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Spectrally coded optical nanosectioning (SpecON) with biocompatible metal-dielectric-coated substrates.

ABSTRACT: Fluorescence nanosectioning within a submicron region above an interface is desirable for many disciplines in the life sciences. A drawback, however, to most current approaches is the a priori need to physically scan a sculptured point spread function in the axial dimension, which can be undesirable for optically sensitive or highly dynamic samples. Here we demonstrate a fluorescence imaging approach that can overcome the need for scanning by exploiting the position-dependent emission spectrum of fluorophores above a simple biocompatible nanostructure. To achieve this we have designed a thin metal-dielectric-coated substrate, where the spectral modification to the total measured fluorescence can be used to estimate the axial fluorophore distribution within distances of 10-150 nm above the substrate with an accuracy of up to 5-10 nm. The modeling and feasibility of the approach are verified and successfully applied to elucidate nanoscale adhesion protein and filopodia dynamics in migrating cells. It is likely that the general principle can find broader applications in, for example, single-molecule studies, biosensing, and studying fast dynamic processes.

SUBMITTER: Elsayad K

PROVIDER: S-EPMC3864330 | biostudies-literature | 2013 Dec

REPOSITORIES: biostudies-literature

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Spectrally coded optical nanosectioning (SpecON) with biocompatible metal-dielectric-coated substrates.

Elsayad Kareem K Urich Alexander A Tan Piau Siong PS Nemethova Maria M Small J Victor JV Unterrainer Karl K Heinze Katrin G KG

Proceedings of the National Academy of Sciences of the United States of America 20131125 50

Fluorescence nanosectioning within a submicron region above an interface is desirable for many disciplines in the life sciences. A drawback, however, to most current approaches is the a priori need to physically scan a sculptured point spread function in the axial dimension, which can be undesirable for optically sensitive or highly dynamic samples. Here we demonstrate a fluorescence imaging approach that can overcome the need for scanning by exploiting the position-dependent emission spectrum o ...[more]

PMID: 24277837

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Project description:We describe a hybrid metal-dielectric waveguide structures (MDWs) with numerous potential applications in the biosciences. These structures consist of a thin metal film coated with a dielectric layer. Depending on the thickness of the dielectric layer, the modes can be localized near the metal, within the dielectric, or at the top surface of the dielectric. The optical modes in a metal-dielectric waveguide can have either S (TE) or P (TM) polarization. The dielectric spacer avoids the quenching, which usually occurs for fluorophores within about 5 nm from the metal. Additionally, the resonances display a sharp angular dependence and can exhibit several hundred-fold increases in intensity (E2) at the silica-air interface relative to the incident intensity. Fluorophores placed on top of the silica layer couple efficiently with the metal, resulting in a sharp angular distribution of emission through the metal and down from the bottom of the structure. This coupling occurs over large distances to several hundred nm away from the metal and was found to be consistent with simulations of the reflectivity of the metal-dielectric waveguides. Remarkably, for some silica thicknesses, the emission is almost completely coupled through the structure with little free-space emission away from the metal-dielectric waveguide. The efficiency of fluorophore coupling is related to the quality of the resonant modes sustained by the metal-dielectric waveguide, resulting in coupling of most of the emission through the metal into the underlying glass substrates. Metal-dielectric waveguides also provide a method to resolve the emission from surface-bound fluorophores from the bulk-phase fluorophores. Metal-dielectric waveguides are simple to fabricate for large surface areas, the resonance wavelength can be adjusted by the dielectric thickness, and the silica surface is suitable for coupling to biomolecules. Metal-dielectric waveguides can have numerous applications in diagnostics and high-throughput proteomics or DNA analysis.

Dataset Information

Spectrally coded optical nanosectioning (SpecON) with biocompatible metal-dielectric-coated substrates.

Publications

Spectrally coded optical nanosectioning (SpecON) with biocompatible metal-dielectric-coated substrates.

Similar Datasets

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