Date of Graduation
Bachelor of Science
Chemistry & Biochemistry
Committee Member/Second Reader
Committee Member/Third Reader
Committee Member/Fourth Reader
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.
two-photon excitation, NADH, FAD, fluorescence, redox ratio, FLIM
Raley, Robin L., "Autofluorescence to Study the Effects of Acid Concentration on Cellular Metabolism in Vitro" (2019). Chemistry & Biochemistry Undergraduate Honors Theses. 24.
Available for download on Saturday, April 30, 2022