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Journal of the Optical Society of America A

Journal of the Optical Society of America A


  • Vol. 16, Iss. 4 — Apr. 1, 1999
  • pp: 909–915

Far-field fluorescence microscopy beyond the diffraction limit

A. M. van Oijen, J. Köhler, J. Schmidt, M. Müller, and G. J. Brakenhoff  »View Author Affiliations

JOSA A, Vol. 16, Issue 4, pp. 909-915 (1999)

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A technique has been developed to obtain three-dimensional structural information on a length scale well below the Rayleigh length with conventional far-field optics. By spectrally selecting a single molecule with high-resolution laser spectroscopy and using a CCD camera to register the spatial distribution of the emitted photons in three dimensions, one can determine the position of a molecule with unprecedented accuracy. One can resolve details in the specimen with sub-diffraction-limited resolution in three dimensions by applying this procedure to as many molecules as are present in the same diffraction-limited volume and obtaining their mutual positions. The feasibility of this technique is demonstrated for the system of pentacene in p-terphenyl at cryogenic temperatures for which molecules were localized with an accuracy of better than 40 nm in the lateral and 100 nm in the axial directions.

© 1999 Optical Society of America

OCIS Codes
(100.6640) Image processing : Superresolution
(180.1790) Microscopy : Confocal microscopy
(180.2520) Microscopy : Fluorescence microscopy

Original Manuscript: August 12, 1998
Revised Manuscript: November 16, 1998
Manuscript Accepted: November 20, 1998
Published: April 1, 1999

A. M. van Oijen, J. Köhler, J. Schmidt, M. Müller, and G. J. Brakenhoff, "Far-field fluorescence microscopy beyond the diffraction limit," J. Opt. Soc. Am. A 16, 909-915 (1999)

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