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Single-molecule biosensors employing fluorescence microscopy enable direct counting of molecular targets such as nucleic acids and proteins. Whereas highly specific sandwich assays have been developed using fluorescence probes these assays provide intrinsically low single-molecule fluorescence signals. As a result, these sensors are not compatible with complex biological fluids such as blood serum, and they require research-grade optical microscopies.
I will present our recent work on nanoplasmonic biosensors that exploit plasmon-enhanced fluorescence (PEF) thereby enabling single-molecule biosensing with a strongly enhanced signal-to-noise ratio. The biomarker concentration is quantified by the direct counting of single-molecule interactions, in this case for DNA. The strongly enhanced fluorescence signals enable single-molecule sensing directly in full blood serum with negligible background. Employing fluorescent probes with different emission wavelengths allows for multiplexed biosensing. The bright signals allow for single-molecule sensing in a highly miniaturized optical microscope. This further obviates the need for high numerical aperture objective lenses and paves the way to deployment of the sensors in healthcare and environmental applications.
Peter Zijlstra
"Single-molecule biosensing using plasmon-enhanced fluorescence", Proc. SPIE PC12654, Enhanced Spectroscopies and Nanoimaging 2023, PC1265409 (5 October 2023); https://doi.org/10.1117/12.2677322
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Peter Zijlstra, "Single-molecule biosensing using plasmon-enhanced fluorescence," Proc. SPIE PC12654, Enhanced Spectroscopies and Nanoimaging 2023, PC1265409 (5 October 2023); https://doi.org/10.1117/12.2677322