Multiplexed Fast-Scan Cyclic Voltammetry and Fluorescence Microscopy for Simultaneous Detection of Multiple Neurotransmitters 64 views

This thesis presents a multiplexed method for the simultaneous detection of electroactive and nonelectroactive neurotransmitters through fast-scan cyclic voltammetry (FSCV) and fluorescence microscopy. Chapter 1 presents background information on the two neurotransmitters covered by this thesis: dopamine and glutamate; it also covers relevant techniques to detect either neurotransmitter while providing the motivation for multiplexing FSCV and fluorescence microscopy for simultaneous detection. Chapter 2 explores photochemical interactions between the excitation light used for widefield microscopy and carbon-fiber microelectrodes (CFMEs). Under blue-light exposure, photocurrents are produced which affect FSCV background currents and enhance dopamine detection. Chapter 3 validates multiplexed FSCV and fluorescence microscopy by simultaneously detecting the same neurotransmitter: dopamine. Using two types of genetically-encoded fluorescent sensors, comparisons are drawn between the two techniques and guidance for future experiments is presented. Chapter 4 uses multiplexed FSCV and fluorescence microscopy to examine glutamatergic neuromodulation of dopamine in the mouse nucleus accumbens. The genetically-encoded sensor iGluSnFR was used to detect glutamate, revealing that increasing glutamate concentration inhibits dopamine release. This relationship was determined to be partially mediated through AMPA and Kainate ionotropic receptors. Chapter 5 summarizes the conclusions of the three previous chapters and discusses future directions for biological experiments and technique development.

PHD (Doctor of Philosophy)

Fluorescence Microscopy; Neurotransmitters; Voltammetry

English

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2026-04-15