Date of Graduation

5-2019

Document Type

Thesis

Degree Name

Bachelor of Science

Degree Level

Undergraduate

Department

Chemistry & Biochemistry

Advisor/Mentor

Quinn, Kyle

Committee Member/Reader

Koeppe, Roger

Committee Member/Second Reader

Chen, Jingyi

Committee Member/Third Reader

Harrington, Phil

Committee Member/Fourth Reader

Quinn, Kyle

Abstract

Ultraviolet (UV) radiation-induced sunburns and their accompanying afflictions are a growing public health concern in the United States. There is a need for techniques that can accurately and non-invasively characterize the physiology of sunburned skin tissue directly after UV-damage and applying a topical skin treatment to relieve pain and promote healing. Two-photon excited fluorescence (TPEF) microscopy and fluorescence lifetime imaging (FLIM) can be used to investigate metabolic processes in live cells through endogenous fluorescence of the cofactors, NADH and FAD. These methods employ the optical redox ratio of FAD/(NADH+FAD), mean NADH lifetime, and the separation of the free and bound NADH lifetime components to quantify glycolytic activity relative to oxidative phosphorylation. In this study, I probe the metabolism dynamics of mouse fibroblasts when treated with media containing increasing concentrations of an acid* as a first step to evaluate the effects of topical application of this acid to skin as a treatment for sunburns. I am able to show that significant increases in the redox ratio due to increasing Acid X treatment, in vitro, suggest that Acid X may shift metabolism in cells in a concentration-dependent manner. The application of methods here demonstrates that autofluorescence optical imaging can serve as a quantitative measure of cellular metabolism changes under the application of Acid X.

*Note: This acid will be referred to as “Acid X” throughout this thesis.

Keywords

two-photon excitation; NADH; FAD; fluorescence; redox ratio; FLIM

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