An optofluidic antenna for enhancing the sensitivity of single-emitter measurements

Luis Morales-Inostroza, Julian Folz, Ralf Kühnemuth, Suren Felekyan, Franz-Ferdinand Wieser, Claus A. M. Seidel (), Stephan Götzinger and Vahid Sandoghdar ()
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Luis Morales-Inostroza: Max Planck Institute for the Science of Light
Julian Folz: Heinrich Heine University Düsseldorf
Ralf Kühnemuth: Heinrich Heine University Düsseldorf
Suren Felekyan: Heinrich Heine University Düsseldorf
Franz-Ferdinand Wieser: Max Planck Institute for the Science of Light
Claus A. M. Seidel: Heinrich Heine University Düsseldorf
Stephan Götzinger: Max Planck Institute for the Science of Light
Vahid Sandoghdar: Max Planck Institute for the Science of Light

Nature Communications, 2024, vol. 15, issue 1, 1-10

Abstract: Abstract Many single-molecule investigations are performed in fluidic environments, for example, to avoid unwanted consequences of contact with surfaces. Diffusion of molecules in this arrangement limits the observation time and the number of collected photons, thus, compromising studies of processes with fast or slow dynamics. Here, we introduce a planar optofluidic antenna (OFA), which enhances the fluorescence signal from molecules by about 5 times per passage, leads to about 7-fold more frequent returns to the observation volume, and significantly lengthens the diffusion time within one passage. We use single-molecule multi-parameter fluorescence detection (sm-MFD), fluorescence correlation spectroscopy (FCS) and Förster resonance energy transfer (FRET) measurements to characterize our OFAs. The antenna advantages are showcased by examining both the slow (ms) and fast (50 μs) dynamics of DNA four-way (Holliday) junctions with real-time resolution. The FRET trajectories provide evidence for the absence of an intermediate conformational state and introduce an upper bound for its lifetime. The ease of implementation and compatibility with various microscopy modalities make OFAs broadly applicable to a diverse range of studies.

Date: 2024
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DOI: 10.1038/s41467-024-46730-w

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