Breaking the diffraction barrier in fluorescence microscopy by optical shelving - PubMed (original) (raw)

. 2007 May 25;98(21):218103.

doi: 10.1103/PhysRevLett.98.218103. Epub 2007 May 24.

Affiliations

• PMID: 17677813
• DOI: 10.1103/PhysRevLett.98.218103

Breaking the diffraction barrier in fluorescence microscopy by optical shelving

Stefan Bretschneider et al. Phys Rev Lett. 2007.

Abstract

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.

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