High-resolution microendoscopy (HRME) uses epi-fluorescence imaging with a coherent fiber-optic bundle to enable in vivo examination of cellular morphology. While the HRME platform has recently gained popularity as a simple alternative to confocal endomicroscopy, the axial response of HRME in thick, scattering tissue has yet to be described quantitatively. This is important because when analyzing images collected by HRME, out-of-focus light may affect the accuracy of quantitative parameters such as nuclear-to-cytoplasm ratio, which has been proposed as a diagnostic indicator of dysplasia or cancer. In the first part of this thesis, the imaging properties of the HRME system are investigated using phantoms simulating scattering tissue with fluorescently labeled nuclei. HRME images defocused (deep) objects with apparent diameters and intensity levels that are in agreement with a simple geometric model. Out-of-focus nuclei contribute a relatively low, uniform background level to images which neither leads to the erroneous appearance of large nuclei from deep layers, nor prevents accurate imaging of superficial nuclei with high contrast. Proflavine has been used as a fluorescent contrast agent for HRME imaging, brightly labeling nuclei without staining the surrounding cytoplasm or organelles. If a non-specific fluorophore such as fluorescein is used, the removal of out-of-focus (OOF) light would be desirable. Optical sectioning is a property of some imaging systems, whereby OOF signal light is removed or rejected from the image. Optical sectioning is commonly associated with confocal microscopy and non-linear imaging methods such as multi-photon microscopy. In contrast, structured illumination (SI) is an imaging modality which is capable of providing optical sectioning without requiring raster scanning of a tightly focused laser beam. SI was integrated with the HRME platform and demonstrated to be an effective method of suppressing OOF signals, with comparable results to confocal. Theory, example images, programming considerations, and methods to minimize artifacts specific to SI are considered.
Subject (authority = RUETD)
Topic
Biomedical Engineering
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TitleInfo
Title
Rutgers University Electronic Theses and Dissertations
Identifier (type = RULIB)
ETD
Identifier
ETD_5270
PhysicalDescription
Form (authority = gmd)
electronic resource
InternetMediaType
application/pdf
InternetMediaType
text/xml
Extent
viii, 51 p. : ill.
Note (type = degree)
M.S.
Note (type = bibliography)
Includes bibliographical references
Note (type = statement of responsibility)
by Michael Harten Koucky
Subject (authority = ETD-LCSH)
Topic
Endoscopy
Subject (authority = ETD-LCSH)
Topic
Imaging systems in medicine
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TitleInfo
Title
Graduate School - New Brunswick Electronic Theses and Dissertations
Identifier (type = local)
rucore19991600001
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PhysicalLocation (authority = marcorg); (displayLabel = Rutgers, The State University of New Jersey)
Rutgers University. Graduate School - New Brunswick
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License
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Author Agreement License
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I hereby grant to the Rutgers University Libraries and to my school the non-exclusive right to archive, reproduce and distribute my thesis or dissertation, in whole or in part, and/or my abstract, in whole or in part, in and from an electronic format, subject to the release date subsequently stipulated in this submittal form and approved by my school. I represent and stipulate that the thesis or dissertation and its abstract are my original work, that they do not infringe or violate any rights of others, and that I make these grants as the sole owner of the rights to my thesis or dissertation and its abstract. I represent that I have obtained written permissions, when necessary, from the owner(s) of each third party copyrighted matter to be included in my thesis or dissertation and will supply copies of such upon request by my school. I acknowledge that RU ETD and my school will not distribute my thesis or dissertation or its abstract if, in their reasonable judgment, they believe all such rights have not been secured. I acknowledge that I retain ownership rights to the copyright of my work. I also retain the right to use all or part of this thesis or dissertation in future works, such as articles or books.