Light‐Harvesting Nanoparticle Probes for FRET‐Based Detection of Oligonucleotides with Single‐Molecule Sensitivity. Issue 17 (28th February 2020)

• Record Type: Journal Article
• Authors: Melnychuk, Nina; Egloff, Sylvie; Runser, Anne; Reisch, Andreas; Klymchenko, Andrey S.
• Is Part Of: Angewandte Chemie international edition. Volume 59:Issue 17(2020)
• Page Start: 6811
• Page End: 6818
• Publication Date: 2020-02-28
• Subjects: biosensors -- fluorescent probes -- Förster resonance energy transfer -- light-harvesting nanoantennae -- nucleic acids; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201913804
• Languages: English
• ISSNs: 1433-7851

Abstract

Abstract: Controlling the emission of bright luminescent nanoparticles by a single molecular recognition event remains a challenge in the design of ultrasensitive probes for biomolecules. Herein, we developed 20‐nm light‐harvesting nanoantenna particles, built of a tailor‐made hydrophobic charged polymer poly(ethyl methacrylate‐co‐methacrylic acid), encapsulating circa 1000 strongly coupled and highly emissive rhodamine dyes with their bulky counterion. Being 87‐fold brighter than quantum dots QDots 605 in single‐particle microscopy (with 550‐nm excitation), these DNA‐functionalized nanoparticles exhibit over 50 % total FRET efficiency to a single hybridized FRET acceptor, a highly photostable dye (ATTO665), leading to circa 250‐fold signal amplification. The obtained FRET nanoprobes enable single‐molecule detection of short DNA and RNA sequences, encoding a cancer marker (survivin), and imaging single hybridization events by an epi‐fluorescence microscope with ultralow excitation irradiance close to that of ambient sunlight. Abstract : Light‐harvesting nanoprobes are developed based on polymeric nanoparticles encapsulating circa 1000 strongly coupled and highly emissive dye molecules capable of over 50 % FRET efficiency to a single acceptor at the DNA‐functionalized surface. This nanomaterial enables unprecedented detection of single copies of DNA/RNA by using a simple epi‐fluorescence microscope with ultralow excitation power close to that of ambient sunlight.