Prolonging the time of fluorescence for Cy3-alkyne and Cy5-alkyne using ROXS

• In Event: Chemistry & Biochemistry Oral Session 3 & Section Meeting

Abstract

Fluorescent dyes, such as Cy3-Alkyne and Cy5-Alkyne, are commonly used in a variety of biological techniques, such as fluorescent tagging and single molecule spectroscopy. The fluorescence signal, of these dyes, decay over time when the fluorophore is repeatedly excited, causing the dye to undergo a permanent loss of fluorescence. Photoprotection systems, such as the reducing and oxidizing system (ROXS), can be used to prevent the fluorophore from photobleaching. ROXS uses the methyl viologen and ascorbic acid redox pair to provide a faster pathway for the electron to travel from the excited triplet state back to the ground state, limiting the exposure time of the highly reactive triplet state of fluorophores. In conjunction with a bulk fluorescence screening process, varying ratios of ROXS to Cy3-Alkyne and Cy5-Alkyne were tested to determine the optimum concentration ratio of photoprotection system to fluorophores in order to maximize the fluorescence time. These results allow for the determination of optimal fluorescence performance, and provide a method for which other photoprotection systems can be rapidly screened and vetted.

Authors

• Mauro A. Garcia, Southwestern University

• Ian Orantes-Orellana, Southwestern University

• Lauren Gillespie, Southwestern University

• Dr. David Cooper, Southwestern University