Fluorophore photostability and saturation in the hotspot of DNA origami nanoantennas
Creators
- 1. Department of Chemistry and Center for NanoScience, Ludwig-Maximilians-Universität München, Butenandtstr. 5-13, 81377 München, Germany
- 2. Department of Chemistry and Center for NanoScience, Ludwig-Maximilians-Universität München, Butenandtstr. 5-13, 81377 München, Germany; Institute for Physical and Theoretical Chemistry—NanoBioScience and Braunschweig Integrated Centre of Systems Biology (BRICS), Technische Universität Braunschweig, Braunschweig, Germany
Description
Fluorescent dyes used for single-molecule spectroscopy can undergo millions of excitation-emission
cycles before photobleaching. Due to the upconcentration of light in a plasmonic hotspot, the
conditions for fluorescent dyes are even more demanding inDNAorigami nanoantennas. Here, we
briefly review the current state of fluorophore stabilization for single-molecule imaging and reveal
additional factors relevant in the context of plasmonic fluorescence enhancement. Weshow that
despite the improved photostability of single-molecule fluorophores byDNAorigami nanoantennas,
their performance in the intense electric fields in plasmonic hotspots is still limited by the underlying
photophysical processes, such as formation of dim states and photoisomerization. These photophysical
processes limit the photon count rates, increase heterogeneity and aggravate quantification of
fluorescence enhancement factors. These factors also reduce the time resolution that can be achieved
in biophysical single-molecule experiments. Finally, we show how the photophysics of aDNAhairpin
assay with a fluorophore-quencher pair can be influenced by plasmonicDNAorigami nanoantennas
leading to implications for their use in fluorescence-based diagnostic assays. Especially, we show that
such assays can produce false positive results by premature photobleaching of the dark quencher. Here we demonstrate the raw data on which our findings based on.
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