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

  title={Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy},
  author={M. G. L. Gustafsson},
  journal={Journal of Microscopy},
Lateral resolution that exceeds the classical diffraction limit by a factor of two is achieved by using spatially structured illumination in a wide‐field fluorescence microscope. The sample is illuminated with a series of excitation light patterns, which cause normally inaccessible high‐resolution information to be encoded into the observed image. The recorded images are linearly processed to extract the new information and produce a reconstruction with twice the normal resolution. Unlike… 

Superresolution Multidimensional Imaging with Structured Illumination Microscopy

The resolution of an optical microscope is fundamentally limited by diffraction. In a conventional wide-field fluorescence microscope, the resolution limit is at best 200 nm. However, modern

Three-dimensional resolution doubling in wide-field fluorescence microscopy by structured illumination.

This work describes how spatially structured illumination microscopy can be applied in three dimensions to double the axial as well as the lateral resolution, with true optical sectioning, and has produced the first light microscopy images of the synaptonemal complex in which the lateral elements are clearly resolved.

Effects of inhomogeneous fields in superresolving structured-illumination microscopy.

It is shown how the use of high numerical aperture microscope objectives in structured-illumination microscopy can provide relatively high-frequency illumination patterns that can become evanescently decaying and thus becomes inhomogeneous within a microscopically extended sample medium.

Microscopy using randomized speckle illumination

It is well known for structured illumination microscopy (SIM) that the lateral resolution by a factor of two beyond the classical diffraction limit is achieved using spatially structured illumination

Super-resolved spatial light interference microscopy.

By adding a grating to the optical path, the structured illumination technique can be used to improve the resolution by a factor of 2.5 through two crucial modifications, namely, one to the pupil plane of the objective and the other to the demodulation procedure.

Lateral resolution enhancement of confocal microscopy based on structured detection method with spatial light modulator.

This paper introduces a new method to achieve structured detection through using spatial light modulator (SLM) to improve it, and it can be found that coherent transfer function expands and the resolution is 1.6 times as large as that of conventional confocal microscope.

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

  • M. Gustafsson
  • Physics
    Proceedings of the National Academy of Sciences of the United States of America
  • 2005
Experimental results show that a 2D point resolution of <50 nm is possible on sufficiently bright and photostable samples, and a recently proposed method in which the nonlinearity arises from saturation of the excited state is experimentally demonstrated.



Subdiffraction resolution in far-field fluorescence microscopy.

The resolution limit of scanning far-field fluorescence microscopy is overcame by disabling the fluorescence from the outer part of the focal spot by a spatially offset pulse.

Three-dimensional imaging of biological specimens with standing wave fluorescence microscopy

Axial resolution in fluorescence microscopy can be improved significantly by using standing wave illumination to selectively excite planes within the depth of field of the microscope. When the

Method of obtaining optical sectioning by using structured light in a conventional microscope.

A simple method of obtaining optical sectioning in a conventional wide-field microscope by projecting a single-spatial-frequency grid pattern onto the object and processing images that are substantially similar to those obtained with confocal microscopes is described.

The Role of the Pinhole in Confocal Imaging Systems

Confocal scanning microscopes are particularly attractive by virtue of their enhanced lateral resolution, purely coherent image formation and optical sectioning (Wilson and Sheppard, 1984). It is

Superresolution three-dimensional images of fluorescence in cells with minimal light exposure.

An algorithm has been developed that produces highly accurate images of fluorescent probe distribution inside cells with minimal light exposure and a conventional light microscope that provides resolution nearly four times greater than that currently available from any fluorescence microscope.

Extended resolution fluorescence microscopy.

  • M. Gustafsson
  • Biology, Chemistry
    Current opinion in structural biology
  • 1999

Fluorescence microscopy in three dimensions.

Confocal fluorescent microscopy with a finite-sized circular detector

For a confocal fluorescent microscope with a finite-sized circular detector, the three-dimensional optical transfer function (OTF) for a thick object has been developed without the use of Stockseth’s

Handbook of Biological Confocal Microscopy

Methods for Three-Dimensional Imaging and Tutorial on Practical Confocal Microscopy and Use of the Confocal Test Specimen.