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
More filters
Journal ArticleDOI
Time-lapse two-color 3D imaging of live cells with doubled resolution using structured illumination
TL;DR: A previously undescribed SIM setup that is fast enough to record 3D two-color datasets of living whole cells and shows volume rates as high as 4 s in one color and 8.5 s in two colors over tens of time points is reported.
Journal ArticleDOI
Noninvasive Imaging beyond the Diffraction Limit of 3D Dynamics in Thickly Fluorescent Specimens
Liang Gao,Lin Shao,Christopher D. Higgins,John S. Poulton,Mark Peifer,Michael W. Davidson,Xufeng Wu,Bob Goldstein,Eric Betzig +8 more
TL;DR: It is reported that rapid three-dimensional dynamics can be studied beyond the diffraction limit in thick or densely fluorescent living specimens over many time points by combining ultrathin planar illumination produced by scanned Bessel beams with super-resolution structured illumination microscopy.
Journal ArticleDOI
Fluorescent proteins for live-cell imaging with super-resolution
TL;DR: There is still room for further improvement of these important markers for live cell imaging, and special FP variants with low switching fatigue have been introduced in recent years.
Journal ArticleDOI
Expansion microscopy: principles and uses in biological research.
TL;DR: This Perspective reviews available methods and provides practical guidance for users of Expansion microscopy, which allows super-resolution images of diverse samples to be acquired on conventional microscopes, thus democratizing super- resolution imaging.
Journal ArticleDOI
Tracking single molecules at work in living cells
TL;DR: SMT methods are reviewed, the recent results obtained by SMT are summarized, related superresolution microscopy data is summarized, and special concerns when SMT applications are shifted from the in vitro paradigms to living cells are described.
References
More filters
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.
Related Papers (5)
Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM).
Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy
Stefan W. Hell,Jan Wichmann +1 more