Programmed Control of Fluorescence Blinking Patterns based on Electron Transfer in DNA

Abstract

Fluorescence imaging uses changes in the fluorescence intensity and emission wavelength to analyze multiple targets simultaneously. To increase the number of targets that can be identified simultaneously, fluorescence blinking can be used as an additional parameter. To understand and eventually control blinking, we used DNA as a platform to elucidate the processes of electron transfer (ET) leading to blinking, down to the rate constants. With a fixed ET distance, various blinking patterns were observed depending on the DNA sequence between the donor and acceptor units of the DNA platform. The blinking pattern was successfully described with a combination of ET rate constants. Therefore, molecules with various blinking patterns can be developed by tuning ET. It is expected that the number of targets that can be analyzed simultaneously will increase by the power of the number of blinking patterns.

Fluorescence Bar-Coding and Flowmetry Based on Dark State Transitions in Fluorescence Emitters

“…Monitoring the blinking such states generate, by single-molecule, FCS, or transient state (TRAST) spectroscopy, can thus form the basis for microenvironmental sensing applications. More recent examples include FCS-monitored quantum dot (QD) blinking to probe QD dimerization following mRNA hybridization single-molecule blinking measurements of QDs and organic fluorophores as a basis for multiplexing or of different fluorophore-labeled DNA molecules, designed to have differences in their electron transfer rates . By transient state (TRAST) monitoring, such states can be followed by a relatively simple approach, from how the time-averaged fluorescence intensity from the fluorophores varies with the modulation of the laser excitation intensity. ,− Since TRAST, in contrast to FCS, does not rely on single-molecule detection conditions or a high time resolution, it can be applied on a broader range of samples.…”