Controlled reduction of photobleaching in DNA origami gold nanoparticle hybrids

The amount of information obtainable from a fluorescence-based measurement is limited by photobleaching: Irreversible photochemical reactions either render the molecules nonfluorescent or shift their absorption and/or emission spectra outside the working range. Photobleaching is evidenced as a decrease of fluorescence intensity with time, or in the case of single molecule measurements, as an abrupt, single-step interruption of the fluorescence emission that determines the end of the experiment. Reducing photobleaching is central for improving fluorescence (functional) imaging, single molecule tracking, and fluorescence-based biosensors and assays. In this single molecule study, we use DNA self-assembly to produce hybrid nanostructures containing individual fluorophores and gold nanoparticles at a controlled separation distance of 8.5 nm. By changing the nanoparticles? size we are able to systematically increase the mean number of photons emitted by the fluorophores before photobleaching.Fil: Pellegrotti, Jesica Vanesa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Centro de Investigaciones en Bionanociencias "Elizabeth Jares Erijman"; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Física; ArgentinaFil: Acuña, Guillermo. Technische Universität Braunschweig. Institute for Physical and Theoretical Chemistry. NanoBioSciences Group; AlemaniaFil: Puchkova, Anastasiya. Technische Universität Braunschweig. Institute for Physical and Theoretical Chemistry. NanoBioSciences Group; AlemaniaFil: Holzmeister, Phil. Technische Universität Braunschweig. Institute for Physical and Theoretical Chemistry. NanoBioSciences Group; AlemaniaFil: Gietl, Andreas. Technische Universität Braunschweig. Institute for Physical and Theoretical Chemistry. NanoBioSciences Group; AlemaniaFil: Lalkens, Birka. Technische Universität Braunschweig. Institute for Physical and Theoretical Chemistry. NanoBioSciences Group; AlemaniaFil: Stefani, Fernando Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Centro de Investigaciones en Bionanociencias "Elizabeth Jares Erijman"; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Física; ArgentinaFil: Tinnefeld, Philip. Technische Universität Braunschweig. Institute for Physical and Theoretical Chemistry. NanoBioSciences Group; Alemani

DNA Origami Nanoantennas with over 5000-fold Fluorescence Enhancement and Single-Molecule Detection at 25 μM

Optical nanoantennas are known to focus freely propagating light and reversely to mediate the emission of a light source located at the nanoantenna hotspot. These effects were previously exploited for fluorescence enhancement and single-molecule detection at elevated concentrations. We present a new generation of self-assembled DNA origami based optical nanoantennas with improved robustness, reduced interparticle distance, and optimized quantum-yield improvement to achieve more than 5000-fold fluorescence enhancement and single-molecule detection at 25 μM background fluorophore concentration. Besides outperforming lithographic optical antennas, DNA origami nanoantennas are additionally capable of incorporating single emitters or biomolecular assays at the antenna hotspot

Controlled Reduction of Photobleaching in DNA Origami–Gold Nanoparticle Hybrids

The amount of information obtainable from a fluorescence-based measurement is limited by photobleaching: Irreversible photochemical reactions either render the molecules nonfluorescent or shift their absorption and/or emission spectra outside the working range. Photobleaching is evidenced as a decrease of fluorescence intensity with time, or in the case of single molecule measurements, as an abrupt, single-step interruption of the fluorescence emission that determines the end of the experiment. Reducing photobleaching is central for improving fluorescence (functional) imaging, single molecule tracking, and fluorescence-based biosensors and assays. In this single molecule study, we use DNA self-assembly to produce hybrid nanostructures containing individual fluorophores and gold nanoparticles at a controlled separation distance of 8.5 nm. By changing the nanoparticles’ size we are able to systematically increase the mean number of photons emitted by the fluorophores before photobleaching

Can physiological sleepiness underlie consciously perceived sleepiness assessed with the Epworth sleepiness scale?

Methods of evaluation of substates of sleep and sleepiness differ in, at least, two respects. Although sleepiness has not been separated from other wake and sleep substates using yes-or-no criteria for sleep scoring, it would be consciously perceived and, therefore, assessed with a questionary. However, such subjective method was challenged by the finding suggesting a disconnect between two most widely used subjective and objective indicators of excessive daytime sleepiness (EDS), a score on the Epworth sleepiness scale (ESS) > 10 and a reduced latency to sleep onset (SOL), respectively. We examined whether these two EDS indicators differ in their association with physiological sleepiness, i.e., the polysomnographic indexes of elevated sleep pressure. In the afternoon hours, polysomnographic recordings were obtained throughout 54 50-min and 56 90-min napping attempts of 27 and 28 university students, respectively. Within some but not all 10-min intervals of the 50- or 90-min naps, each EDS indicator was validated against different objective polysomnographic indexes suggesting an association of EDS with elevated sleep pressure. Significant differences in sleep indexes were found between participants with short and longer SOL, but they disappeared right before the appearance of such differences between participants with higher and lower ESS score (usually at the 4th 10-min interval). This mismatch in timing of appearance of significant differences might be a plausible explanation for the lack of significant association between the ESS and SOL. Therefore, the physiologic underpinnings of the ESS can be uncovered despite such a disconnect between these two EDS indicators

Can physiological sleepiness underlie consciously perceived sleepiness assessed with the Epworth sleepiness scale?

Abstract Methods of evaluation of substates of sleep and sleepiness differ in, at least, two respects. Although sleepiness has not been separated from other wake and sleep substates using yes-or-no criteria for sleep scoring, it would be consciously perceived and, therefore, assessed with a questionary. However, such subjective method was challenged by the finding suggesting a disconnect between two most widely used subjective and objective indicators of excessive daytime sleepiness (EDS), a score on the Epworth sleepiness scale (ESS) > 10 and a reduced latency to sleep onset (SOL), respectively. We examined whether these two EDS indicators differ in their association with physiological sleepiness, i.e., the polysomnographic indexes of elevated sleep pressure. In the afternoon hours, polysomnographic recordings were obtained throughout 54 50-min and 56 90-min napping attempts of 27 and 28 university students, respectively. Within some but not all 10-min intervals of the 50- or 90-min naps, each EDS indicator was validated against different objective polysomnographic indexes suggesting an association of EDS with elevated sleep pressure. Significant differences in sleep indexes were found between participants with short and longer SOL, but they disappeared right before the appearance of such differences between participants with higher and lower ESS score (usually at the 4th 10-min interval). This mismatch in timing of appearance of significant differences might be a plausible explanation for the lack of significant association between the ESS and SOL. Therefore, the physiologic underpinnings of the ESS can be uncovered despite such a disconnect between these two EDS indicators. Graphical abstrac