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Journal of the Optical Society of America A

Journal of the Optical Society of America A

| OPTICS, IMAGE SCIENCE, AND VISION

  • Vol. 19, Iss. 8 — Aug. 1, 2002
  • pp: 1599–1609

Saturated patterned excitation microscopy—a concept for optical resolution improvement

Rainer Heintzmann, Thomas M. Jovin, and Christoph Cremer  »View Author Affiliations


JOSA A, Vol. 19, Issue 8, pp. 1599-1609 (2002)
http://dx.doi.org/10.1364/JOSAA.19.001599


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Abstract

The resolution of optical microscopy is limited by the numerical aperture and the wavelength of light. Many strategies for improving resolution such as 4Pi and I5M have focused on an increase of the numerical aperture. Other approaches have based resolution improvement in fluorescence microscopy on the establishment of a nonlinear relationship between local excitation light intensity in the sample and in the emitted light. However, despite their innovative character, current techniques such as stimulated emission depletion (STED) and ground-state depletion (GSD) microscopy require complex optical configurations and instrumentation to narrow the point-spread function. We develop the theory of nonlinear patterned excitation microscopy for achieving a substantial improvement in resolution by deliberate saturation of the fluorophore excited state. The postacquisition manipulation of the acquired data is computationally more complex than in STED or GSD, but the experimental requirements are simple. Simulations comparing saturated patterned excitation microscopy with linear patterned excitation microscopy (also referred to in the literature as structured illumination or harmonic excitation light microscopy) and ordinary widefield microscopy are presented and discussed. The effects of photon noise are included in the simulations.

© 2002 Optical Society of America

OCIS Codes
(110.0180) Imaging systems : Microscopy
(110.2990) Imaging systems : Image formation theory
(110.4850) Imaging systems : Optical transfer functions
(170.0180) Medical optics and biotechnology : Microscopy
(180.2520) Microscopy : Fluorescence microscopy
(190.1900) Nonlinear optics : Diagnostic applications of nonlinear optics

History
Original Manuscript: July 31, 2001
Revised Manuscript: January 14, 2002
Manuscript Accepted: March 7, 2002
Published: August 1, 2002

Citation
Rainer Heintzmann, Thomas M. Jovin, and Christoph Cremer, "Saturated patterned excitation microscopy—a concept for optical resolution improvement," J. Opt. Soc. Am. A 19, 1599-1609 (2002)
http://www.opticsinfobase.org/josaa/abstract.cfm?URI=josaa-19-8-1599

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