Date of Award

Spring 5-10-2025

Level of Access Assigned by Author

Open-Access Thesis

Degree Name

Master of Science (MS)

Department

Chemistry

First Committee Advisor

Karissa Tilbury

Second Committee Member

Matthew Brichacek

Third Committee Member

Michael Mason

Abstract

Two-photon (2P) microscopy is a powerful biomedical imaging technique that has vast implications for investigation of cancerous tumors. The utilization of 2P microscopy to quantify cellular processes is unique in that visualization of tissues surrounding cellular structures can be achieved at sub-micrometer resolution while avoiding exogeneous labeling. The work performed in this thesis describes all the necessary principles and practical knowledge needed to integrate with biological imaging data generated from a 2P microscope. Our 2P microscope was utilized in imaging spheroid models of the tumor microenvironment for the purpose of understanding the changes in the collagen matrix surrounding breast cancer cells. The forward (F) to backward (B) ratio of the second harmonic generation (SHG) scattering from the collagen fibers reveals distinct biological changes in the tumor microenvironment of breast cancer cells. The fundamental photo-physics is explained in parallel to the practical methods of optical alignment, imaging calibration, and laser maintenance to facilitate the interpretation of model biological data. The knowledge contained within this thesis works to produce a heightened understanding of the practical implications of the SHG process to yield forward progress in our understanding of cancer biology.

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