Analysis of Signal Decomposition and Stain Separation methods for biomedical applications

Nowadays, the biomedical signal processing and classification and medical image interpretation play an essential role in the detection and diagnosis of several human diseases. The problem of high variability and heterogeneity of information, which is extracted from digital data, can be addressed with signal decomposition and stain separation techniques which can be useful approaches to highlight hidden patterns or rhythms in biological signals and specific cellular structures in histological color images, respectively. This thesis work can be divided into two macro-sections. In the first part (Part I), a novel cascaded RNN model based on long short-term memory (LSTM) blocks is presented with the aim to classify sleep stages automatically. A general workflow based on single-channel EEG signals is developed to enhance the low performance in staging N1 sleep without reducing the performances in the other sleep stages (i.e. Wake, N2, N3 and REM). In the same context, several signal decomposition techniques and time-frequency representations are deployed for the analysis of EEG signals. All extracted features are analyzed by using a novel correlation-based timestep feature selection and finally the selected features are fed to a bidirectional RNN model. In the second part (Part II), a fully automated method named SCAN (Stain Color Adaptive Normalization) is proposed for the separation and normalization of staining in digital pathology. This normalization system allows to standardize digitally, automatically and in a few seconds, the color intensity of a tissue slide with respect to that of a target image, in order to improve the pathologist’s diagnosis and increase the accuracy of computer-assisted diagnosis (CAD) systems. Multiscale evaluation and multi-tissue comparison are performed for assessing the robustness of the proposed method. In addition, a stain normalization based on a novel mathematical technique, named ICD (Inverse Color Deconvolution) is developed for immunohistochemical (IHC) staining in histopathological images. In conclusion, the proposed techniques achieve satisfactory results compared to state-of-the-art methods in the same research field. The workflow proposed in this thesis work and the developed algorithms can be employed for the analysis and interpretation of other biomedical signals and for digital medical image analysis

Bayesian K-SVD for H and E blind color deconvolution. Applications to stain normalization, data augmentation and cancer classification

This work was supported by project PID2019-105142RB-C22 funded by MCIN / AEI / 10.13039 / 501100011033, Spain, and project P20_00286 funded by FEDER /Junta de Andalucía-Consejería de Transformación Económica, Industria, Conocimiento y Universidades, Spain. The work by Fernando Pérez-Bueno was sponsored by Ministerio de Economía, Industria y Competitividad , Spain, under FPI contract BES-2017-081584 . Funding for open access charge: Universidad de Granada / CBUA, Spain.Stain variation between images is a main issue in the analysis of histological images. These color variations, produced by different staining protocols and scanners in each laboratory, hamper the performance of computer-aided diagnosis (CAD) systems that are usually unable to generalize to unseen color distributions. Blind color deconvolution techniques separate multi-stained images into single stained bands that can then be used to reduce the generalization error of CAD systems through stain color normalization and/or stain color augmentation. In this work, we present a Bayesian modeling and inference blind color deconvolution framework based on the K-Singular Value Decomposition algorithm. Two possible inference procedures, variational and empirical Bayes are presented. Both provide the automatic estimation of the stain color matrix, stain concentrations and all model parameters. The proposed framework is tested on stain separation, image normalization, stain color augmentation, and classification problems.CBUAJunta de Andalucía-Consejería de Transformación Económica, Industria, Conocimiento y UniversidadesFamily Process Institute BES-2017-081584Universidad de GranadaEuropean Regional Development FundMinisterio de Economía, Industria y Competitividad, Gobierno de EspañaAgencia Estatal de Investigación P20_0028

A hybrid approach for stain normalisation in digital histopathological images

Stain in-homogeneity adversely affects segmentation and quantifi-cation of tissues in histology images. Stain normalisation techniques have been used to standardise the appearance of images. However, most the available stain normalisation techniques only work on a particular kind of stain images. In addition, some of these techniques fail to utilise both the spatial and tex-tural information in histology images, leading to image tissue distortion. In this paper, a hybrid approach has been developed, based on an octree colour quantisation algorithm combined with the Beer-Lambert law, a modified blind source separation algorithm, and a modified colour transfer approach. The hybrid method consists of two stages the stain separation stage and colour transfer stage. An octree colour quantisation algorithm combined with Beer-Lambert law, and a modified blind source separation algorithm are used during the stain separation stage to computationally estimate the amount of stain in an histology image based on its chromatic and luminous response. A modified colour transfer algorithm is used during the colour transfer stage to minimise the effect of varying staining and illumination. The hybrid method addresses the colour variation problem in both H&DAB (Haemotoxylin and Diaminoben-zidine) and H&E (Haemotoxylin and Eosin) stain images. The stain normali-sation method is validated against ground truth data. It is widely known that the Beer-Lambert law applies to only stains (such as haematoxylin, eosin) that absorb light. We demonstrate that the Beer-Lambert law applies is applicable to images containing a DAB stain. Better stain normalisation results are obtained in both H&E and H&DAB images

Variational Bayesian Blind Color Deconvolution of Histopathological Images

2019 IEEE. Personal use is permitted, but republication/redistribution requires IEEE permission.Most whole-slide histological images are stained with two or more chemical dyes. Slide stain separation or color deconvolution is a crucial step within the digital pathology workflow. In this paper, the blind color deconvolution problem is formulated within the Bayesian framework. Starting from a multi-stained histological image, our model takes into account both spatial relations among the concentration image pixels and similarity between a given reference color-vector matrix and the estimated one. Using Variational Bayes inference, three efficient new blind color deconvolution methods are proposed which provide automated procedures to estimate all the model parameters in the problem. A comparison with classical and current state-of-the-art color deconvolution algorithms using real images has been carried out demonstrating the superiority of the proposed approach.Visual Information Processing (Ref. TIC-116

Automated characterization of Tumor-Infiltrating Lymphocytes (TIL) in histological breast images

Cancer illness has a big influence on society. Its extended proliferation and high aggressiveness make it a difficult problem to solve and therefore a big deal for science. Recently, a research trend has been focusing on how 3D tumor structure affects the development of the cancer and its outcome, especially metastasis. Stromal structure and tumor cell signaling are processes that highly influence tumor migration. Thus, histological analysis becomes a fundamental tool to study tumor structure, which provides valuable information about cell characteristics and organization. The relevance of histological study is supported by the increasing interest of anatomopathologists to have good automatic solutions to support the specialist’s diagnosis. For this purpose, the current thesis proposes an automated approach to analyze hematoxylin and eosin (H&E) stained histological images, particularly coming from breast cancer patients. The proposed method consists on the classification of the nuclei in H&E-stained histological images and the further analysis of tumor-infiltrating lymphocytes (TIL) present on the visualized section. The starting point of the approach is the automatic nuclei-segmented binary mask. Each of the segmented nuclei is classified into two types, cancerous or healthy. The classification is performed by a trained artificial neural network to give two binary masks, each of them containing one type of nuclei. Then, the algorithm can follow two different paths: classification of zones or TIL analysis. Classification of zones has the aim to provide a more comfortable support to perform cancer diagnosis, because it provides quantitative information of tumor lobule size. To achieve it, a nuclei correction step is executed, by which each nucleus class depends on the area surrounding it. In this way, a clearer vision of the existing zones is provided (tumor lobule or tumor microenvironment). The other approach is to perform TIL analysis. This technique is based on the nuclei classified binary masks and analyzes the immune system response against the tumor. This way, healthy cells of tumor microenvironment are detected and quantified. The ratio of TIL occupied area to free microenvironment area is computed as informational parameter. This ratio is calculated by the combination of a manually-segmented zone binary mask and the nuclei classified binary mask. In this way, only healthy nuclei of microenvironment zone are considered, dividing the sum of their area by the free sections of the microenvironment zone (i.e. area of microenvironment zone where nuclei are not present). Moreover, the TIL dispersion factor is computed to study their distribution throughout the area by dividing the microenvironment area in several zones and calculate the standard deviation of the area of lymphocytes within each of them. Afterward, the opposed of standard deviation is computed to obtain the dispersion factor. Automatic results are found to match the gold standard (the pathologist’s diagnosis), although some error is observed after evaluation. The approach taken in this work has a positive outlook, even though some aspects need to be polished, like the algorithm accuracy and the use of a larger set of images to claim a proper functionality for global cases.Ingeniería Biomédic

RECENT CNN-BASED TECHNIQUES FOR BREAST CANCER HISTOLOGY IMAGE CLASSIFICATION

Histology images are extensively used by pathologists to assess abnormalities and detect malignancy in breast tissues. On the other hand, Convolutional Neural Networks (CNN) are by far, the privileged models for image classification and interpretation. Based on these two facts, we surveyed the recent CNN-based methods for breast cancer histology image analysis. The survey focuses on two major issues usually faced by CNN-based methods namely the design of an appropriate CNN architecture and the lack of a sufficient labelled dataset for training the model. Regarding the design of the CNN architecture, methods examining breast histology images adopt three main approaches: Designing manually from scratch the CNN architecture, using pre-trained models and adopting an automatic architecture design. Methods addressing the lack of labelled datasets are grouped into four categories: methods using pre-trained models, methods using data augmentation, methods adopting weakly supervised learning and those adopting feedforward filter learning. Research works from each category and reported performance are presented in this paper. We conclude the paper by indicating some future research directions related to the analysis of histology images

Differently stained whole slide image registration technique with landmark validation

Abstract. One of the most significant features in digital pathology is to compare and fuse successive differently stained tissue sections, also called slides, visually. Doing so, aligning different images to a common frame, ground truth, is required. Current sample scanning tools enable to create images full of informative layers of digitalized tissues, stored with a high resolution into whole slide images. However, there are a limited amount of automatic alignment tools handling large images precisely in acceptable processing time. The idea of this study is to propose a deep learning solution for histopathology image registration. The main focus is on the understanding of landmark validation and the impact of stain augmentation on differently stained histopathology images. Also, the developed registration method is compared with the state-of-the-art algorithms which utilize whole slide images in the field of digital pathology. There are previous studies about histopathology, digital pathology, whole slide imaging and image registration, color staining, data augmentation, and deep learning that are referenced in this study. The goal is to develop a learning-based registration framework specifically for high-resolution histopathology image registration. Different whole slide tissue sample images are used with a resolution of up to 40x magnification. The images are organized into sets of consecutive, differently dyed sections, and the aim is to register the images based on only the visible tissue and ignore the background. Significant structures in the tissue are marked with landmarks. The quality measurements include, for example, the relative target registration error, structural similarity index metric, visual evaluation, landmark-based evaluation, matching points, and image details. These results are comparable and can be used also in the future research and in development of new tools. Moreover, the results are expected to show how the theory and practice are combined in whole slide image registration challenges. DeepHistReg algorithm will be studied to better understand the development of stain color feature augmentation-based image registration tool of this study. Matlab and Aperio ImageScope are the tools to annotate and validate the image, and Python is used to develop the algorithm of this new registration tool. As cancer is globally a serious disease regardless of age or lifestyle, it is important to find ways to develop the systems experts can use while working with patients’ data. There is still a lot to improve in the field of digital pathology and this study is one step toward it.Eri menetelmin värjättyjen virtuaalinäytelasien rekisteröintitekniikka kiintopisteiden validointia hyödyntäen. Tiivistelmä. Yksi tärkeimmistä digitaalipatologian ominaisuuksista on verrata ja fuusioida peräkkäisiä eri menetelmin värjättyjä kudosleikkeitä toisiinsa visuaalisesti. Tällöin keskenään lähes identtiset kuvat kohdistetaan samaan yhteiseen kehykseen, niin sanottuun pohjatotuuteen. Nykyiset näytteiden skannaustyökalut mahdollistavat sellaisten kuvien luonnin, jotka ovat täynnä kerroksittaista tietoa digitalisoiduista näytteistä, tallennettuna erittäin korkean resoluution virtuaalisiin näytelaseihin. Tällä hetkellä on olemassa kuitenkin vain kourallinen automaattisia työkaluja, jotka kykenevät käsittelemään näin valtavia kuvatiedostoja tarkasti hyväksytyin aikarajoin. Tämän työn tarkoituksena on syväoppimista hyväksikäyttäen löytää ratkaisu histopatologisten kuvien rekisteröintiin. Tärkeimpänä osa-alueena on ymmärtää kiintopisteiden validoinnin periaatteet sekä eri väriaineiden augmentoinnin vaikutus. Lisäksi tässä työssä kehitettyä rekisteröintialgoritmia tullaan vertailemaan muihin kirjallisuudessa esitettyihin algoritmeihin, jotka myös hyödyntävät virtuaalinäytelaseja digitaalipatologian saralla. Kirjallisessa osiossa tullaan siteeraamaan aiempia tutkimuksia muun muassa seuraavista aihealueista: histopatologia, digitaalipatologia, virtuaalinäytelasi, kuvantaminen ja rekisteröinti, näytteen värjäys, data-augmentointi sekä syväoppiminen. Tavoitteena on kehittää oppimispohjainen rekisteröintikehys erityisesti korkearesoluutioisille digitalisoiduille histopatologisille kuville. Erilaisissa näytekuvissa tullaan käyttämään jopa 40-kertaista suurennosta. Kuvat kudoksista on järjestetty eri menetelmin värjättyihin peräkkäisiin kuvasarjoihin ja tämän työn päämääränä on rekisteröidä kuvat pohjautuen ainoastaan kudosten näkyviin osuuksiin, jättäen kuvien tausta huomioimatta. Kudosten merkittävimmät rakenteet on merkattu niin sanotuin kiintopistein. Työn laatumittauksina käytetään arvoja, kuten kohteen suhteellinen rekisteröintivirhe (rTRE), rakenteellisen samankaltaisuuindeksin mittari (SSIM), sekä visuaalista arviointia, kiintopisteisiin pohjautuvaa arviointia, yhteensopivuuskohtia, ja kuvatiedoston yksityiskohtia. Nämä arvot ovat verrattavissa myös tulevissa tutkimuksissa ja samaisia arvoja voidaan käyttää uusia työkaluja kehiteltäessä. DeepHistReg metodi toimii pohjana tässä työssä kehitettävälle näytteen värjäyksen parantamiseen pohjautuvalle rekisteröintityökalulle. Matlab ja Aperio ImageScope ovat ohjelmistoja, joita tullaan hyödyntämään tässä työssä kuvien merkitsemiseen ja validointiin. Ohjelmointikielenä käytetään Pythonia. Syöpä on maailmanlaajuisesti vakava sairaus, joka ei katso ikää eikä elämäntyyliä. Siksi on tärkeää löytää uusia keinoja kehittää työkaluja, joita asiantuntijat voivat hyödyntää jokapäiväisessä työssään potilastietojen käsittelyssä. Digitaalipatologian osa-alueella on vielä paljon innovoitavaa ja tämä työ on yksi askel eteenpäin taistelussa syöpäsairauksia vastaan

Data harmonisation for information fusion in digital healthcare: A state-of-the-art systematic review, meta-analysis and future research directions.

Removing the bias and variance of multicentre data has always been a challenge in large scale digital healthcare studies, which requires the ability to integrate clinical features extracted from data acquired by different scanners and protocols to improve stability and robustness. Previous studies have described various computational approaches to fuse single modality multicentre datasets. However, these surveys rarely focused on evaluation metrics and lacked a checklist for computational data harmonisation studies. In this systematic review, we summarise the computational data harmonisation approaches for multi-modality data in the digital healthcare field, including harmonisation strategies and evaluation metrics based on different theories. In addition, a comprehensive checklist that summarises common practices for data harmonisation studies is proposed to guide researchers to report their research findings more effectively. Last but not least, flowcharts presenting possible ways for methodology and metric selection are proposed and the limitations of different methods have been surveyed for future research

Image informatics strategies for deciphering neuronal network connectivity

Brain function relies on an intricate network of highly dynamic neuronal connections that rewires dramatically under the impulse of various external cues and pathological conditions. Among the neuronal structures that show morphologi- cal plasticity are neurites, synapses, dendritic spines and even nuclei. This structural remodelling is directly connected with functional changes such as intercellular com- munication and the associated calcium-bursting behaviour. In vitro cultured neu- ronal networks are valuable models for studying these morpho-functional changes. Owing to the automation and standardisation of both image acquisition and image analysis, it has become possible to extract statistically relevant readout from such networks. Here, we focus on the current state-of-the-art in image informatics that enables quantitative microscopic interrogation of neuronal networks. We describe the major correlates of neuronal connectivity and present workflows for analysing them. Finally, we provide an outlook on the challenges that remain to be addressed, and discuss how imaging algorithms can be extended beyond in vitro imaging studies