Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy.
TLDR
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 with strikingly increased clarity compared to both conventional and confocal microscopes.Abstract:
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 confocal microscopy, the resolution improvement is achieved with no need to discard any of the emission light. The method produces images of strikingly increased clarity compared to both conventional and confocal microscopes.read more
Citations
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Journal ArticleDOI
A universal moire effect and application in X-ray phase-contrast imaging
TL;DR: The universal moiré effect helps overcome limitations in sensitivity and dose efficiency and obviates the need to make hard x-ray absorption gratings of sub-micron periods and enables a polychromatic far-field interferometer (PFI) without absorbing gratings.
Journal ArticleDOI
Fluorescence endomicroscopy with structured illumination.
TL;DR: An endomicroscope apparatus that utilizes structured illumination to produce high resolution optically sectioned fluorescence images over a field of view of about 240 microm is presented and a strategy to largely suppress structured illumination artifacts that arise when imaging in thick tissue that exhibits significant out-of-focus background is presented.
Journal ArticleDOI
Fluorescence perturbation techniques to study mobility and molecular dynamics of proteins in live cells: FRAP, photoactivation, photoconversion, and FLIP.
TL;DR: The materials required for performing FRAP experiments on a confocal laser-scanning microscopes and the software for data analysis are discussed, and general considerations on how to performFRAP experiments as well as the necessary controls are described.
Journal ArticleDOI
Fluorescence microscopy: established and emerging methods, experimental strategies, and applications in immunology.
TL;DR: This review is intended to address situations where the signal is weak, which is important for emerging techniques stressing super‐resolution or live cell dynamics, but is less important for conventional applications such as indirect immunofluorescence.
Journal ArticleDOI
Frontiers in structured illumination microscopy
TL;DR: Structured illumination microscopy (SIM) has been widely used in the life sciences as discussed by the authors and has been shown to be a powerful and versatile superresolution method for biochemical processes in laboratories around the world.
References
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BookDOI
Handbook of biological confocal microscopy
TL;DR: Methods for Three-Dimensional Imaging and Tutorial on Practical Confocal Microscopy and Use of the Confocal Test Specimen.
Journal ArticleDOI
Method of obtaining optical sectioning by using structured light in a conventional microscope
TL;DR: 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.
Journal ArticleDOI
Subdiffraction resolution in far-field fluorescence microscopy.
Thomas A. Klar,Stefan W. Hell +1 more
TL;DR: 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.
Book ChapterDOI
Fluorescence microscopy in three dimensions.
TL;DR: This chapter has discussed the nature of image formation in three dimensions and dealt with several means to remove contaminating out-of-focus information and developed a method for extremely rapidly and accurately producing an in-focus, high-resolution "synthetic projection" image from a thick specimen.
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