Single molecule Fluorescence Resonance Energy Transfer (FRET) has become an extremely popular due to its wide range of applications. By using FRET on a confocal setup, the distances between two dyes on a functioning protein can be determined. This can help further studies on protein folding and other bio-conformational process. To calculate the distances between the dyes there are many factors that need to be considered for accurate analysis. In this research, the main focus is accurately determining the fluorescence quantum yield of the dyes used in FRET. This is important because the samples produced for FRET analysis have low concentrations (nanomolar) eliminating the ability to take their absorbances. Therefore, Fluorescence Correlation Spectroscopy (FCS) method with molecular brightness analysis can be developed to determine the fluorescence quantum yield of dye samples in different environment. In this research, the initial steps for this process were performed by collecting FCS data on free Alexa 488 dye using different detergent. To obtain the optimal experimental parameters for FCS analysis on a confocal microscope.

Developing fluorescence correlation spectroscopy (FCS) method for fluorescence quantum yield determination

2019 Spring National Meeting

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