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Far-field fluorescence microscopy beyond the diffraction limit

Journal Article


Abstract


  • 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 highresolution 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.

Publication Date


  • 1999

Citation


  • van Oijen, A. M., Köhler, J., Schmidt, J., Müller, M., & Brakenhoff, G. J. (1999). Far-field fluorescence microscopy beyond the diffraction limit. Journal of the Optical Society of America A: Optics and Image Science, and Vision, 16(4), 909-915. doi:10.1364/JOSAA.16.000909

Scopus Eid


  • 2-s2.0-35848970592

Start Page


  • 909

End Page


  • 915

Volume


  • 16

Issue


  • 4

Abstract


  • 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 highresolution 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.

Publication Date


  • 1999

Citation


  • van Oijen, A. M., Köhler, J., Schmidt, J., Müller, M., & Brakenhoff, G. J. (1999). Far-field fluorescence microscopy beyond the diffraction limit. Journal of the Optical Society of America A: Optics and Image Science, and Vision, 16(4), 909-915. doi:10.1364/JOSAA.16.000909

Scopus Eid


  • 2-s2.0-35848970592

Start Page


  • 909

End Page


  • 915

Volume


  • 16

Issue


  • 4