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Contribution to fluorescence microscopy, 3D thick samples deconvolution and depth-variant PSF

The 3-D fluorescence microscope has become the method of choice in biological sciences for living cells study. However, the data acquired with conventional3-D fluorescence microscopy are not quantitatively significant because of distortions induced by the optical acquisition process. Reliable measurements need the correction of theses distortions. Knowing the instrument impulse response, also known as the PSF, one can consider the backward process of convolution induced by the microscope, known as "deconvolution". However, when the system response is not invariant in the observation field, the classical algorithms can introduce large errors in the results. In this thesis we propose a new approach, which can be easily adapted to any classical deconvolution algorithm, direct or iterative, for bypassing the non-invariance PSF problem, without any modification to the later. Based on the hypothesis that the minimal error in a restored image using non-invariance assumption is located near the used PSF position, the EMMA (Evolutive Merging Masks Algorithm) blends multiple deconvolutions in the invariance assumption using a specific merging mask set. In order to obtain sufficient number of measured PSF at various depths for a better restoration using EMMA (or any other depth-variant deconvolution algorithm) we propose a 3D PSF interpolation algorithm based on the image moments theory using Zernike polynomials as decomposition base. The known PSF are decomposed into Zernike moments set and each moment's variation is fitted into a polynomial function, the resulting functions are then used to interpolate the needed PSF's Zernike moments set to reconstruct the interpolated PSF.

Identiferoai:union.ndltd.org:CCSD/oai:tel.archives-ouvertes.fr:tel-00594247
Date20 December 2010
CreatorsMaalouf, Elie
PublisherUniversité de Haute Alsace - Mulhouse
Source SetsCCSD theses-EN-ligne, France
LanguageEnglish
Detected LanguageEnglish
TypePhD thesis

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