Traditional laser scanning systems, such as those used for microscopy, typically image objects of finite thickness. If the depth-of-focus of such systems is low, as is the case when a simple clear pupil is used, the object must be very thin or the image will be distorted. Several methods have been developed to deal with this problem. A microscope with a thin annular pupil has a very high depth-of-focus and can image the entire thickness of a sample, but most of the laser light is blocked, and the image shows poor contrast and high noise. In confocal laser microscopy, the depth-of-focus problem is eliminated by using a small aperture to discard information from all but one thin plane of the sample. However, such a system requires scanning passes at many different depths to yield an image of the entire thickness of the sample, which is a time-consuming process and is highly sensitive to registration errors. In this thesis, a novel type of scanning system is considered. The sample is simultaneously scanned with a combination of two Gaussian laser beams of different widths and slightly different temporal frequencies. Information from scanning with the two beams is recorded with a photodetector, separated electronically, and processed to form an image. This image is similar to one formed by a system using a difference-of-Gaussians pupil, except no light has been blocked or wasted. Also, the entire sample can be scanned in one pass. The depth-of-focus characteristics of this synthesized difference-of-Gaussians pupil are examined and compared with those of well-known / Master of Science

Axicons have been proposed for imaging applications since they offer the advantage of extended depth of field (DOF). This enhanced DOF comes at the cost of degraded image quality. Image processing has been proposed to improve the image quality. Initial efforts were focused on the use of an axicon in a borescope thereby extending depth of focus and eliminating the need for a focusing mechanism. Though promising, it is clear that image processing would lead to improved image quality. This would also eliminate the need, in certain applications, for a fiber optic imaging bundle as many modern day video borescopes use an imaging sensor coupled directly to the front end optics. In the present work, three types of refractive axicons are examined: a linear axicon, a logarithmic axicon and a Fresnel axicon. The linear axicon offers the advantage of simplicity and a significant amount of scientific literature including the application of image restoration techniques. The Fresnel axicon has the advantage of compactness and potential low cost of production. As no physical prior examples of the Fresnel axicons were available for experimentation until recently, very little literature exists. The logarithmic axicon has the advantage of nearly constant longitudinal intensity distribution and an aspheric design producing superior pre-processed images over the aforementioned elements. Point Spread Functions (PSFs) for each of these axicons have been measured. These PSFs form the basis for the design of digital image restoration filters. The performance of these three optical elements and a number of restoration techniques are demonstrated and compared.

Dans le circuit de la greffe de cornée, l'endothélium de chaque greffon est observé en microscopie optique conventionnelle afin de vérifier que sa densité cellulaire est suffisante pour maintenir une bonne transparence après l'opération. Les greffons étant conservés dans un milieu spécifique, ils sont imprégnés de liquide et présentent donc des plis qui perturbent l'observation et le comptage des cellules. Ce problème pratique est à l'origine d’une étude théorique sur les concepts de profondeur de champ étendue et de shape-from-focus. A partir d'une séquence d'images acquise par sectionnement optique, les informations les plus nettes permettent d'une part d'accéder à la topographie de la surface observée et d'autre part de restaurer l'image de sa texture. Une reconstruction surfacique 3-D est alors obtenue en projetant la texture sur la topographie. Cette thèse considère essentiellement l’étape fondamentale de mesure de netteté du processus de reconstruction. Des nouvelles mesures génériques offrant une haute sensibilité à la netteté sont introduites. De par une stratégie 3-D originale au travers de la séquence d'images, une autre mesure très robuste au bruit est proposée. Toutes ces mesures sont testées sur des données simulées puis diverses acquisitions réelles en microscopie optique conventionnelle et comparées aux méthodes de la littérature. Par ailleurs, la mesure 3-D améliore nettement les reconstructions d'endothéliums cornéens à partir de leurs acquisitions particulièrement perturbées (inversions de contraste). Un processus itératif complet de reconstruction 3-D d’endothéliums cornéens est finalement décrit, aboutissant à des résultats solides et exploitables. / In the cornea transplant process, each graft endothelium is observed by conventional optical microscopy to check that its cell density is sufficient to maintain a proper transparency after the transplantation. The grafts are stored in a specific preservation medium, they are thus impregnated with fluid and therefore exhibit folds which make cell observation and counting difficult. This practical issue led to the following theoretical study about the so-called concepts: extended-depth-of-field and shape-from-focus. Throughout a sequence of images acquired by optical sectioning, the in-focus information allows on the one hand to recover the topography of the observed surface and on the other hand to restore the image of its texture. A 3-D reconstruction is then obtained by mapping the texture onto the topography. This thesis basically considers the fundamental step of the reconstruction process that is the focus measurement. New generic focus measurements exhibiting high sharpness sensitivity are introduced. Another one offering high noise robustness is proposed, due to an original 3-D strategy through the image sequence, unlike traditional methods that operate in 2-D. All of them are tested on simulated data and various real acquisitions, and compared to the state-of-the-art methods. Furthermore, the aforementioned 3-D focus measurement clearly improves the 3-D surface reconstructions of the corneal endotheliums from their particularly disturbed acquisitions (contrast reversals). A complete iterative process of 3-D reconstruction of the corneal endothelial surfaces is finally described, resulting in solid results that can already be transferred to cornea banks.