Thesis (Ph. D.)--Montana State University--Bozeman, 2007. / Typescript. Chairperson, Graduate Committee: Anne Camper. Includes bibliographical references (leaves 104-113).

The alkali-silica reaction is one of the most damaging chemical reactions taking place in concrete, which can cause fatal damage. ASR originates under following conditions: high moisture (> 80 %), sufficient amount of alkaline ions (Ca2+ , Na+ , K+ ) and use of reactive aggregates (low crystaline or deformed quartz, amorphous SiO2). Reactive aggretates react with high alkaline pore solution and produce hydrofile gels. These gels absorb water and swell. Dilatometric test methods are commonly used to evaluate the reactivity of aggregates. The principle of dilatometric test methods is simple. Mortar or concrete prisms are created in a laboratory, then they are stored in the special environment, which accelerates the inception of ASR. The creation and expansion of alkali-silica gels cause prism's length changes. The major goal of this diploma thesis was to evaluate the alkali-silica reactivity potential of quartz-rich rocks using microscopic (polarizing microscopy, scanning electron microscopy combined with SEM/BSE image analysis) and dilatometric (ASTM C1260, RILEM AAR-4.1) methods. Rocks were assessed as reactive, potentially reactive and non-reactive by the ASTM C1260 method. The reactivity of aggregates was connected with the amount of cryptocrystaline matrix, grain size, shape of grain boundaries,...

Fluorescence microscopy is an indispensable tool for biology to study the spatio-temporal dynamics of cells, tissues, and developing organisms. Modern imaging modalities, such as light-sheet microscopy, are able to acquire large three- dimensional volumes with high spatio-temporal resolution for many hours or days, thereby routinely generating Terabytes of image data in a single experiment. The quality of these images, however, is limited by the optics of the microscope, the signal-to-noise ratio of acquisitions, the photo-toxic effects of illumination, and the distortion of light by the sample. Additionally, the serial operation mode of most microscopy experiments, where large data sets are first acquired and only afterwards inspected and analyzed, excludes the possibility to optimize image quality during acquisition by automatically adapting the microscope parameters. These limits make certain observations difficult or impossible, forcing trade-offs between imaging speed, spatial resolution, light exposure, and imaging depth. This thesis is concerned with addressing several of these challenges with computational methods. First, I present methods for visualizing and processing the volumetric data from a microscope in real-time, i.e. at the acquisition rate of typical experiments, which is a prerequisite for the development of adaptive microscopes. I propose a low-discrepancy sampling strategy that enables the seamless display of large data sets during acquisition, investigate real-time compatible denoising, convolution, and deconvolution methods, and introduce a low-rank decomposition strategy for common deblurring tasks. Secondly, I propose a computational tractable method to simulate the interaction of light with realistically large biological tissues by combining a GPU-accelerated beam propagation method with a novel multiplexing scheme. I demonstrate that this approach enables to rigorously simulate the wave-optical image formation in light-sheet microscopes, to numerically investigate correlative effects in scattering tissues, and to elucidate the optical properties of the inverted mouse retina. Finally, I propose a data-driven restoration approach for fluorescence microscopy images based on convolutional neural networks (Care) that leverages sample and imaging specific prior knowledge. By demonstrating the superiority of this approach when compared to classical methods on a variety of problems, ranging from restoration of high quality images from low signal-to-noise-ratio acquisitions, to projection of noisy developing surface, isotropic recovery from anisotropic volumes, and to the recovery of diffraction-limited structures from widefield images alone, I show that Care is a flexible and general method to solve fundamental restoration problems in fluorescence microscopy.

Fluorescence microscopy is an invaluable technique for studying the intricate process of organism development. The acquisition process, however, is associated with the fundamental trade-off between the quality and reliability of the acquired data. On one hand, the goal of capturing the development in its entirety, often times across multiple spatial and temporal scales, requires extended acquisition periods. On the other hand, high doses of light required for such experiments are harmful for living samples and can introduce non-physiological artifacts in the normal course of development. Conventionally, a single set of acquisition parameters is chosen in the beginning of the acquisition and constitutes the experimenter’s best guess of the overall optimal configuration within the aforementioned trade-off. In the paradigm of adaptive microscopy, in turn, one aims at achieving more efficient photon budget distribution by dynamically adjusting the acquisition parameters to the changing properties of the sample. In this thesis, I explore the principles of adaptive microscopy and propose a range of improvements for two real imaging scenarios. Chapter 2 summarizes the design and implementation of an adaptive pipeline for efficient observation of the asymmetrically dividing neurogenic progenitors in Zebrafish retina. In the described approach the fast and expensive acquisition mode is automatically activated only when the mitotic cells are present in the field of view. The method illustrates the benefits of the adaptive acquisition in the common scenario of the individual events of interest being sparsely distributed throughout the duration of the acquisition. Chapter 3 focuses on computational aspects of segmentation-based adaptive schemes for efficient acquisition of the developing Drosophila pupal wing. Fast sample segmentation is shown to provide a valuable output for the accurate evaluation of the sample morphology and dynamics in real time. This knowledge proves instrumental for adjusting the acquisition parameters to the current properties of the sample and reducing the required photon budget with minimal effects to the quality of the acquired data. Chapter 4 addresses the generation of synthetic training data for learning-based methods in bioimage analysis, making them more practical and accessible for smart microscopy pipelines. State-of-the-art deep learning models trained exclusively on the generated synthetic data are shown to yield powerful predictions when applied to the real microscopy images. In the end, in-depth evaluation of the segmentation quality of both real and synthetic data-based models illustrates the important practical aspects of the approach and outlines the directions for further research.

The goal of the thesis is to find the way how to apply use-wear analysis as well as functional analysis to archaeological assemblages of selected artefacts from Central European archaeological contexts, namely lithics (chipped stone artefacts) and obtain the greatest potential from the analysis. Thesis consists of: current state in the field of functional studies research worldwide, method of use-wear application in connection with material science knowledge, especially tribology. Use-wear analysis is applied to the selected lithic artefacts from Paleolithic to Eneolithic Periods. The results of this work are based on the following microscopic approaches: low power approach (LPA), high power approach (HPA), scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). There are described and stated differences in potential of above mentioned approaches in connection with specific archaeological artefacts (assemblages of artefacts). The question answered in the conclusion is how to apply the above mentioned methodological approaches in application to various archaeological materials (period, number, context etc.) to obtain the greatest informational potential from the material analysed. Work is concluded with specific terminology from the field of tribology and use-wear analysis...

<div>Recent advances in optical microscopy have enabled biologists collect fluorescence microscopy volumes cellular and subcellular structures of living tissue. This results in collecting large datasets of microscopy volume and needs image processing aided automated quantification method. To quantify biological structures a first and fundamental step is segmentation. Yet, the quantitative analysis of the microscopy volume is hampered by light diffraction, distortion created by lens aberrations in different directions, complex variation of biological structures. This thesis describes several proposed segmentation methods to identify various biological structures such as nuclei or tubules observed in fluorescence microscopy volumes. To achieve nuclei segmentation, multiscale edge detection method and 3D active contours with inhomogeneity correction method are used for segmenting nuclei. Our proposed 3D active contours with inhomogeneity correction method utilizes 3D microscopy volume information while addressing intensity inhomogeneity across vertical and horizontal directions. To achieve tubules segmentation, ellipse model fitting to tubule boundary method and convolutional neural networks with inhomogeneity correction method are performed. More specifically, ellipse fitting method utilizes a combination of adaptive and global thresholding, potentials, z direction refinement, branch pruning, end point matching, and boundary fitting steps to delineate tubular objects. Also, the deep learning based method combines intensity inhomogeneity correction, data augmentation, followed by convolutional neural networks architecture. Moreover, this thesis demonstrates a new deconvolution method to improve microscopy image quality without knowing the 3D point spread function using a spatially constrained cycle-consistent adversarial networks. The results of proposed methods are visually and numerically compared with other methods. Experimental results demonstrate that our proposed methods achieve better performance than other methods for nuclei/tubules segmentation as well as deconvolution.</div>

Mathematical models of signal transduction pathways are characterized by a large number of proteins and uncertain parameters, yet only a limited amount of quantitative data is available. The dissertation addresses this problem using two different approaches: the first approach deals with a model simplification procedure for signaling pathways that reduces the model size but retains the physical interpretation of the remaining states, while the second approach deals with creating rich data sets by computing transcription factor profiles from fluorescent images of green-fluorescent-protein (GFP) reporter cells. For the first approach a model simplification procedure for signaling pathway models is presented. The technique makes use of sensitivity and observability analysis to select the retained proteins for the simplified model. The presented technique is applied to an IL-6 signaling pathway model. It is found that the model size can be significantly reduced and the simplified model is able to adequately predict the dynamics of key proteins of the signaling pathway. An approach for quantitatively determining transcription factor profiles from GFP reporter data is developed as the second major contribution of this work. The procedure analyzes fluorescent images to determine fluorescence intensity profiles using principal component analysis and K-means clustering, and then computes the transcription factor concentration from the fluorescence intensity profiles by solving an inverse problem involving a model describing transcription, translation, and activation of green fluorescent proteins. Activation profiles of the transcription factors NF-κB, nuclear STAT3, and C/EBPβ are obtained using the presented approach. The data for NF-κB is used to develop a model for TNF-α signal transduction while the data for nuclear STAT3 and C/EBPβ is used to verify the simplified IL-6 model. Finally, an approach is developed to compute the distribution of transcription factor profiles among a population of cells. This approach consists of an algorithm for identifying individual fluorescent cells from fluorescent images, and an algorithm to compute the distribution of transcription factor profiles from the fluorescence intensity distribution by solving an inverse problem. The technique is applied to experimental data to derive the distribution of NF-κB concentrations from fluorescent images of a NF-κB GFP reporter system.

Abstract Biology and medicine rely heavily on images to understand how the body functions, for diagnosing diseases and to test the effects of treatments. In recent decades, microscopy has experienced rapid improvements, enabling imaging of fixed and living cells at higher resolutions and frame rates, and deeper inside the biological samples. This has led to rapid growth in the image data. Automated methods are needed to quantitatively analyze these huge datasets and find statistically valid patterns. Cell segmentation and tracking is critical for automated analysis, yet it is a challenging problem due to large variations in cell shapes and appearances caused by various factors, including cell type, sample preparation and imaging setup. This thesis proposes novel methods for segmentation and tracking of cells, which rely on machine learning based approaches to improve the performance, generalization and reusability of automated methods. Cell proposals are used to efficiently exploit spatial and temporal context for resolving detection ambiguities in high-cell-density regions, caused by weak boundaries and deformable shapes of cells. This thesis presents two cell proposal methods: the first method uses multiple blob-like filter banks for detecting candidates for round cells, while the second method, Cell Proposal Network (CPN), uses convolutional neural networks to learn the cell shapes and appearances, and can propose candidates for cells in a wide variety of microscopy images. CPN first regresses cell candidate bounding boxes and their scores, then, it segments the regions inside the top ranked boxes to obtain cell candidate masks. CPN can be used as a general cell detector, as is demonstrated by training a single model to segment images from histology, fluorescence and phase-contrast microscopy. This work poses segmentation and tracking as proposal selection problems, which are solved optimally using integer linear programming or approximately using iterative shortest cost path search and non-maximum suppression. Additionally, this thesis presents a method which utilizes graph-cuts and an off-the-shelf edge detector to accurately segment highly deformable cells. The main contribution of this thesis is a cell tracking method which uses CPN to propose cell candidates, represents alternative tracking hypotheses using a graphical model, and selects the globally optimal sub-graph providing cell tracks. It achieves state-of-the-art tracking performance on multiple public benchmark datasets from both phase-contrast and fluorescence microscopy containing cells of various shapes and appearances. / Tiivistelmä Biologia ja lääketiede nojaavat vahvasti kuvatietoon solujen ja kehon toimintojen ymmärtämiseksi sairauksien diagnostiikassa ja hoitojen vaikutusten seuraamisessa. Viime vuosikymmeninä mikroskopiassa on tapahtunut nopeaa teknistä kehitystä, mikä on mahdollistanut elävien solujen kuvantamisen tarkemmin, nopeammin sekä syvemmältä automatisoidusti useasta näytteestä. Tämä taas on johtanut kuvadatan nopeaan kasvuun ja suurempaan määrään biologisia kysymyksiä, joihin voidaan vastata. Kuvadatan räjähdysmäisen kasvun vuoksi kaikkia tuloksia ei voida enää tulkita pelkästään ihmistyövoimaa käyttämällä, mikä on johtanut tarpeeseen kehittää automaattisia menetelmiä analysoimaan kvantitatiivisesti suuria datajoukkoja ja löytämään tilastollisesti kelvollisia malleja. Solujen erottaminen niiden ympäristöstä ja toisista soluista (segmentointi) ja solujen seuranta ovat kriittisiä alkuvaiheen osia onnistuneessa automaattisessa analyysissä. Automaattisten menetelmien kehittämisessä solusegmentointi on kuitenkin osoittautunut hyvin haastavaksi ongelmaksi solujen muodon ja ulkonäön suurten muutosten vuoksi solutyypistä, näytteen valmistelusta ja kuvantamisjärjestelmästä johtuen. Tämä väitöskirja esittää uusia menetelmiä solujen segmentointiin ja seurantaan keskittyen koneoppimiseen perustuviin lähestymistapoihin, jotka parantavat automaattisten menetelmien suorituskykyä ja uudelleenkäytettävyyttä. Spatiaalista ja ajallista kontekstia tehokkaasti hyödyntäviä soluehdotelmia käytetään ratkaisemaan solujen heikosti erottuvista reunoista ja joustavista muodoista johtuvaa solujen muodon monitulkintaisuutta erityisesti silloin kun tutkittava solutiheys on suuri. Tämä väitöskirja esittää kaksi menetelmää soluehdotelmille; ensimmäinen menetelmä käyttää useita läikkätyyppisiä suodatinpankkeja ilmaisemaan kandidaatteja pyöreänmuotoisille soluille, kun taas toinen menetelmä nimeltään soluehdotelmaverkko (Cell Proposal Network, CPN) käyttää konvoluutionaalisia neuroverkkoja oppiakseen tunnistamaan solut niiden muodon sekä ulkonäön perusteella erityyppisissä mikroskooppikuvissa. CPN regressoi ensin solukandidaatteja ympäröivät suorakaiteet ja niiden pistemäärän, jonka jälkeen se segmentoi alueet parhaiten sijoittuneiden suorakaiteiden joukosta tuottaen solukandidaattimaskit. CPN:ää voidaan mahdollisesti käyttää yleisenä soluilmaisimena erityyppisilla kuvantamistekniikoilla tuotetuissa kuvissa mukaan lukien histologisen valo-, fluoresenssi- ja vaihekontrastimikroskooppian. Väitöskirja esittää solujen segmentoinnin ja seurannan soluehdotelmien valintaongelmina, mitkä ratkaistaan joko optimaalisesti käyttämällä kokonaislukuoptimointia tai likimääräisesti käyttämällä iteratiivista lyhimmän kustannuspolun hakua sekä ei-maksimien vaimennusta. Tämä väitöskirja esittää myös verkon leikkaukseen (graph cut) perustuvan menetelmän, mikä hyödyntää valmiiksi saatavilla olevaa reunanilmaisinta segmentoimaan tarkasti muotoaan voimakkaasti muuttavia soluja. Väitöskirjatutkimuksen keskeinen tulos on uusi solujen seurantamenetelmä, mikä käyttää CPN:ää solukandidaattien ehdottamiseen, esittää vaihtoehtoiset seurantahypoteesit verkkomallia hyödyntämällä, ja valitsee globaalisti optimaalisen aliverkon solujen kulkemille reitille. Verrattuna useisiin muihin julkisesti saatavilla oleviin kuva-analyysiohjelmistoihin tässä väitöskirjassa kehitetyt menetelmät olivat suorituskyvyltään parhaita vaihekontrasti- ja fluoresenssimikroskopialla tuotettujen kuva-aineistojen analyyseissa, joissa solujen ulkomuoto oli hyvin vaihteleva.

It is known that fission yeast Schizosaccharomyces pombe maintains its nuclear envelope during mitosis and it undergoes an interesting shape change during cell division - from a spherical via an ellipsoidal and a peanut-like to a dumb-bell shape. However, the biomechanical system behind this amazing transformation is still not understood. What we know is, that the shape must change due to forces acting on the membrane surrounding the nucleus and the microtubule based mitotic spindle is thought to play a key role. To estimate the locations and directions of the forces, the shape of the nucleus was recorded by confocal light microscopy. But such data is often inhomogeneously labeled with gaps in the boundary, making classical segmentation impractical. In order to accurately determine the shape we developed a global parametric shape description method, based on a Fourier coordinate expansion. The method implicitly assumes a closed and smooth surface. We will calculate the geometrical properties of the 2-dimensional shape and extend it to 3-dimensional properties, assuming rotational symmetry. Using a mechanical model for the lipid bilayer and the so called Helfrich-Canham free energy we want to calculate the minimum energy shape while respecting system-specific constraints to the surface and the enclosed volume. Comparing it with the observed shape leads to the forces. This provides the needed research tools to study forces based on images.