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Omics of Malignant Cells and Tissues Using Spontaneous and Stimulated Raman Spectroscopy

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Title:Omics of Malignant Cells and Tissues Using Spontaneous and Stimulated Raman Spectroscopy
Authors:Oda, Robert Westley
Contributors:Shiramizu, Bruce (advisor)
Molecular Biosciences and Bioengineering (department)
Keywords:Medical imaging
Anal Dysplasia
show 3 moreNon-Hodgkin Lymphoma
Raman Spectroscopy
Stimulated Raman Scattering
show less
Date Issued:2020
Publisher:University of Hawai'i at Manoa
Abstract:Cancer remains a global and pressing issue in medicine. With the need for timely access to care and treatment, timely and efficient cancer diagnosis and detection is necessary. Recently, Raman spectroscopy has emerged as a potential tool in biomedical sciences. As a relatively new technique to assess biological specimens, Raman spectroscopy has been applied to analyze malignant cells and tissues. It is a laser-based tool that characterizes molecular and chemical composition through inelastic scattering of photons. As light passes through chemical bonds, photons interacting with chemical bonds can pass on energy to photons and create a shift in energy.
Traditional Raman scattering, or spontaneous Raman spectroscopy, is a potentially attractive tool for cancer diagnosis as it is label-free and non-destructive for cells. This provides the unique opportunity for faster determination of disease while preserving samples from potentially destructive chemical processes required for pathological determination. The Raman spectra consists of three main areas: the fingerprint (600-1800cm-1), silent (1800-2600 cm-1), and high number C-H regions (2800-3000 cm-1). The fingerprint region is of interest because it provides rich biological information. In this region, information such as DNA/RNA bases, amino acid bonds, and metabolite composition can be observed.
Stimulated Raman spectroscopy (SRS) is an improvement on the classical approach to Raman spectroscopy. In contrast with spontaneous Raman spectroscopy which uses one laser, SRS uses two laser sources: a pump and Stokes pulses. When the difference in pump and Stokes pulses (the Raman shift) equals the vibrational resonance of a certain chemical bond, the signal is amplified by stimulated emission. This allows for stronger, faster imaging of biological materials than spontaneous Raman. Thus, it is possible to visualize organelles of the cell to discriminate between structures in real time and label-free.
This project provides a comprehensive overview of the applications of Raman spectroscopy in cancer diagnosis. The aims are to analyze both cells and tissue in different disease states using spontaneous and stimulated Raman. While spontaneous Raman spectroscopy gives us important insight into the fingerprint region of biological samples, SRS will complement by providing faster acquisition of spectral data to image and expose cellular interactions within the cell. Analysis of cancerous cells and tissue by spontaneous Raman spectroscopy revealed differences between normal and malignant cells and tissue (p < 0.001). Spontaneous Raman spectroscopy was also utilized in a clinical setting and was able to discriminate between normal and cancerous anal tissue in HIV-serodiscordant couples (p < 0.001). In addition, in anal biopsy samples that have the same diagnosis, there were differences in Raman spectra that could be explained by a potential viral effect. And finally, SRS was able to classify cancerous cells within a mixed population of healthy PBMC and cancerous B-cells with a specificity and sensitivity of 80% and 81.2%. Overall, the project set out to show the effectiveness of Raman spectroscopy as a useful tool in diagnostic and advanced omics applications. Results from this project open the door for new and exciting applications of Raman spectroscopy and further work in this field could lead to new avenues of diagnosis and could advance our understanding of the progression of cancer.
Pages/Duration:140 pages
Rights:All UHM dissertations and theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission from the copyright owner.
Appears in Collections: Ph.D. - Molecular Biosciences and Bioengineering

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