Single-Molecule Detection

doi:10.1007/978-0-387-46312-4_23

Principles of Fluorescence Spectroscopy 2006

DOI: 10.1007/978-0-387-46312-4_23

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Single-Molecule Detection

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Cited by 2 publications

References 153 publications

(119 reference statements)

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Faster Convergence of Diffusion Anisotropy Detection by Three-Step Relation of Single-Particle Trajectory

et al. 2016

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We focus on the issue of limited number of samples in the single particle tracking (SPT) when trying to extract the diffusion anisotropy that originates from the particle asymmetry. We propose a novel evaluation technique of SPT making use of the relation of the consecutive three steps. More specifically, the trend of the angle comprised of the three positions and the displacements are plotted on a scatter diagram. The particle anisotropy dependence of the shape of the scatter diagram is examined through the data from the standard numerical model of anisotropic two-dimensional Brownian motion. Comparison with the existing method reveals the faster convergence in the evaluation. In particular, our proposed method realizes the detection of diffusion anisotropy under the conditions of not only less number of data but also larger time steps. This is of practical importance not only when the abundant data is hard to achieve but also when the rotational diffusion is fast compared to the frame rate of the camera equipment, which tends to be more common for smaller particles or molecules of interest.

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Faster Convergence of Diffusion Anisotropy Detection by Three-Step Relation of Single-Particle Trajectory

et al. 2016

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Fractal-like Silver Aggregates Enhance the Brightness and Stability of Single-Molecule Fluorescence

et al. 2011

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This paper describes a method to significantly enhance single-molecule fluorescence detection in confocal microscopy, demonstrating that fractal-like silver structures significantly improve dye stability and brightness. The experiments compared two immunoassay models based on the deposition of rabbit IgG on silver structures. The experiments were performed with the fluorophore-labeled protein at low picomolar concentrations. Well-separated bright spots were still easily distinguishable. Under standardized conditions we observed increased photostability and brightness enhancement for the dyes that were immobilized on the surface using a primary antibody. In contrast, when the unlabeled primary antibody was immobilized on the surface and the labeled secondary antibody was placed at a larger distance, we observed only a modest enhancement of fluorescence. Furthermore, based on backscattered reflected light images, it was proven that the observed fluorescence enhancements originate from the areas with deposited silver nanostructures. Fractal-like substrates are relatively easy to prepare. We believe that with their superior performance, they should find wide applications in single-molecule studies in which a longer observation time is required.

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Single-Molecule Detection

Cited by 2 publications

References 153 publications

Faster Convergence of Diffusion Anisotropy Detection by Three-Step Relation of Single-Particle Trajectory

Faster Convergence of Diffusion Anisotropy Detection by Three-Step Relation of Single-Particle Trajectory

Fractal-like Silver Aggregates Enhance the Brightness and Stability of Single-Molecule Fluorescence

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