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
Fast Axial-Scanning Widefield Microscopy With Constant Magnification and Resolution
Manuel Martínez-Corral,Po-Yuan Hsieh,Ana Doblas,Emilio Sánchez-Ortiga,Genaro Saavedra,Yi-Pai Huang +5 more
TL;DR: In this paper, the use of electrically-addressable lens devices for performing fast non-mechanical axial scanning when imaging three-dimensional samples was proposed, which is based on the insertion of the tunable lens at the aperture stop of the microscope objective.
Journal ArticleDOI
Signal, noise and resolution in linear and nonlinear structured-illumination microscopy
E.A. Ingerman,R.A. London,Rainer Heintzmann,Rainer Heintzmann,Mats G. L. Gustafsson,Mats G. L. Gustafsson +5 more
TL;DR: 1D patterns are advantageous in the linear case, and that in the nonlinear case 2D patterns provide a slight signal‐to‐noise advantage under idealised conditions, but perform worse than 1D patterns in the presence of nonswitchable fluorescent background.
Journal ArticleDOI
Quantitative optical microscope with enhanced resolution using a pixelated liquid crystal spatial light modulator
TL;DR: This paper presents a brief account of a novel optical microscope, which combines the advantages of two well‐known techniques, namely phase contrast and phase stepping, to provide high contrast imaging and precision measurements.
Journal ArticleDOI
Seeing beyond the limit: A guide to choosing the right super-resolution microscopy technique.
Jessica Valli,Adrian Garcia-Burgos,Liam M. Rooney,Beatriz Vale de Melo e Oliveira,Rory R. Duncan,Colin Rickman +5 more
TL;DR: In this article, the authors provide an overview and clarify the concepts underlying the most commonly available super-resolution techniques as well as guide researchers through all aspects that should be considered before opting for a given technique.
Journal ArticleDOI
Superresolution Localization Methods
TL;DR: This review examines several different families of fluorophore localization algorithms, comparing their complexity, performance, and range of applicability (e.g., whether they require particular types of experimental information, are optimized for specific situations, or are more general).
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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