Self-interference fluorescent emission microscopy 5-nm vertical resolution

Swan, Anna K.; Ünlü, M. Selim; Tong, Yunjie; Goldberg, Bennett B.; Moiseev, L.; Cantor, Charles R.

doi:10.1109/cleo.2001.947913

Cited by 2 publications

(1 citation statement)

References 2 publications

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“…The broader the emission spectrum, the more information is collected and the more precise the height determination. The distance above the mirror can be determined solely from the oscillations within the spectrum [20], [21]. Tanigushi et al [22] observed this type of oscillations for a broad distribution of vertical positions of fluorophores and found a qualitative agreement with data using a classical model of self-interference of the spontaneous emission.…”

Section: Spectral Self-interference Fluorescence Microscopymentioning

confidence: 91%

Toward nanometer-scale resolution in fluorescence microscopy using spectral self-interference

Swan

Moiseev

Cantor

et al. 2003

IEEE J. Select. Topics Quantum Electron.

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We introduce a new fluorescence microscopy technique that maps the axial position of a fluorophore with subnanometer precision. The interference of the emission of fluorophores in proximity to a reflecting surface results in fringes in the fluorescence spectrum that provide a unique signature of the axial position of the fluorophore. The nanometer sensitivity is demonstrated by measuring the height of a fluorescein monolayer covering a 12-nm step etched in silicon dioxide. In addition, the separation between fluorophores attached to the top or the bottom layer in a lipid bilayer film is determined. We further discuss extension of this microscopy technique to provide resolution of multiple layers spaced as closely as 10 nm for sparse systems.Index Terms-Fluorescence microscopy, interference, spectroscopy, ultra high-optical resolution.

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Section: Spectral Self-interference Fluorescence Microscopymentioning

confidence: 91%

Toward nanometer-scale resolution in fluorescence microscopy using spectral self-interference

Swan

Moiseev

Cantor

et al. 2003

IEEE J. Select. Topics Quantum Electron.

Self Cite

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High-resolution spectral self-interference fluorescence microscopy

Swan

Moiseev

Tong³

et al. 2002

SPIE Proceedings

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We present a new method of fluorescence imaging, which yields nm-scale axial height determination and ~15 nm axial resolution. The method uses the unique spectral signature of the fluorescent emission intensity well above a reflecting surface to determine vertical position unambiguously. We have demonstrated axial height determination with nm sensitivity by resolving the height difference of fluorescein directly on the surface or ontop of streptavidin. While different positions of fluorophores of different color are determined independently with nm precision, resolving the position of two fluorophores of the same color is a more convoluted problem due to the finite spectral emission widow of the fluorophores. Hence, for physically close (<λ/2) fluorophores, it is necessary to collect multiple spectra by independently scanning an excitation standing wave in order to deconvolute the contribution to the spectral pattern from different heights. Moving the excitation standing wave successively enhances or suppresses excitation from different parts of the height distribution, changing the spectral content. This way two fluorophores of the same color can be resolved to better than 20 nm. Design aspects of the dielectric stack for independent excitation wave scanning and limits of deconvolution for an arbitrary height distribution will be discussed.

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Self-interference fluorescent emission microscopy 5-nm vertical resolution

Cited by 2 publications

References 2 publications

Toward nanometer-scale resolution in fluorescence microscopy using spectral self-interference

Toward nanometer-scale resolution in fluorescence microscopy using spectral self-interference

High-resolution spectral self-interference fluorescence microscopy

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