The electrochemical modulation of single molecule fluorescence

Abstract

Recently it has been shown that electrochemistry, instead of using high intensity lasers, can be used to modulate the intensity of emission of fluorophores and even switch fluorophores between their ON and OFF states as required for single molecule localisation microscopy. This modulation of fluorescence does not necessarily correlate with direct oxidation and reduction of the dyes. Questions arise from this unexpected observation related to what is the electrochemistry that occurs, what are the important variables in switching fluorophores electrochemically and what range of dyes can be modulated with electrochemistry. Herein we seek to answer some of these questions. We demonstrate how to effectively modulate the fluorescence intensity of organic dye-labelled cell samples on an indium tin oxide surface using electrochemistry with redox-active mediators present in an oxygen scavenger buffer. We showed the electrochemical fluorescence modulation is sensitive to the applied potential and the excitation laser intensity, indicating the possibility of coupled photochemical and electrochemical reactions occurring. We also compared the electrochemical fluorescence modulation of representative oxazine, rhodamine, and cyanine dyes using ATTO 655, Alexa Fluor 488, and Alexa Fluor 647. Different dyes with distinctly different structural cores show fluorescence modulation to different extents. The electrochemical fluorescence modulation will be applicable in fluorescence imaging techniques as well as biosensing.