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.
Keywords
PhotobleachingMicroscopyMicroscopeFluorophoreOpticsResolution (logic)FluorescenceMaterials scienceLaserNonlinear systemPhysicsComputer science
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Publication Info
- Year
- 2005
- Type
- article
- Volume
- 102
- Issue
- 37
- Pages
- 13081-13086
- Citations
- 2187
- Access
- Closed
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Cite This
Mats G. Gustafsson
(2005).
Nonlinear structured-illumination microscopy: Wide-field fluorescence imaging with theoretically unlimited resolution.
Proceedings of the National Academy of Sciences
, 102
(37)
, 13081-13086.
https://doi.org/10.1073/pnas.0406877102
Identifiers
- DOI
- 10.1073/pnas.0406877102