Breaking Abbe’s diffraction resolution limit in fluorescence microscopy with stimulated emission depletion beams of various shapes

Thomas A. Klar, Egbert Engel, and Stefan W. Hell
Phys. Rev. E 64, 066613 – Published 26 November 2001
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Abstract

We report on the generation of various hole-centered beams in the focal region of a lens and investigate their effectiveness to break the diffraction barrier in fluorescence microscopy by stimulated emission. Patterning of the phase of the stimulating beam across the entrance pupil of the objective lens produces point-spread-functions with twofold, fourfold, and circular symmetry, which narrow down the focal spot to 65–100 nm. Comparison with high-resolution confocal images exhibits a resolution much beyond the diffraction barrier. Particles that are only 65-nm apart are resolved with focused light.

  • Received 7 May 2001

DOI:https://doi.org/10.1103/PhysRevE.64.066613

©2001 American Physical Society

Authors & Affiliations

Thomas A. Klar, Egbert Engel, and Stefan W. Hell*

  • High Resolution Optical Microscopy Group, Max-Planck-Institute for Biophysical Chemistry, 37070 Göttingen, Germany

  • *Email address: hell@4pi.de

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Issue

Vol. 64, Iss. 6 — December 2001

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