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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Proceedings ArticleDOI
Closed loop adaptive optics for microscopy without a wavefront sensor.
TL;DR: This technique will be useful for aligning the deformable mirror in a widefield microscope with adaptive optics and could potentially be used to correct aberrations in samples where small fluorescent beads or other point sources are used as reference beacons.
Journal ArticleDOI
Eliminating deformations in fluorescence emission difference microscopy.
TL;DR: In deformation-free imaging, the resolution is higher than that of confocal imaging by 32%.
Journal ArticleDOI
Nanoscale imaging of clinical specimens using conventional and rapid-expansion pathology
Octavian Bucur,Feifei Fu,Mike Calderon,Geetha H. Mylvaganam,Geetha H. Mylvaganam,Ngoc L. Ly,Ngoc L. Ly,Jimmy Day,Simon Watkin,Bruce D. Walker,Bruce D. Walker,Bruce D. Walker,Edward S. Boyden,Yongxin Zhao,Yongxin Zhao +14 more
TL;DR: In this paper, the authors describe protocols for the simple and effective physical expansion of a variety of human tissues and clinical specimens, including paraffin-embedded, fresh frozen and chemically stained human tissues.
Journal ArticleDOI
Sub-diffraction Limit Localization of Proteins in Volumetric Space Using Bayesian Restoration of Fluorescence Images from Ultrathin Specimens
Gordon X. Wang,Stephen J. Smith +1 more
TL;DR: It is shown that AT with deconvolution increases the volumetric analytical fidelity of protein localization by significantly improving the modulation of high spatial frequencies up to and potentially beyond the spatial frequency cut-off of the objective.
Journal ArticleDOI
Feature-specific structured imaging
TL;DR: Compared to the optimal LMMSE postprocessing of a conventional image, feature-specific structured imaging provides a 38% rms error reduction and requires 400 times fewer measurements for a noise standard deviation of sigma = 2 x 10(-3).
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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