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
Superresolution imaging of biological nanostructures by spectral precision distance microscopy.
Christoph Cremer,Rainer Kaufmann,Manuel Gunkel,Sebastian Pres,Yanina Weiland,Patrick Müller,Thomas Ruckelshausen,Paul Lemmer,Fania Geiger,Sven Degenhard,Christina Wege,Niels A. W. Lemmermann,Rafaela Holtappels,Hilmar Strickfaden,Michael Hausmann +14 more
TL;DR: The principles of spectrally assigned localization microscopy (SALM) of biological nanostructures are described, focusing on a special SALM approach, spectral precision distance/position determination microscopy(SPDM) with physically modified fluorochromes (SPDMPhymod), based on high‐precision localization of fluorescent molecules.
Journal ArticleDOI
STED microscopy of living cells--new frontiers in membrane and neurobiology.
TL;DR: The implementation of STED microscopy is presented for novel insights into live cell mechanisms, with a focus on neurobiology and plasma membrane dynamics.
Journal ArticleDOI
SIMToolbox: a MATLAB toolbox for structured illumination fluorescence microscopy
TL;DR: UNLABELLED SIMToolbox is an open-source, modular set of functions for MATLAB equipped with a user-friendly graphical interface and designed for processing two-dimensional and three-dimensional data acquired by structured illumination microscopy.
Journal ArticleDOI
Resolution enhancement of confocal microscopy by subtraction method with vector beams.
TL;DR: The subtraction imaging using vector beams demonstrated high spatial resolution with avoiding the negative side lobe and further resolution enhancement beyond 100 nm was predicted by using a flat-top beam obtained by the combination of beams with radial and azimuthal polarizations.
Journal ArticleDOI
Chemistry of Photosensitive Fluorophores for Single-Molecule Localization Microscopy
Fadi M. Jradi,Luke D. Lavis +1 more
TL;DR: The history of single-molecule localization microscopy is detailed and the collection of probes with demonstrated utility in SMLM is collated to serve as a primer for probe choice in localization microscope as well as an inspiration for the development of new fluorophores that enable imaging of biological samples with exquisite detail.
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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