Despite the prognostic importance of mitotic count as one of the components of the Bloom &#x2013; Richardson grade &#x5B;3&#x5D;, several studies (&#x5B;2, 9, 10&#x5D;) have found that pathologists&#x2019; agreement on the mitotic grade is fairly modest. Collecting a set of more than 4,200 candidate mitotic figures, we evaluate pathologists' agreement on individual figures, and train a computerized system for mitosis detection, comparing its performance to the classifications of three pathologists. The system&#x2019;s and the pathologists&#x2019; classifications are based on evaluation of digital micrographs of hematoxylin and eosin stained breast tissue. On figures where the majority of pathologists agree on a classification, we compare the performance of the trained system to that of the individual pathologists. We find that the level of agreement of the pathologists ranges from slight to moderate, with strong biases, and that the system performs competitively in rating the ground truth set. This study is a step towards automatic mitosis count to accelerate a pathologist's work and improve reproducibility.

Christopher Malon

Elena Brachtel

Eric Cosatto

Hans Peter Graf

Atsushi Kurata

Masahiko Kuroda

John S. Meyer

Akira Saito

Shulin Wu

Yukako Yagi

Mitotic Figure Recognition: Agreement among Pathologists and Computerized Detector

The objective is to find a practical balance between quality and performance for daily high volume whole slide imaging. We evaluated whole slide images created by various scanners at different compression factors to determine the best suitable quality factor (QF) needed for pathological images of special stains.Method: We scanned two sets of eight special stains slides each at 0.50 &#x3bc;m/pixel resolution in Hamamatsu scanner at six and five QF levels respectively to generate 72 images which were observed at a calibrated monitor by imaging specialists, a histo-technician, and a pathologist to find the most suitable QF level for special stains in digital slides.Results: Most special stains images were acceptable at QF 30 except for the stain Reticulin where the lowest acceptable QF was 50. The compression of images from QF 90 to QF 50 reduced the size of the images by 62.73&#x25;.Conclusion: 0.50 &#x3bc;m/pixel images at QF 50 or above were found suitable 12 special stain.

Anurag Sharma

Pinky Bautista

Balancing Image Quality and Compression Factor for Special Stains Whole Slide Images

In the field of whole slide imaging, the imaging device or staining process cause color variations for each slide that affect the result of image analysis made by pathologist. In order to stabilize the analysis, we developed a color standardization method and system as described below: 1) Color standardization method based on RGB imaging and multi spectral sensing, which utilize less band (16 bands) than conventional method (60 bands), 2) High speed spectral sensing module. As a result, we confirmed the following effect: 1) We confirmed the performance improvement of nucleus detection by the color standardization. And we can conduct without training data set which is needed in conventional method, 2) We can get detection performance of H&#x26;E component equivalent to conventional method (60 bands). And measurement process is more than 255 times faster.

Shinsuke Tani

Yasuhiro Fukunaga

Saori Shimizu

Munenori Fukunishi

Kensuke Ishii

Kosei Tamiya

Color Standardization Method and System for Whole Slide Imaging Based on Spectral Sensing

Pathology has recently entered the era of personalized medicine. This brings new expectations for the accuracy and precision of tissue-based diagnosis, in particular, when quantification of histologic features and biomarker expression is required. While for many years traditional pathologic diagnosis has been regarded as ground truth, this concept is no longer sufficient in contemporary tissue-based biomarker research and clinical use. Another major change in pathology is brought by the advancement of virtual microscopy technology enabling digitization of microscopy slides and presenting new opportunities for digital image analysis. Computerized vision provides an immediate benefit of increased capacity (automation) and precision (reproducibility), but not necessarily the accuracy of the analysis. To achieve the benefit of accuracy, pathologists will have to assume an obligation of validation and quality assurance of the image analysis algorithms. Reference values are needed to measure and control the accuracy. Although pathologists' consensus values are commonly used to validate these tools, we argue that the ground truth can be best achieved by stereology methods, estimating the same variable as an algorithm is intended to do. Proper adoption of the new technology will require a new quantitative mentality in pathology. In order to see a complete and sharp picture of a disease, pathologists will need to learn to use both their analogue and digital eyes.

Arvydas Laurinavicius

Aida Laurinaviciene

Darius Dasevicius

Nicolas Elie

Beno&#xee;t Plancoulaine

Catherine Bor

Paulette Herlin

Digital Image Analysis in Pathology: Benefits and Obligation

Background: Histologic image analysis plays an important role in cancer diagnosis. It describes the structure of the body tissues and abnormal structure gives the suspicion of the cancer or some other diseases. Observing the structural changes of these chaotic textures from the human eye is challenging process. However, the challenge can be defeat by forming mathematical descriptor to represent the histologic texture and classify the structural changes via a sophisticated computational method.Objective: In this paper, we propose a texture descriptor to observe the histologic texture into highly discriminative feature space.Methods: Fractal dimension describes the self-similar structures in different and more accurate manner than topological dimension. Further, the fractal phenomenon has been extended to natural structures (images) as multifractal dimension. We exploited the multifractal analysis to represent the histologic texture, which derive more discriminative feature space for classification.Results: We utilized a set of histologic images (belongs to liver and prostate specimens) to assess the discriminative power of the multifractal features. The experiment was organized to classify the given histologic texture as cancer and non-cancer. The results show the discrimination capability of multifractal features by achieving approximately 95&#x25; of correct classification rate.Conclusion: Multifractal features are more effective to describe the histologic texture. The proposed feature descriptor showed high classification rate for both liver and prostate data sample datasets.

Chamidu Atupelage

Hiroshi Nagahashi

Masahiro Yamaguchi

Michiie Sakamoto

Akinori Hashiguchi

Multifractal Feature Descriptor for Histopathology

Background: Telepathology, the practice of pathology at a long distance, has advanced continuously since 1986. The progress of telepathology passed through four stages: Static, Dynamic, Hybrid &#x26; Whole Slide Imaging.Materials and Methods: A pilot project between the Italian Hospital in Cairo &#x26; the Civico Hospital in Palermo was completed successfully, applying the static &#x26; dynamic techniques of telepathology. This project began in 2003 and continued till now. In 2004, centers in Venice, London and Pittsburgh participated actively in our project.Results: Over eight years we consulted on many problematic pathological cases with specialized pathological centers in Italy, UK &#x26; USA. In addition to the highly specialized scientific value, we saved a lot of time and money.Conclusion: We concluded from our experience that telepathology is a very useful and applicable tool for additional consulting on difficult pathological cases especially for emerging countries. In view of this success we have already established our Digital Telepathology Unit in Cairo University, using the WSI technique in teaching which was greatly successful and encouraged us to build a huge digital pathology library which will expand our telepathology &#x26; E-learning programs to cover staff and students in Egypt and Eastern Mediterranean.

Essam Ayad

Virtual Microscopy Beyond the Pyramids, Applications of WSI in Cairo University for E-Education &#x26; Telepathology

Background: Three-dimensional (3D)-reconstruction from paraffin embedded sections has been considered laborious and time-consuming. However, the high-resolution images of large object areas and different fields of view obtained by 3D-reconstruction make one wonder whether it can add a new insight into lung adenocarcinoma, the most frequent histology type of lung cancer characterized by its morphological heterogeneity.Objective: In this work, we tested whether an automated tissue sectioning machine and slide scanning system could generate precise 3D-reconstruction of microanatomy of the lung and help us better understand and define histologic subtypes of lung adenocarcinoma.Methods: Four formalin-fixed human lung adenocarcinoma resections were studied. Paraffin embedded tissues were sectioned with Kurabo-Automated tissue sectioning machine and serial sections were automatically stained and scanned with a Whole Slide Imaging system. The resulting stacks of images were 3D reconstructed by Pannoramic Viewer software.Results: Two of the four specimens contained islands of tumor cells detached in alveolar spaces that had not been described in any of the existing adenocarcinoma classifications. 3D-reconstruction revealed the details of spatial distribution and structural interaction of the tumor that could hardly be observed by 2D light microscopy studies. The islands of tumor cells extended into a deeper aspect of the tissue, and were interconnected with each other and with the main tumor with a solid pattern that was surrounded by the islands. The finding raises the question whether the islands of tumor cells should be classified into a solid pattern in the current classification.Conclusion: The combination of new technologies enabled us to build an effective 3D-reconstruction of resected lung adenocarcinomas. 3D-reconstruction may help us refine the classification of lung adenocarcinoma by adding detailed spatial/structural information to 2D light microscopy evaluation.

Maristela L. Onozato

Veronica E. Klepeis

Mari Mino-Kenudson

A Role of Three-Dimensional (3D)-Reconstruction in the Classification of Lung Adenocarcinoma

Background: Making an automatic diagnosis based on virtual slides and whole slide imaging or even determining whether a case belongs to a single class, representing a specific disease, is a big challenge. In this work we focus on WHO Classification of Tumours of the Central Nervous System. We try to design a method which allows to automatically distinguish virtual slides which contain histopathologic patterns characteristic of glioblastoma &#x2013; pseudopalisading necrosis and discriminate cases with neurinoma (schwannoma), which contain similar structures &#x2013; palisading (Verocay bodies).Methods: Our method is based on computer vision approaches like structural analysis and shape descriptors. We start with image segmentation in a virtual slide, find specific patterns and use a set of features which can describe pseudopalisading necrosis and distinguish it from palisades. Type of structures found in a slide decides about its classification.Results: Described method is tested on a set of 49 virtual slides, captured using robotic microscope. Results show that 82&#x25; of glioblastoma cases and 90&#x25; of neurinoma cases were correctly identified by the proposed algorithm.Conclusion: Our method is a promising approach to automatic detection of nervous system tumors using virtual slides.

Slawomir Walkowski

Janusz Szymas

Histopathologic Patterns of Nervous System Tumors Based on Computer Vision Methods and Whole Slide Imaging (WSI)

Background: Intraoperative consultations in neuropathology are often assessed by smear preparations rather than by frozen sections. Both techniques are standard practice for light microscopic examination on site, but there is little data comparing these techniques in a telepathology setting.Methods: Thirty cases of brain tumours submitted for intraoperative consultation at our institution between July and December 2010 were identified in which both frozen section and tissue smear preparations were available for digitization at 20&#xd7; magnification. Slides were digitized using a Hamamatsu Nanozoomer 2.0 HT whole slide scanner, and resulting digital images were visualized at 1680 &#xd7; 1050 pixel resolution with NDP. view software.Results: The original intraoperative diagnosis was concordant with the sign out diagnosis in 29/30 cases; one tumeur was initially interpreted as a high grade glioma but proved to be a lymphoma at sign out.Digitized frozen section slides were sufficient for diagnosis at 10&#xd7; magnification in 27/30 cases. Digitized tissue smears were sufficient for diagnosis at 10&#xd7; magnification in 28/30 cases. In two cases tumour was present on the tissue smear but not the frozen section (one case of recurrent astrocytoma, one case of meningeal carcinomatosis). In one case of lymphoma, tumour was present on frozen section only. These discrepancies were attributed to tissue sampling rather than image quality.Examination of digitized slides at higher magnfication (20&#xd7;) permitted confirmation of mitoses and Rosenthal fibers on tissue smear preparations, but did not change the primary diagnosis. Intra-slide variations in tissue thickness on smear preparations led to variable loss of focus in digitized images, but did not affect image quality in thinner areas of the smear or impede diagnosis.Conclusion: Digitized tissue smears are suitable for intraoperative neurotelepathology and provide comparable information to digitized frozen sections at medium power magnification.

Peter V. Gould

Stephan Saikali

A Comparison of Digitized Frozen Section and Smear Preparations for Intraoperative Neurotelepathology

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Analytical Cellular Pathology provides a forum for pathologists and medical practitioners working in the cellular pathology field. Topics covered include cytology, carcinogenesis, cell receptors, biomarkers, diagnostic pathology, and immunopathology.

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Chief Editor&nbsp;Professor Dimitrios Karamichos&nbsp;focuses on investigating corneal wound healing and dystrophies with a particular interest in the effect of transforming growth factor-β3 or TGF- β3 on corneal stromal cells and their extracellular environment.

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