C48-P: Distinguishing Between Cellular Respiration and Oxidative Stress Using Autofluorescence Spectroscopy in a Tissue-like Background

My project involves using laser autofluorescence spectroscopy paired with spectral phasor analysis, a biophysical technique, to distinguish metabolic changes in yeast cells. I have specifically focused on measuring changes associated with reduced nicotinamide adenine dinucleotide (NADH) and nicotinamide adenine dinucleotide phosphate (NADPH), which are metabolic cofactors associated with cellular respiration and oxidative stress, respectively. Their nearly identical properties make them difficult to distinguish, especially in the presence of a tissue-like background. In this case we have used 9-cyanoanthrocene (9CA) to mimic the type of background signal found in human tissue. Through experimental data collection and analysis we have been able to see non-linear, and therefore distinguishable, phasor shifts associated with these two co-factors, both with and without the 9CA background signal. This indicates that distinguishing between problems with cellular respiration and oxidative stress is possible using this type of technique. The largest biomedical applications of these results are in disease detection and diagnoses of many metabolically linked diseases, such as various forms of cancer, Alzheimer’s disease, diabetes and heart disease. Going forward, we hope to continue to see these detectable changes with a variety of background signals, chemical concentrations, and eventually move on to taking data in live human tissue.

Author: Taylor Phillips

Faculty Advisor: Paul Urayama, Physics

Graduate Student Advisor: Bibek Dhakal, Physics

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