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Microscopy with spatial filtering for monitoring subcelluar morphology
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Microscopy with spatial filtering for monitoring subcelluar morphology
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ETD_2163
Identifier
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http://hdl.rutgers.edu/1782.2/rucore10001600001.ETD.000051936
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eng
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theses
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Biomedical Engineering
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Topic
Microscopy
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Topic
Cells--Morphology
Abstract
Dynamic alteration in organelle morphology is an important indicator of cellular function and many efforts have been made to monitor the subcellular morphology. Optical scatter imaging (OSI), which combines light scattering spectroscopy with microscopic imaging, was developed to non-invasively track real-time changes in particle morphology in situ. Using a variable diameter iris as a Fourier spatial filter, the technique consisted of collecting images that encoded the intensity ratio of wide-to-narrow angle scatter (OSIR, optical scatter imaging ratio) at each pixel in the full field of view. For spherical particles, the OSIR was shown to decrease monotonically with diameter. In living cells, we reported this technique is able to detect mitochondrial morphological alterations, which were mediated by the Bcl-xL transmembrane domain, but could not be observed by fluorescence or DIC images. However, the initial design was based on Mie theory of scattering by spheres, and hence only adequate for measuring spherical particles. This limits the applicability of OSI to cellular functional studies involving organelles, which are naturally non-spherical. In this project, we aim to enhance the current capability of the existing optical scatter microscope to assess size and shape information for both spherical and non-spherical particles, and eventually apply this technique for monitoring and quantifying subcellular morphology within living cells. To reach this goal, we developed an improved system, in which the variable diameter iris is replaced with a digital micromirror device and adopted the concept of Gabor filtering to extend our assessment of morphology to the characterization of particle shape and orientation. Using bacteria and polystyrene spheres, we show how this system can be used to assess particle aspect ratio even when imaged at low resolution. We also show the feasibility of detecting alterations in organelle aspect ratio in situ within living cells. This improved OSI system could be further developed to automate morphological quantification and sorting of non-spherical particles in situ. In the future, we plan to pursue the assessment of morphology information for particles of arbitrary shape and seek to correlate gene expression and cell function with subcellular morphology, which may help in disease diagnosis and drug screening.
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electronic resource
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xi, 112 p. : ill.
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Ph.D.
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Includes bibliographical references (p. 106-111)
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by Jing-Yi Zheng
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Zheng
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Jing-Yi
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Jing-Yi Zheng
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Nada
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Nada Boustany
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Cai
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Li
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Li Cai
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Metaxas
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Dimitris
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Dimitris Metaxas
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Rajadhyaksha
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Milind
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Milind Rajadhyaksha
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White
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Eileen
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Eileen White
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Rutgers University
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Graduate School - New Brunswick
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2009
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2009-10
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xx
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Rutgers University Electronic Theses and Dissertations
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ETD
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Graduate School - New Brunswick Electronic Theses and Dissertations
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rucore19991600001
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doi:10.7282/T3GB2460
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ETD doctoral
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The author owns the copyright to this work
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Zheng
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Jing-Yi
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Jing-Yi Zheng
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Rutgers University. Graduate School - New Brunswick
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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.
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