When using different cameras and displays in the shot and display of one subject, different color images are often showed. To solve this problem, we have developed a color chart in which the constituent colors dyed with dyes are evenly distributed in the color space. We have also developed a tool that creates an International Color Consortium (ICC) profile from the captured image of this color chart. In this report, we will describe the color difference correction accuracy when our method is applied to actual stained pathological specimens taken with multiple whole-slide imaging (WSI). We confirmed color correction accuracy of Lab values in major parts such as the cell nucleus. The Lab value of the specimen itself measured by a spectrocolorimeter was compared with that of the captured image. As a result, the color difference ΔE in H&E-stained cell nucleus was improved from 32.2 to 6.4 for Nanozoomer and from 13.5 to 7.2 for ultra-fast scanner (UFS) by our color correction. The results of the evaluations for other areas and other stain methods (PAS, EVG, MT, and PAM) were good. In the future, high-accuracy color correction of teacher data/evaluation data in AI diagnosis using pathological images will be important.
The pathological diagnosis of a transplanted kidney is made on Banff Classification in order to gain an accurate
understanding of the condition of the kidney. This type of diagnosis is extremely difficult and, thus, a variety of methods
for diagnosis, including diagnosis by electron microscope, are being considered at present. Quantification of the
diagnostic information derived by image processing is required for such purposes. This study proposes an automatic
extraction method for normal glomeruli for the purpose of quantifying Elastica Van Gieson(EVG)-stained pathology
specimens. In addition, we provide a report on the package of methods that we have created for the extraction of the
glomerulus in the cortex.
This paper proposes a digital image analysis method to support quantitative pathology by automatically segmenting the hepatocyte structure and quantifying its morphological features. To structurally analyze histopathological hepatic images, we isolate the trabeculae by extracting the sinusoids, fat droplets, and stromata. We then measure the morphological features of the extracted trabeculae, divide the image into cords, and calculate the feature values of the local cords. We propose a method of calculating the nuclear–cytoplasmic ratio, nuclear density, and number of layers using the local cords. Furthermore, we evaluate the effectiveness of the proposed method using surgical specimens. The proposed method was found to be an effective method for the quantification of the Edmondson grade.
The steatosis in liver pathological tissue images is a promising indicator of nonalcoholic fatty liver disease (NAFLD) and the possible risk of hepatocellular carcinoma (HCC). The resulting values are also important for ensuring the automatic and accurate classification of HCC images, because the existence of many fat droplets is likely to create errors in quantifying the morphological features used in the process. In this study we propose a method that can automatically detect, and exclude regions with many fat droplets by using the feature values of colors, shapes and the arrangement of cell nuclei. We implement the method and confirm that it can accurately detect fat droplets and quantify the fat droplet ratio of actual images. This investigation also clarifies the effective characteristics that contribute to accurate detection.
Diagnosis of hepatocellular carcinoma (HCC) on the basis of digital images is a challenging problem because, unlike gastrointestinal carcinoma, strong structural and morphological features are limited and sometimes absent from HCC images. In this study, we describe the classification of HCC images using statistical distributions of features obtained from image analysis of cell nuclei and hepatic trabeculae. Images of 130 hematoxylin-eosin (HE) stained histologic slides were captured at 20X by a slide scanner (Nanozoomer, Hamamatsu Photonics, Japan) and 1112 regions of interest (ROI) images were extracted for classification (551 negatives and 561 positives, including 113 well-differentiated positives). For a single nucleus, the following features were computed: area, perimeter, circularity, ellipticity, long and short axes of elliptic fit, contour complexity and gray level cooccurrence matrix (GLCM) texture features (angular second moment, contrast, homogeneity and entropy). In addition, distributions of nuclear density and hepatic trabecula thickness within an ROI were also extracted. To represent an ROI, statistical distributions (mean, standard deviation and percentiles) of these features were used. In total, 78 features were extracted for each ROI and a support vector machine (SVM) was trained to classify negative and positive ROIs. Experimental results using 5-fold cross validation show 90% sensitivity for an 87.8% specificity. The use of statistical distributions over a relatively large area makes the HCC classifier robust to occasional failures in the extraction of nuclear or hepatic trabecula features, thus providing stability to the system.
Recent advances in information technology have improved pathological virtual-slide technology and diagnostic support system studies of pathological images. Diagnostic support systems utilize quantitative indices determined by image processing. In previous studies on diagnostic support systems, carcinomatous areas of breast or lung have been
recognized by the feature quantities of nuclear sizes, complexities, and internuclear distances based on graph theory,
among other features. Improving recognition accuracy is important for the addition of new feature quantities. We
focused on hepatocellular carcinoma (HCC) and investigated new feature quantities of histological images of HCC. One of the most important histological features of HCC is the trabecular pattern. For diagnosing cancer, it is important to recognize the tumor cell trabeculae. We propose a new algorithm for calculating the number of cell layers in histological images of HCC in tissue sections stained by hematoxylin and eosin. For the calculation, we used a Delaunay diagram that was based on the median points of nuclei, deleted the sinusoid and fat droplet regions from the Delaunay diagram, and counted the Delaunay lines while applying a thinning algorithm. Moreover, we experimented with the calculation of the number of cell layers with our method for different histological grades of HCC. The number of cell layers discriminated tumor differentiations and Edmondson grades; therefore, our algorithm may serve as an index of HCC for diagnostic support systems.
The analysis of hepatic tissue structure is required for quantitative assessment of liver histology. Especially, a cord-like
structure of liver cells, called trabecura, has important information in the diagnosis of hepatocellular carcinoma (HCC).
However, the extraction of trabeculae is thought to be difficult because liver cells take on various colors and appearances depending on tissue conditions. In this paper, we propose an approach to extract trabeculae from images of hematoxyline and eosin stained liver tissue slide by extracting the rest of trabeculae: sinusoids and stromal area. The sinusoids are simply extracted based on the color information, where the image is corrected by an orientation selective filtering before segmentaion. The stromal area mainly consists of fiber, and often includes lymphocytes densely. Therefore, in the proposed method, fiber region and lymphocytes are extracted separately, then, stromal region is determined based on the extracted results. The determination of stroma is performed based on superpixels, to obtain precise boundaries. Once the regions of sinusoids and stroma are obtained, trabeculae can be segmented as the remaining region. The proposed method was applied to 10 test images of normal and HCC liver tissues, and the results were evaluated based on the manual segmentation. As a result, we confirmed that both sensitivity and specificity of the extraction of trabeculae reach around 90%.
The presence of a liver disease such as cirrhosis can be determined by examining the proliferation of
collagen fiber from a tissue slide stained with special stain such as the Masson's trichrome(MT) stain.
Collagen fiber and smooth muscle, which are both stained the same in an H&E stained slide, are stained
blue and pink respectively in an MT-stained slide. In this paper we show that with multispectral imaging
the difference between collagen fiber and smooth muscle can be visualized even from an H&E stained
image. In the method M KL bases are derived using the spectral data of those H&E stained tissue
components which can be easily differentiated from each other, i.e. nucleus, cytoplasm, red blood cells,
etc. and based on the spectral residual error of fiber weighting factors are determined to enhance spectral
features at certain wavelengths. Results of our experiment demonstrate the capability of multispectral
imaging and its advantage compared to the conventional RGB imaging systems to delineate tissue
structures with subtle colorimetric difference.
Physical staining is indispensable in pathology. While physical staining uses chemicals, "digital staining" exploits the
differing spectral characteristics of the different tissue components to simulate the effect of physical staining. Digital
staining for pathological images involves two basic processes: classification of tissue components and digital
colorization whereby the classified tissue components are impressed with colors associated to their reaction to specific
dyes. Spectral features, i.e. spectral transmittance, of the different tissue structures are dependent on the staining
condition of the tissue slide. Thus, if the staining condition of the test image is different, classification result is affected,
and the resulting digitally-stained image may not reflect the desired result. This paper shows that it is possible to obtain
robust classification results by correcting the dye amount of each test-image pixel using Beer Lambert's Law. Also the
effectiveness of such technique to be incorporated to the current digital staining scheme is investigated as well.
Staining of tissue specimens is a classical procedure in pathological diagnosis to enhance the contrast between tissue components such that identification and classification of these components can be easily performed. In this paper, a framework for digital staining of pathological specimens using the information derived from the L-band spectral transmittance of various pathological tissue components is introduced, particularly the transformation of a Hematoxylin and Eosin (HE) stained specimen to its Masson-Trichrome (MT) stained counterpart. The digital staining framework involves the classification of tissue components, which are highlighted when the specimen is actually stained with MT stain, e.g. fibrosis, from the HE-stained image; and the linear mapping between specific sets of HE and MT stained transmittance spectra through pseudo-inverse procedure to produce the LxL transformation matrices that will be used to transform the HE stained transmittance to its equivalent MT stained transmittance configuration. To generate the digitally stained image, the decisions of multiple quadratic classifiers are pooled to form the weighting factors for the transformation matrices. Initial results of our experiments on liver specimens show the viability of multispectral imaging (MSI) for the implementation of digital staining in the pathological context.
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