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Biophysical interpretation of quantitative phase image
Štrbková, Lenka ; Kozubek,, Michal (referee) ; Hoppe, Andreas (referee) ; Chmelík, Radim (advisor)
Práce se zabývá interpretací kvantitativního fázového zobrazení pomocí techniky koherencí řízené holografické mikroskopie. Vzhledem k tomu, že tato technika generuje velké množství kvantitativních fázových obrazů o nezanedbatelné velikosti, manuální analýza by byla časově náročná a neefektivní Za účelem urychlení analýzy obrazů získaných pomocí koherencí řízené holografické mikroskopie je v této práci navržena metodika automatizované interpretace kvantitativních fázových obrazů pomocí strojového učení s učitelem. Kvantitativní fázové obrazy umožňují extrakci parametrů charakterizujících distribuci suché hmoty v buňce a poskytují tak cennou informaci o buněčném chování. Cílem této práce je navrhnout metodologii pro automatizovanou klasifikaci buněk při využití této kvantitativní informace jak ze statických, tak z časosběrných kvantitativních fázových obrazů. Navržená metodika byla testována v experimentech s živými buňkami, jimiž byla vyhodnocena výkonnost klasifikace a významnost parametrů získaných z kvantitativních fázových obrazů.
Methods of Detection, Segmentation and Classification of Difficult to Define Bone Tumor Lesions in 3D CT Data
Chmelík, Jiří ; Flusser,, Jan (referee) ; Kozubek, Michal (referee) ; Jan, Jiří (advisor)
The aim of this work was the development of algorithms for detection segmentation and classification of difficult to define bone metastatic cancerous lesions from spinal CT image data. For this purpose, the patient database was created and annotated by medical experts. Successively, three methods were proposed and developed; the first of them is based on the reworking and combination of methods developed during the preceding project phase, the second method is a fast variant based on the fuzzy k-means cluster analysis, the third method uses modern machine learning algorithms, specifically deep learning of convolutional neural networks. Further, an approach that elaborates the results by a subsequent random forest based meta-analysis of detected lesion candidates was proposed. The achieved results were objectively evaluated and compared with results achieved by algorithms published by other authors. The evaluation was done by two objective methodologies, technical voxel-based and clinical object-based ones. The achieved results were subsequently evaluated and discussed.
The analysis of limits for multimode fibre imaging
Štolzová, Hana ; Kozubek, Michal (referee) ; Dostál, Zbyněk (advisor)
Multimódová vlákna jsou zobrazovacím prostředkem s významným potenciálem v in-vivo mikroendoskopii. V poslední době tato metoda zaznamenala velký rozvoj, a to díky zdokonalování výpočetní a jiné techniky, například prostorové modulace světla. Cílem této práce bylo nalézt specifické limity zobrazování multimódovými vlákny a představit jejich počítačovou simulaci. Byl zkoumán vliv způsobu osvětlení optického systému obsahujícího multimódové vlákno na jeho schopnost fokusace a zobrazování. Analýzou dat získaných ze simulací a experimentu bylo zjištěno, že různá míra omezení Gaussovského svazku a plnění apertury multimódového vlákna má za následek významnou změnu zobrazovacích schopností systému. Při pozorování kvality fokusace bylo zjištěno, že nejlépe se projevují svazky málo omezené aperturou vlákna. Tento fakt byl potvrzen i experimentálním měřením. Zobrazování za použití svazků s podobnými hodnotami omezení (50%) projevovalo i nejlepší schopnost přenosu kontrastu. Avšak při analýze rozlišení dvou bodových objektů se jako nejvhodnější projevily svazky významně přeplňující numerickou aperturu vlákna, 100% a více. Přítomnost tohoto rozdílu poukazuje na skutečnost, že multimódové vlákno není zcela náhodné médium, ale propagace světla skrz multimódové vlákno projevuje znaky závislosti na vnějších zobrazovacích podmínkách, jako je například změna omezení osvětlovacího svazku. V této práci bylo představeno několik způsobů vyhodnocení kvality zobrazování pomocí multimódového vlákna. Každé z těchto kritérií podalo dílčí charakteristiku chování optického systému obsahujícího multimódové optické vlákno. Jednotlivé výsledky se neshodují na jednom konkrétním řešení a nutí osobu využívající zobrazovací systém obsahující multimódové vlákno ke zvážení několika aspektů, a to v jakém prostředí bude daný optický systém využívat a který parametr kvality zobrazení bude považovat za nejdůležitější.
Confocal module for the Coherence Controlled Holographic Microscope
Kubátová, Eva ; Kozubek, Michal (referee) ; Dostál, Zbyněk (advisor)
The Coherence Controlled Holographic Microscope (CCHM) was developed at BUT Brno for a quantitative phase imaging of living cells. Nowadays it ocurres that its imaging properties are enhanced by the use of additional modules. In the present the microscope is equipped with the epifluorescence module, which allows observation of fluorescently marked living cells. This thesis is going to follow up on the development of this module and is going to extend its options by confocal imaging. The disadvantage of current multi-channel confocal microscopes is a mechanical rotation of the Nipkow discs, which causes undesired mechanical vibrations. That is why in this thesis it is replaced by Digital Micromirror Device. With its use was developed optical system of the whole confocal model, whose correct funcion was simulated in optical CAD. The experimentally verified prototype serves to test the imaging properties. On this basis is designed an application idea of the fluorescence confocal module, which will be possible to connect to the CCHM microscope.
Coherence-controlled holographic microscope in cell's life cycle research
Křížová, Aneta ; Kozubek,, Michal (referee) ; Uhlířová, Hana (advisor)
The goal of this diploma thesis was using of a coherence-controlled holographic microscope in cell’s life research. A brief history of interference microscopy and it’s applications in biology is described. Also other microscopy techniques routinely used for transparent objects imaging are mentioned and the biology of cell’s life cycle briefly explained. Characteristics describing the shape of a cell were proposed and tested with respect to identification of particular phases of its life cycle. The method of dynamic phase differences was modified in order to distinguish the internal motion of cell’s mass from the movement of the whole cell. Selected characteristics were used to evaluate observations carried out with the holographic microscope and the possibilities of their further applications were depicted. In conclusion, obtained findings were summarized and modifications of microscope construction as well as data-processing software were suggested.
Microscopy of Time Variable Biologic Objects
Uhlířová, Hana ; Kozubek, Michal (referee) ; Peychl,, Jan (referee) ; Chmelík, Radim (advisor)
The subject of the PhD thesis is the application of a transmission digital holographic microscope (DHM) which was designed and constructed in the Laboratory of optical microscopy at the IPE BUT for the research of live cells dynamics. First part of the work is concerned with theoretical description of the microscope imaging properties dependent on the coherence of illumination. It is supplemented with experiments of imaging of a model and a real biological specimen. The following part describes construction modifications and innovations of the microscope and its equipment that enabled the utilization of the microscope for live cells observations. In the experimental part the methodology of live cells preparation and DHM imaging was worked out. The methodology was verified by the observation of cell dynamics during an apoptosis induced by the cytostaticum cis-platinum. Further experiments examined the dynamics of live cells in standard conditions and during a deprivation stimulus. A novel method of holographically reconstructed phase, named \uva{dynamic phase differences}, was set up to evaluate quantitative changes of cell mass distribution during the experiments. Depending on the degree of malignancy and density of cell outgrowth, various schemes of cancer cells behaviour during a specific reaction were revealed using this method. For the quantitative analysis of the DHM phase imaging, a suitable statistical characteristic and an interpretation of the measured data were proposed. Both of them were successfully applied for the comparison of cell motility of two cell types: parental and progeny cell lines. On the basis of the proposed processing, hypotheses describing the reaction mechanism of tumour cells to stress life conditions were established. In the conclusions we summarize our findings and suggestions for the construction and the applications of a new generation of the transmission DHM.
Deep Learning for Virtual Patient-Specific Skull Modelling and Reconstruction
Kodym, Oldřich ; Kozubek, Michal (referee) ; Egger, Bernhard (referee) ; Herout, Adam (advisor)
Segmentace lebky ze 3D pacientských dat a virtuální rekonstrukce tvaru lebek s defekty jsou nejnáročnějšími kroky potřebnými pro tvorbu lebečních modelů na míru pacienta. Tyto modely jsou v kranioplastice využívány pro plánování operací, poučení pacienta a design implantátů na míru, avšak jejich využitelnost je v současnosti limitována množstvím manuální práce potřebné pro dosažení dostatečné kvality virtuálních modelů.  Tato teze má za cíl zefektivnění tohoto virtuálního pracovního postupu s využitím metod hlubokého učení. Teze popisuje klinickou motivaci a současnou výzkumnou literaturu v oblasti automatizace virtuální kranioplastiky. Dále navrhuje nové řešení sestávajicí z metody automatické segmentace lebky založené na kombinaci konvoluční neuronové sítě a algoritmu graph-cut a metody automatické rekonstrukce lebky založené na kaskádě konvolučních sítí. Obě tyto komponenty demonstrují přesnost na úrovni vědeckého stavu poznání. Dále tato práce cílí na zvýšení reprodukovatelnosti výzkumu lebečních rekonstrukcí poskytnutím strukturovaného syntetického datasetu pro vývoj a srovnávání automatických metod. Hlavním cílem této práce je využitelnost v klinické praxi. Zatímco navržená metoda segmentace lebek je již v klinické praxi využívána, integrace automatické virtuální rekonstrukce lebky představuje několik dalších překážek, jako nízká tolerance k nepřesnostem ve tvaru okolo hranice defektu. Tato práce proto také navrhuje rozšíření metody rekonstrukce lebky, které umožňuje její adaptaci na cílovou populaci a typ kraniálních implantátů, který se může mezi jednotlivými klinickými pracovištěmi lišit. Výsledky vyhodnocení experta ukazují, že výstupy této metody dosahují dostatečné kvality pro implementaci do klinické praxe společně s metodou segmentace.
Acquisition, Modeling and Signal Processing in Ultrasound Perfusion Imaging
Mézl, Martin ; Kozubek, Michal (referee) ; Flusser,, Jan (referee) ; Jiřík, Radovan (advisor)
This work deals with possibilities of ultrasound perfusion analysis for the absolute quantification of perfusion parameters. In the theoretical part of this work are discussed possibilities of using of the ultrasound contrast agents and approaches for the perfusion analysis. New methods for the perfusion analysis are suggested and tested in the practical part of this work. The methods are based on convolutional model in which the concentration of the contrast agent is modeled as aconvolution of the arterial input function and the tissue residual function. The feasibility of these methods for the absolute quantification of perfusion parameters is shown on data from phantom studies, simulations and also preclinical and clinical studies. The software for the whole process of the perfusion analysis was developed for using in hospitals.
COHERENCE-CONTROLLED HOLOGRAPHIC MICROSCOPE
Kolman, Pavel ; Křupka, Ivan (referee) ; Kozubek, Michal (referee) ; Chmelík, Radim (advisor)
ransmitted-light coherence-controlled holographic microscope (CCHM) based on an off-axis achromatic and space-invariant interferometer with a diffractive beamsplitter has been designed, constructed and tested. It is capable to image objects illuminated by light sources of arbitrary degree of temporal and spatial coherence. Off-axis image-plane hologram is recorded and the image complex amplitude (intensity and phase) is reconstructed numerically using fast Fourier transform algorithms. Phase image represents the optical path difference between the object and the reference arms caused by presence of an object. Therefore, it is a quantitative phase contrast image. Intensity image is confocal-like. Optical sectioning effect induced by an extended, spatial incoherent light source is equivalent to a conventional confocal image. CCHM is therefore capable to image objects under a diffusive layer or immersed in a turbid media. Spatial and temporal incoherence of illumination makes the optical sectioning effect stronger compared to a confocal imaging process. Object wave reconstruction from the only one recorded interference pattern ensures high resistance to vibrations and medium or ambience fluctuations. The frame rate is not limited by any component of the optical setup. Only the detector and computer speeds limit the frame rate. CCHM therefore allows observation of rapidly varying phenomena. CCHM makes the ex-post numerical refocusing possible within the coherence volume. Coherence degree of the light source in CCHM can be adapted to the object and to the required image properties. More coherent illumination provides wider range of numerical refocusing. On the other hand, a lower degree of coherence makes the optical sectioning stronger, i.e. the optical sections are thiner, it reduces coherence-noise and it makes it possible to separate the ballistic light. In addition to the ballistic light separation, CCHM enables us to separate the diffused light. Multi-colour-light
Methods of Segmentation and Identification of Deformed Vertebrae in 3D CT Data of Oncological Patients
Jakubíček, Roman ; Flusser, Jan (referee) ; Kozubek, Michal (referee) ; Jan, Jiří (advisor)
In this doctoral thesis, the design of algorithms enabling the implementation of a fully automatic system for vertebrae segmentation in 3D computed tomography (CT) image data of possibly incomplete spines, in patients with bone metastases and vertebral compressions is presented. The proposed algorithm consists of several fundamental problems: spine detection and its axis determination, individual vertebra localization and identification (labeling), and finally, precise segmentation of vertebrae. The detection of the spine, specifically identifying its ends, and determining the course of the spinal canal, combines several advanced methods, including deep learning-based approaches. A novel growing circle method has been designed for tracing the spinal cord canal. Further, the innovative spatially variant filtering of brightness profiles along the spine axis leading to intervertebral disc localization has been proposed and implemented. The discs thus obtained are subsequently identified via comparing the tested vertebrae and model of vertebrae provided by a machine-learning process and optimized by dynamic programming. The final vertebrae segmentation is provided by the deformation of the complete-spine intensity model, utilizing a proposed multilevel registration technique. The complete proposed algorithm has been validated on testing databases, including also publicly available datasets. This way, it has been proven that the newly proposed algorithms provide results at least comparable to other author’s algorithms, and in some cases, even better. The main strengths of the algorithms lie in high reliability of the results and in the robustness to even strongly distorted vertebrae of oncological patients and to the occurrence of artifacts in data; moreover, they are capable of identifying the vertebra labels even in incomplete spinal CT scans. The strength is also in the complete automation of the processing and in its relatively low computational complexity enabling implementation on standard PC hardware. The system for fully automatic localization and labeling of distorted vertebrae in possibly incomplete spinal CT data is presented in this doctoral thesis. The design of algorithms enabling the implementation utilizes several novel approaches, which were presented at international conferences and published in the journal Jakubicek et al. (2020). Based on the results of the experimental validation, the proposed algorithms seem to be routinely usable and capable of providing fully acceptable input data (identified and precisely segmented vertebrae) as needed in the subsequent automatic spine bone lesion analysis.

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