Cellular metabolism represents all the series of reactions in a cell that take place in order for the cell to survive. Tissue-like environments simulate a shared network of cells by introducing different forms of turbid media to a cell solution to simulate a real-life environment. Autofluorescence is a method for analyzing metabolic response in biological organisms, which can be quantified in a spectral phasor analysis plot. One way to induce autofluorescence is to use a nitrogen laser system with a staging area for cuvettes. The current nitrogen laser system in the lab is large and non-mobile so an LED system consisting of orthogonal source detector geometry with long pass filtering before a spectrofluorometer was developed in the hopes to make data measurements more mobile. To create tissue-like environments, both hemoglobin and polystyrene microspheres were used in a cell solution containing PBS and yeast cells. Spectral phasor analysis of the hemoglobin showed a down and left shift for the addition of hemoglobin and a down and left shift for the addition of potassium cyanide among both the nitrogen laser and LED systems. Spectral phasor analysis of the polystyrene showed an up and right shift for the addition of polystyrene and a down and left shift for the addition of potassium cyanide among both the nitrogen laser and LED system. We conclude that the designed LED system is a viable source for recreating measurements of induced autofluorescence taken on the nitrogen laser system, but slight discrepancies in the phasor plots show that this project is still a work in progress.
Authors: Blake McClain, Nick Majer
Advisor: Paul Urayama, Physics
Graduate Advisor: Mary McNeill, Physics
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