. It was concluded that this standardized method of analyses can be routinely used for the measurement of collagen IV, thus enabling correlations to be sought with histopathological and clinical data.     

Immunolocalization of type III collagen and procollagen in cirrhotic human liver using monoclonal antibodies

Summary Immunolocalization of type III collagen and procollagen in cirrhotic human liver was studied using monoclonal antibody specific for the helical determinant of type III collagen extracted from human placenta. Deparaffinized, trypsin-treated cirrhotic liver sections from 8 autopsy cases were examined by the unlabeled peroxidase-antiperoxidase and immunofluorescence techniques. These techniques revealed the localization of this epitope shared by type III collagen and procollagen not only in the extracellular matrix of hepatocytes and sinusoidal cells but also in the cytoplasm. In hepatocellular carcinoma concurrent with cirrhosis, neoplastic cells were shown to react with this antibody as well.These results are consistent with data obtained using antiserum specific for bovine type III procollagen aminopeptide which appeared in our previous report.     

Microvessel area using automated image analysis is reproducible and is associated with prognosis in breast cancer

Microvessel density may be one measure of tumor associated angiogenesis but is methodologically difficult to standardize and reproduce. We used our automated quantitative image analysis system, AQUA, to more objectively assess microvessel area. Cytokeratin and CD31 were used to create tumor and vessel compartments respectively with AQUA. Microvessel area was defined as CD31 compartment area normalized to the tissue spot area (CD31 area/area of entire tissue spot). Consecutive breast cancer whole sections were stained with CD31 to compare pathologist-based microvessel density with AQUA microvessel area. Microvessel areas of 3-fold redundant tissue microarrays of 652 primary breast cancers were also assessed. CD34 and factor VIII-related antigen were also tested. There was nearly linear correlation between pathologist's microvessel density and AQUA microvessel area with regression coefficient R = 0.846. On the redundant arrays, of the 67% evaluable cases, 52% were microvessel area high and 48% low with good reproducibility of scores (Spearman rho 0.551). AQUA microvessel area was associated with larger tumors, node positivity, and estrogen receptor negativity, with 20 year survival at the univariate and multivariate levels (P < .0001 and P = .0121, respectively). CD34 or factor VIII-related antigen were more heterogenous, had poor association with CD31, and did not correlate with outcome. AQUA-based microvessel area was significantly correlated with both standard breast cancer prognostic parameters as well as with clinical outcome. In the future, it may also allow the use of the AQUA-based algorithms to quantify the expression of angiogenic biomarkers to either tumor or microvessel area-specific compartments.     

Diagnosis of chronic atrophic gastritis by morphometric image analysis. A new method to overcome the confounding effect of the inflammatory infiltrate

The risk of gastric cancer increases with the severity of gastric mucosal atrophy. Atrophy is a 'loss of properly specialized glands'. These glands may be substituted by metaplastic cells and by interstitial fibrosis, or displaced by an inflammatory infiltrate. Agreement among pathologists for the diagnosis of atrophy is poor (kappacoefficient < 0.4), probably because inflammatory infiltrate can confound the identification of gland loss. The aim of this study was to evaluate interstitial fibrosis by image analysis, and thereby overcoming the confounding effect of the inflammatory infiltrate. Gastric biopsies of 40 controls (20 children and 20 adults) and 111 patients with chronic atrophic gastritis were examined. Patients underwent another biopsy a year later. Gastric sections were examined by conventional histology (updated Sydney system) and image analysis to detect collagen and non-collagen fibres. There were no significant intra- or inter-operator differences in the evaluation by image analysis of fibre content in either controls or patients. In both controls and patients, the mean percentage of collagen fibres was lower in the gastric body (9%) than in the antrum (10%). In the antrum it was 14%, 17% and 20% in patients with mild, moderate and severe atrophy, respectively. A year later, histology showed that the grade of atrophy had decreased in 42%, probably due to the regression of inflammation, and increased in 10% of cases, but interstitial fibrosis (expressed as collagen fibre content) was practically unchanged. The use of image analysis of gastric biopsies appears to be a reliable method with which to measure interstitial fibrosis, even in the presence of an inflammatory infiltrate. This study highlights the difference between 'real gastric atrophy', where glands are replaced by collagen fibres, and 'apparent gastric atrophy', where glands are displaced by an inflammatory infiltrate.     

Quantitative assessment of basement membranes in soft tissue tumours. Computerized image analysis of laminin and type IV collagen

Basement membranes (BMs) in 201 soft tissue tumours were quantified using computerized image analysis of tissues immunostained for laminin and type IV collagen. The purpose of the study was to compare and quantify the extent of BM deposition in a large and varied group of benign and malignant tumours. Laminin and type IV collagen gave similar results. The difference between benign and malignant was statistically highly significant (P=0·0001), with greater deposition in benign tumours. BM deposition was homogeneous in benign tumours and heterogeneous in sarcomas and appeared to correlate with the degree of differentiation. Some poorly differentiated sarcomas showed cytoplasmic laminin staining but little or no extracellular BM. Immunohistochemical evaluation of BM has some advantages over electron microscopy; specialized equipment is not needed and since large samples can be studied with little sampling error, heterogeneity can be studied more readily. Subjective visual assessment gives a good overall indication of the extent of BM deposition and in many situations is likely to be a suitable alternative to image analysis. Because of staining heterogeneity, BM immunohistochemistry is unlikely to be of significant value in the diagnosis of specific types of sarcoma. © 1998 John Wiley & Sons, Ltd.     

影响DR图像质量因素分析

目的研究影响DR图像质量因素和规避。方法通过DR成像系统成像后,经工作站后处理,调整至最佳传至激光相机打片。选取600幅照片,其中男性326例,女性274例;年龄2天～78岁,平均年龄45岁。照片所摄部位以胸片、脊柱片、四肢骨和关节片为主,通过X射线评片标准,对其质量进行评价,探讨影响成像质量因素。结果甲级片498张,非甲级片102张。通过着重对非甲级片分析,影响DR图像质量因素如下。①技术人员的操作技能和业务技术水平。此原因所致68张,占66%。②摄片技术参数。此原因所致23张,占23%。③机器性能。此原因所致3张,占3%。④图像后处理技术。此原因所致6张,占6%。⑤激光相机。此原因所致2张,占2%。结论影响DR照片质量因素很多,技术人员的操作技能和水平是其主要因素。应用DR设备过程中,只有不断提高技术人员的综合素质水平,规范操作和不断总结使用经验,增强对设备原理的学习,探索更多临床经验,才能得到质量更佳图像照片。     

Quantitative morphology of human cirrhotic livers. Part I: Parameter expression of pattern as a basis for computerized classification

Although classification is a long-used method of histopathology, a reproducible one has yet to be created. We established a most adequate classification of cirrhosis from a geometric and statistical point of view, by reducing its form to a set of quantities and submitting the data to multivariate analysis. In this article, methods for quantification are described as a preliminary step for the Statistical treatment that appears in another paper. The pattern was reduced to a set of four quantities: (i) the mean nodular radius: (ii) the coarseness; (iii) the mean septal thickness; and (iv) the degree of nodular separation. A model of dispersed spheres with various radii r was employed to assimilate cirrhosis; r was assumed to follow a logarithmic normal distribution. The parameters of this distribution were estimated stereologically from measurements on microscopic sections of chord lengths Λ generated from nodules by a test line. The coarseness was defined as the volume % of nodules larger than 1.5 mm in r . The mean septai thickness was determined stereologically on a plate model, into which the actual septa were transformed without changing their volume or surface density. The degree of nodular separation p_θ was defined as a two-dimensional parameter, based on the curvature of nodulo-septal borders. It was demonstrated in several examples how accurately a set of these quantltles describes various patterns of cirrhosis.     

Quantification of collagen and elastic fibers using whole‐slide images of liver biopsy specimens

Histological evaluation of fibrosis after a liver biopsy is crucial for evaluating the pathology of patients with chronic liver disease. Previous studies have reported quantitative analyses of fibrosis using images of collagen‐stained sections. However, analysis of these studies requires manual selection of the region of interest. In addition, the quantification of elastic fibers is not considered. The present study was conducted in order to measure both the collagen and elastic fiber area ratios using Elastica van Gieson‐stained whole‐slide images (WSIs) of liver biopsy specimens. High‐resolution WSIs provide precise color classification, enabling accurate detection of even fine collagen and elastic fibers. To minimize the influence of pre‐existing fibrous tissue, median area ratios of the collagen and elastic fibers were independently calculated from the image tiles of the WSIs. These median area ratios were highly concordant with area ratios after the pre‐existing fibrous tissues were manually trimmed from the WSI. Further, these median area ratios were correlated with liver stiffness as measured by transient elastography (collagen: r = 0.73 [P < 0.01], elastic: r = 0.53 [P < 0.01]). Our approach to quantifying liver fibrosis will serve as an effective tool to evaluate liver diseases in routine practice.     

眼底数码图像血管分割预处理关键技术研究

目的本文以眼底图像为研究对象,针对眼底图像血管分割的预处理关键技术进行了深入的研究和探讨。方法首先通过RGB通道选择,显示出较为清晰的血管图像,并通过自适应直方图均衡化增强图像对比度。其次,利用灰度倒置和灰度增强,使细小血管更加清晰,并采用空域滤波对血管轮廓边缘进行增强。最后进行图像填充,去除背景中的噪声干扰。结果通过MATLAB仿真实验,获得了血管脉络清晰的眼底数码图像。结论本研究为血管的自动分割以及血管成分的定性和定量分析奠定了基础。     

The use of digital images in pathology

Digital images are routinely used by the publishing industry, but most diagnostic pathologists are unfamiliar with the technology and its possibilities. This review aims to explain the basic principles of digital image acquisition, storage, manipulation and use, and the possibilities provided not only in research, but also in teaching and in routine diagnostic pathology. Images of natural objects are usually expressed digitally as ‘bitmaps’—rectilinear arrays of small dots. The size of each dot can vary, but so can its information content in terms, for example, of colour, greyscale or opacity. Various file formats and compression algorithms are available. Video cameras connected to microscopes are familiar to most pathologists; video images can be converted directly to a digital form by a suitably equipped computer. Digital cameras and scanners are alternative acquisition tools of relevance to pathologists. Once acquired, a digital image can easily be subjected to the digital equivalent of any conventional darkroom manipulation and modern software allows much more flexibility, to such an extent that a new tool for scientific fraud has been created. For research, image enhancement and analysis is an increasingly powerful and affordable tool. Morphometric measurements are, after many predictions, at last beginning to be part of the toolkit of the diagnostic pathologist. In teaching, the potential to create dramatic yet informative presentations is demonstrated daily by the publishing industry; such methods are readily applicable to the classroom. The combination of digital images and the Internet raises many possibilities; for example, instead of seeking one expert diagnostic opinion, one could simultaneously seek the opinion of many, all around the globe. It is inevitable that in the coming years the use of digital images will spread from the laboratory to the medical curriculum and to the whole of diagnostic pathology. © 1997 John Wiley & Sons, Ltd.     

Kinematic analysis of human walking gait using digital image processing

A system using digital image processing techniques for kinematic analysis of human gait has been developed. The system is cheap, easy to use, automated and provides useful detailed quantitative information to the medical profession. Passive markers comprising black annuli on white card are placed on the anatomical landmarks of the subject. Digital images at the standard television rate of 25 per second are acquired of the subject walking past a white background. The images are obtained, stored and processed using standard commercially available hardware, i.e. video camera, video recorder, digital framestore and an IBM PC. Using a single-threshold grey level, all the images are thresholded to produce binary images. An automatic routine then uses a set of pattern recognition algorithms to locate accurately and consistently the markers in each image. The positions of the markers are analysed to determine to which anatomical landmark they correspond, and thus a stick diagram for each image is obtained. There is also a facility where the positions of the markers may be entered manually and errors corrected. The results may be presented in a variety of ways: stick diagram animation, sagittal displacement graphs, flexion diagrams and gait parameters.     

Parotid gland tumors: Correlation between routine cytology and cytomorphometry by digital image analysis using conventional and newly introduced cytomorphometric parameters

The objective of this study was to compare qualitative cytomorphology and morphometric characteristics of parotid gland tumor cells, with the aid of a computer‐assisted system of image analysis. Routine qualitative cytologic and quantitative morphometric results from 64 parotid gland tumors were compared. Ultrasound (US)‐guided fine‐needle aspiration (FNA) specimens were taken from 54 patients. Eleven conventionally used morphometric parameters were studied: area, perimeter, convex area, convexity, maximal and minimal radius, length, breadth, form factor (FF), elongation factor, and nuclear‐ cytoplasmatic (N/C) ratio. Two newly introduced nuclear form factors were also measured: area symmetry factor and perimeter symmetry factor. The following nuclear morphometric parameters were significantly different between malignant and benign tumors: area, perimeter, convex area, convexity, maximal and minimal radius, length, breadth, FF, elongation factor, area symmetry factor, and perimeter symmetry factor. Comparing the cutoff values and receiver operating characteristic (ROC) curves the following nuclear morphometric parameters were found most useful in separating benign from malignant tumors: area, perimeter, convex area, maximal radius, length, and FF. The following whole cell morphometric parameters were significantly different between malignant and benign tumors: minimal and maximal radius, convexity, breadth, FF, and elongation factor. N/C ratio was significantly higher in malignant tumors. The quantitative morphometric analysis is a useful tool in the cytological differentiation between benign and malignant parotid gland tumors. Computerized image analysis may add to morphological evaluation by turning qualitative data into quantitative values. Diagn. Cytopathol. 2013;41:776–784. © 2013 Wiley Periodicals, Inc.