Date of Award

1-1-2022

Language

English

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

College/School/Department

Department of Physics

Content Description

1 online resource (vi, 86 pages) : illustrations (some color)

Dissertation/Thesis Chair

Alexander Khmaladze

Committee Members

Jonathan C Petruccelli, William A Lanford, Margarida M Barroso, Supriya D Mahajan, Alexander Khmaladze

Keywords

Digital holographic microscopy, Hyperspectral imaging, Iron bound transfferrin, Quantitative phase imaging, Raman spectroscopy, Transport of intensity equation, Imaging systems in medicine, Breast

Subject Categories

Optics | Physics

Abstract

This thesis presents the application of Raman spectroscopy, digital holographic microscope (DHM) and transport intensity equation (TIE) in imaging biological samples. Raman spectroscopy is a non-destructive technique that can provide chemical structure, concentration, temperature, and molecular interactions. On the other hand, DHM and TIE provide physical measurements such as height, width, area and volume.We applied Raman spectroscopy to study iron-bound transferrin (Tf) in intact human breast cancer cells. Iron is an essential element required for human life and is highly regulated in the body. As the exact mechanisms of iron-bound Tf in cells are not well known, we developed a Raman micro-spectroscopy technique that allows detection of iron-bound transferrin (Tf) inside intact human breast cancer cells. Specifically, we applied hyperspectral Raman imaging to visualize the spatial distribution of Tf-bound iron in human breast cancer T47D and MDAMB231 cells internalized with iron-loaded and iron-depleted Tf. This methodology provides a unique way to determine the amount of iron-bound Tf under different Tf internalization times. Employing Singular Value Decomposition (SVD) analysis to spectra allows us to visualize and identify the important peaks that correspond to iron content in iron bound Tf (holo-Tf). Importantly, Raman micro-spectroscopy can be used to follow iron release from Tf in breast cancer cells to further our understanding of iron metabolism during cancer progression. We combined DHM and TIE techniques to validate the TIE accuracy in imaging biological cells. DHM is an interferometric technique which requires a high level of optics alignment to build a working setup. DHM setup is also costly and often suffers from speckle noise that comes from coherent laser source. Introducing TIE coupled with DHM was meaningful as TIE setup is much easier to align and cost less in equipment compared to DHM. We tested DHM and TIE configuration in imaging human cheek cells, and then improved the system by implementing an extra camera to achieve simultaneous measurement of C6 glial cells after hydrogen peroxide (H2O2) treatment.

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