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Title:
Nonlinear structured-illumination microscopy: Wide-field fluorescence imaging with theoretically unlimited resolution
Authors:
Gustafsson, Mats G. L.
Affiliation:
Department of Physiology and Program in Bioengineering, University of California, San Francisco, CA 94143-2532
Publication:
Proceedings of the National Academy of Sciences, Volume 102, Issue 37, pp. 13081-13086.
Publication Date:
09/2005
Category:
BIOPHYSICS, BIOLOGICAL SCIENCES
Origin:
PNAS
Keywords:
super resolution, moiré, resolution extension, saturation
Abstract Copyright:
(c) 2005: The National Academy of Sciences
DOI:
10.1073/pnas.0406877102
Bibliographic Code:
2005PNAS..10213081G

Abstract

Contrary to the well known diffraction limit, the fluorescence microscope is in principle capable of unlimited resolution. The necessary elements are spatially structured illumination light and a nonlinear dependence of the fluorescence emission rate on the illumination intensity. As an example of this concept, this article experimentally demonstrates saturated structured-illumination microscopy, a recently proposed method in which the nonlinearity arises from saturation of the excited state. This method can be used in a simple, wide-field (nonscanning) microscope, uses only a single, inexpensive laser, and requires no unusual photophysical properties of the fluorophore. The practical resolving power is determined by the signal-to-noise ratio, which in turn is limited by photobleaching. Experimental results show that a 2D point resolution of <50 nm is possible on sufficiently bright and photostable samples.

Author contributions: M.G.L.G. designed research, performed research, analyzed data, and wrote the paper.This paper was submitted directly (Track II) to the PNAS office.Abbreviations: SSIM, saturated structured-illumination microscopy; FWHM, full width at half maximum.

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