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      Nonlinear structured-illumination microscopy: Wide-field fluorescence imaging with theoretically unlimited resolution

      Proceedings of the National Academy of Sciences

      Proceedings of the National Academy of Sciences

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          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.

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          Most cited references 28

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          Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy.

          We propose a new type of scanning fluorescence microscope capable of resolving 35 nm in the far field. We overcome the diffraction resolution limit by employing stimulated emission to inhibit the fluorescence process in the outer regions of the excitation point-spread function. In contrast to near-field scanning optical microscopy, this method can produce three-dimensional images of translucent specimens.
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            Two-photon laser scanning fluorescence microscopy

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              Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy. SHORT COMMUNICATION

               M. Gustafsson (2000)
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                Author and article information

                Journal
                Proceedings of the National Academy of Sciences
                Proceedings of the National Academy of Sciences
                Proceedings of the National Academy of Sciences
                0027-8424
                1091-6490
                September 13 2005
                September 13 2005
                September 02 2005
                September 13 2005
                : 102
                : 37
                : 13081-13086
                10.1073/pnas.0406877102
                1201569
                16141335
                © 2005
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