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We report the breaking of the diffraction resolution barrier in far-field fluorescence microscopy by transiently shelving the fluorophore in a metastable dark state. Using a relatively modest light intensity of several kW/cm(2) in a focal distribution featuring a local zero, we confine the fluorescence emission to a spot whose diameter is a fraction of the wavelength of light. Nanoscale far-field optical resolution down to 50 nm is demonstrated by imaging microtubules in a mammalian cell and proteins on the plasma membrane of a neuron. The presence of dark states in virtually any fluorescent molecule opens up a new venue for far-field microscopy with resolution that is no longer limited by diffraction.

Original publication

DOI

10.1103/PhysRevLett.98.218103

Type

Journal article

Journal

Phys Rev Lett

Publication Date

25/05/2007

Volume

98

Keywords

Cell Line, Fluorescent Dyes, Humans, Microscopy, Fluorescence, Optics and Photonics, Synaptosomal-Associated Protein 25