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We present two novel automated image analysis methods to differentiate centroblast (CB) cells from noncentroblast (non-CB) cells in digital images of HE-stained tissues of follicular lymphoma. CB cells are often confused by similar looking cells within the tissue, therefore a system to help their classification is necessary. Our methods extract the discriminatory features of cells by approximating the intrinsic dimensionality from the subspace spanned by CB and non-CB cells. In the first method, discriminatory features are approximated with the help of singular value decomposition (SVD), whereas in the second method they are extracted using Laplacian Eigenmaps. Five hundred high-power field images were extracted from 17 slides, which are then used to compose a database of 213 CB and 234 non-CB region of interest images. The recall, precision, and overall accuracy rates of the developed methods were measured and compared with existing classification methods. Moreover, the reproducibility of both classification methods was also examined. The average values of the overall accuracy were 99.22% ± 0.75% and 99.07% ± 1.53% for COB and CLEM, respectively. The experimental results demonstrate that both proposed methods provide better classification accuracy of CB/non-CB in comparison with the state of the art methods.

Filtering is a key step in digital image processing and analysis. It is mainly used for amplification or attenuation of some frequencies depending on the nature of the application. Filtering can either be performed in the spatial domain or in a transformed domain. The selection of the filtering method, filtering domain, and the filter parameters are often driven by the properties of the underlying image. This thesis presents three different kinds of biomedical image filtering applications, where the filter parameters are automatically determined from the underlying images. Filtering can be used for image enhancement. We present a robust image dependent filtering method for intensity inhomogeneity correction of biomedical images. In the presented filtering method, the filter parameters are automatically determined from the grey-weighted distance transform of the magnitude spectrum. An evaluation shows that the filter provides an accurate estimate of intensity inhomogeneity. Filtering can also be used for analysis. The thesis presents a filtering method for heart localization and robust signal detection from video recordings of rat embryos. It presents a strategy to decouple motion artifacts produced by the non-rigid embryonic boundary from the heart. The method also filters out noise and the trend term with the help of empirical mode decomposition. Again, all the filter parameters are determined automatically based on the underlying signal. Transforming the geometry of one image to fit that of another one, so called image registration, can be seen as a filtering operation of the image geometry. To assess the progression of eye disorder, registration between temporal images is often required to determine the movement and development of the blood vessels in the eye. We present a robust method for retinal image registration. The method is based on particle swarm optimization, where the swarm searches for optimal registration parameters based on the direction of its cognitive and social components. An evaluation of the proposed method shows that the method is less susceptible to becoming trapped in local minima than previous methods. With these thesis contributions, we have augmented the filter toolbox for image analysis with methods that adjust to the data at hand.