Improved dye stability in single-molecule fluorescence experiments

0Citations
Citations of this article
10Readers
Mendeley users who have this article in their library.
Get full text

Abstract

Complex biological systems challenge existing single-molecule methods. In particular, dye stability limits observation time in singlemolecule fluorescence applications. Current approaches to improving dye performance involve the addition of enzymatic oxygen scavenging systems and small molecule additives. We present an enzymatic oxygen scavenging system that improves dye stability in single-molecule experiments. Compared to the currently-employed glucose-oxidase/catalase system, the protocatechuate-3,4-dioxygenase system achieves lower dissolved oxygen concentration and stabilizes single Cy3, Cy5, and Alexa488 fluorophores. Moreover, this system possesses none of the limitations associated with the glucose oxidase/catalase system. We also tested the effects of small molecule additives in this system. Biological reducing agents significantly destabilize the Cy5 fluorophore as a function of reducing potential. In contrast, anti-oxidants stabilize the Cy3 and Alexa488 fluorophores. We recommend use of the protocatechuate-3,4,-dioxygenase system with antioxidant additives, and in the absence of biological reducing agents. This system should have wide application to single-molecule fluorescence experiments. © 2009 Springer Netherlands.

Cite

CITATION STYLE

APA

EcheverrÍa Aitken, C., Marshall, R. A., & Pugi, J. D. (2009). Improved dye stability in single-molecule fluorescence experiments. In NATO Science for Peace and Security Series B: Physics and Biophysics (pp. 83–99). Springer Verlag. https://doi.org/10.1007/978-90-481-2368-1_6

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Save time finding and organizing research with Mendeley

Sign up for free