How automated image analysis techniques help scientists in species identification and classification?

Identification of taxonomy at a specific level is time consuming and reliant upon expert ecologists. Hence the demand for automated species identification incre­ased over the last two decades. Automation of data classification is primarily focussed on images while incorporating and analysing image data has recently become easier due to developments in computational technology. Research ef­forts on identification of species include specimens’ image processing, extraction of identical features, followed by classifying them into correct categories. In this paper, we discuss recent automated species identification systems, mainly for categorising and evaluating their methods. We reviewed and compared different methods in step by step scheme of automated identification and classification systems of species images. The selection of methods is influenced by many variables such as level of classification, number of training data and complexity of images. The aim of writing this paper is to provide researchers and scientists an extensive background study on work related to automated species identification, focusing on pattern recognition techniques in building such systems for biodiversity studies. (Folia Morphol 2018; 77, 2: 179–193

Automated Computational Techniques for High-throughput Image Analysis of Skin Structure

Biological image processing and analysis are concerned with enhancing and quantifying features that reflect different pathological states, based on the use of combinations of image processing algorithms. The integration of image processing and analysis techniques to evaluate and assess skin integrity in both human and mouse models is a major theme in this thesis. More specifically, this thesis describes computational systems for high-throughput analysis of skin tissue section images and non-invasive imaging techniques. As the skin is a largest organ in the mammalian body, and is complex in structure, manual quantification and analysis a hard task for the observer to determine an objective result, and furthermore, the analysis is complex in terms of accuracy and time taken. To look at the gross morphology of the skin, I developed high throughput analysis based on an adaptive active contour model to isolate the skin layers and provide quantification methods. This was utilised in a study to evaluate cutaneous morphology in 475 knockout mouse lines provided by the Mouse Genetics Project (MGP) pipeline, that was generated by the Wellcome Trust Sanger Institute (WTSI). This is a major international initiative to provide both functional annotation of the mammalian genome and insight into the genetic basis of disease. I found 53 interesting adipocyte phenotypes, 18 interesting dermal phenotypes and 3 interesting epidermal phenotypes. I also focussed on the analysis of collagen in the dermis of skin images in several ways. For collagen structure analysis, I developed a combined system of Gabor filtering and Fast Fourier Transform FFT. This analysis allowed the detection of subtle changes in collagen organisation. Using similar images, I also measured collagen bundle thickness by computing the maximum frequency of the FFT power spectrum. To assess collagen dynamics, I developed k-means clustering for segmentation based on colour distribution. The use of this approach allowed the measurement of dermal degradation with age and disease, which was not possible by existing means. Obtaining human skin material to facilitate the drug discovery and development process is not an easy task. The manipulation, monitoring and cost of human subjects makes the use of mouse models more suitable for high-throughput screening. Therefore, I have evaluated skin integrity from mouse tissue rather than human skin, however, mouse skin is thinner than human skin and many morphological features are easier to visualise in human skin, which has implications for analysis. Skin moulds can be used to create an impression of the skin surface. Changes in texture of skin can reflect skin conditions. I developed a skin surface structure analysis system to measure the degree of change in texture of the human skin surface. The alterations detected in texture parameters in skin mould impressions reflected changes caused by sun exposure, ageing and many other clinical parameters. I compared my analysis with the existing Beagley-Gibson scoring system to find correlations between automated and manual analysis to inform a decision on the use of optimal methods. By removing subjectivity of manual methods, I was able to develop a robust system to evaluate, for example, damage resulting from UV exposure. My experimental analysis indicated that techniques developed in this thesis were able to analyse both histological samples and skin surface images in high-throughput experiments. They could, therefore, make a contribution to biological image analysis by providing accurate results to help clinical decision making, and facilitate biological laboratory experiments to improve the quality of research in this field, and save time. Overall, my thesis demonstrated that accurate analysis of the skin to gain meaningful biological information requires an automated system that can achieve feature extraction, quantification, analysis and decision making to find interesting phenotypes and abnormalities. This will help the evaluation of the effects of a specific treatment, and answer many biological questions in fields of cosmetic dermatology and drug discovery, and improve our understanding of the genetic basis of disease

New algorithms for the analysis of live-cell images acquired in phase contrast microscopy

La détection et la caractérisation automatisée des cellules constituent un enjeu important dans de nombreux domaines de recherche tels que la cicatrisation, le développement de l'embryon et des cellules souches, l’immunologie, l’oncologie, l'ingénierie tissulaire et la découverte de nouveaux médicaments. Étudier le comportement cellulaire in vitro par imagerie des cellules vivantes et par le criblage à haut débit implique des milliers d'images et de vastes quantités de données. Des outils d'analyse automatisés reposant sur la vision numérique et les méthodes non-intrusives telles que la microscopie à contraste de phase (PCM) sont nécessaires. Comme les images PCM sont difficiles à analyser en raison du halo lumineux entourant les cellules et de la difficulté à distinguer les cellules individuelles, le but de ce projet était de développer des algorithmes de traitement d'image PCM dans Matlab® afin d’en tirer de l’information reliée à la morphologie cellulaire de manière automatisée. Pour développer ces algorithmes, des séries d’images de myoblastes acquises en PCM ont été générées, en faisant croître les cellules dans un milieu avec sérum bovin (SSM) ou dans un milieu sans sérum (SFM) sur plusieurs passages. La surface recouverte par les cellules a été estimée en utilisant un filtre de plage de valeurs, un seuil et une taille minimale de coupe afin d'examiner la cinétique de croissance cellulaire. Les résultats ont montré que les cellules avaient des taux de croissance similaires pour les deux milieux de culture, mais que celui-ci diminue de façon linéaire avec le nombre de passages. La méthode de transformée par ondelette continue combinée à l’analyse d'image multivariée (UWT-MIA) a été élaborée afin d’estimer la distribution de caractéristiques morphologiques des cellules (axe majeur, axe mineur, orientation et rondeur). Une analyse multivariée réalisée sur l’ensemble de la base de données (environ 1 million d’images PCM) a montré d'une manière quantitative que les myoblastes cultivés dans le milieu SFM étaient plus allongés et plus petits que ceux cultivés dans le milieu SSM. Les algorithmes développés grâce à ce projet pourraient être utilisés sur d'autres phénotypes cellulaires pour des applications de criblage à haut débit et de contrôle de cultures cellulaires.Automated cell detection and characterization is important in many research fields such as wound healing, embryo development, immune system studies, cancer research, parasite spreading, tissue engineering, stem cell research and drug research and testing. Studying in vitro cellular behavior via live-cell imaging and high-throughput screening involves thousands of images and vast amounts of data, and automated analysis tools relying on machine vision methods and non-intrusive methods such as phase contrast microscopy (PCM) are a necessity. However, there are still some challenges to overcome, since PCM images are difficult to analyze because of the bright halo surrounding the cells and blurry cell-cell boundaries when they are touching. The goal of this project was to develop image processing algorithms to analyze PCM images in an automated fashion, capable of processing large datasets of images to extract information related to cellular viability and morphology. To develop these algorithms, a large dataset of myoblasts images acquired in live-cell imaging (in PCM) was created, growing the cells in either a serum-supplemented (SSM) or a serum-free (SFM) medium over several passages. As a result, algorithms capable of computing the cell-covered surface and cellular morphological features were programmed in Matlab®. The cell-covered surface was estimated using a range filter, a threshold and a minimum cut size in order to look at the cellular growth kinetics. Results showed that the cells were growing at similar paces for both media, but their growth rate was decreasing linearly with passage number. The undecimated wavelet transform multivariate image analysis (UWT-MIA) method was developed, and was used to estimate cellular morphological features distributions (major axis, minor axis, orientation and roundness distributions) on a very large PCM image dataset using the Gabor continuous wavelet transform. Multivariate data analysis performed on the whole database (around 1 million PCM images) showed in a quantitative manner that myoblasts grown in SFM were more elongated and smaller than cells grown in SSM. The algorithms developed through this project could be used in the future on other cellular phenotypes for high-throughput screening and cell culture control applications

3D Object Recognition Based On Constrained 2D Views

The aim of the present work was to build a novel 3D object recognition system capable of classifying man-made and natural objects based on single 2D views. The approach to this problem has been one motivated by recent theories on biological vision and multiresolution analysis. The project's objectives were the implementation of a system that is able to deal with simple 3D scenes and constitutes an engineering solution to the problem of 3D object recognition, allowing the proposed recognition system to operate in a practically acceptable time frame. The developed system takes further the work on automatic classification of marine phytoplank- (ons, carried out at the Centre for Intelligent Systems, University of Plymouth. The thesis discusses the main theoretical issues that prompted the fundamental system design options. The principles and the implementation of the coarse data channels used in the system are described. A new multiresolution representation of 2D views is presented, which provides the classifier module of the system with coarse-coded descriptions of the scale-space distribution of potentially interesting features. A multiresolution analysis-based mechanism is proposed, which directs the system's attention towards potentially salient features. Unsupervised similarity-based feature grouping is introduced, which is used in coarse data channels to yield feature signatures that are not spatially coherent and provide the classifier module with salient descriptions of object views. A simple texture descriptor is described, which is based on properties of a special wavelet transform. The system has been tested on computer-generated and natural image data sets, in conditions where the inter-object similarity was monitored and quantitatively assessed by human subjects, or the analysed objects were very similar and their discrimination constituted a difficult task even for human experts. The validity of the above described approaches has been proven. The studies conducted with various statistical and artificial neural network-based classifiers have shown that the system is able to perform well in all of the above mentioned situations. These investigations also made possible to take further and generalise a number of important conclusions drawn during previous work carried out in the field of 2D shape (plankton) recognition, regarding the behaviour of multiple coarse data channels-based pattern recognition systems and various classifier architectures. The system possesses the ability of dealing with difficult field-collected images of objects and the techniques employed by its component modules make possible its extension to the domain of complex multiple-object 3D scene recognition. The system is expected to find immediate applicability in the field of marine biota classification