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
Dynamic super-resolution structured illumination imaging in the living brain.
Raphaël Turcotte,Raphaël Turcotte,Yajie Liang,Masashi Tanimoto,Qinrong Zhang,Ziwei Li,Minoru Koyama,Eric Betzig,Eric Betzig,Eric Betzig,Na Ji,Na Ji,Na Ji +12 more
TL;DR: With adaptive optics to correcting sample-induced optical aberrations and optimized image acquisition and reconstruction to combat sample motion, this work adapted super-resolution structured illumination microscopy to in vivo imaging in the brains of zebrafish larvae and mice and observed the dynamics of dendrites and dendritic spines at nanoscale resolution.
Journal ArticleDOI
Video-rate multi-color structured illumination microscopy with simultaneous real-time reconstruction.
Andreas Markwirth,Mario Lachetta,Viola Mönkemöller,Viola Mönkemöller,Rainer Heintzmann,Rainer Heintzmann,Wolfgang Hübner,Thomas R Huser,Marcel Müller,Marcel Müller +9 more
TL;DR: The authors optimise both acquisition and reconstruction software to achieve multicolour SR-SIM at video frame-rates with reconstructed images displaying with only milliseconds delay during the experiment, a first in super-resolved structured illumination microscopy.
Journal ArticleDOI
High-throughput, high-resolution deep learning microscopy based on registration-free generative adversarial network.
TL;DR: This work combines a generative adversarial network (GAN) with light microscopy to achieve deep learning super-resolution under a large field of view (FOV) and proposes an image degrading model to generate low resolution images for training, making this approach free from the complex image registration during training data set preparation.
Journal ArticleDOI
From single molecules to life: microscopy at the nanoscale.
TL;DR: A theoretical overview of the techniques and underlying physics are presented, followed by a practical guide to all of the facets involved in designing a super-resolution experiment, including an approachable explanation of the photochemistry involved, labeling methods available, and sample preparation procedures.
Journal ArticleDOI
Stochastic optical reconstruction microscopy (STORM) in comparison with stimulated emission depletion (STED) and other imaging methods
Johnny Tam,David Merino +1 more
TL;DR: The choice between STORM and STED will depend not only on the specific application, but also on the users' ability to understand and optimize the various parameters ranging from sample preparation to image acquisition, which determine the quality of the final image.
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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