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
Content-aware image restoration: pushing the limits of fluorescence microscopy.
Martin Weigert,Uwe Schmidt,Tobias Boothe,Andreas Müller,Alexandr Dibrov,Akanksha Jain,Benjamin Wilhelm,Deborah Schmidt,Coleman Broaddus,Siân Culley,Siân Culley,Mauricio Rocha-Martins,Fabián Segovia-Miranda,Caren Norden,Ricardo Henriques,Ricardo Henriques,Marino Zerial,Michele Solimena,Jochen C. Rink,Pavel Tomancak,Loic Royer,Florian Jug,Eugene W. Myers,Eugene W. Myers +23 more
TL;DR: This work shows how content-aware image restoration based on deep learning extends the range of biological phenomena observable by microscopy by bypassing the trade-offs between imaging speed, resolution, and maximal light exposure that limit fluorescence imaging to enable discovery.
Journal ArticleDOI
Super-Resolution Video Microscopy of Live Cells by Structured Illumination
Peter Kner,Bryant B. Chhun,Eric R. Griffis,Lukman Winoto,Mats G. L. Gustafsson,Mats G. L. Gustafsson +5 more
TL;DR: A high-speed structured-illumination microscope capable of 100-nm resolution at frame rates up to 11 Hz for several hundred time points is demonstrated by video imaging of tubulin and kinesin dynamics in living Drosophila melanogaster S2 cells in the total internal reflection mode.
Journal ArticleDOI
Plk4-Induced Centriole Biogenesis in Human Cells
Julia Kleylein-Sohn,Jens Westendorf,Mikael Le Clech,Robert Habedanck,York-Dieter Stierhof,Erich A. Nigg +5 more
TL;DR: It is shown that overexpression of Polo-like kinase 4 (Plk4) in human cells induces centrosome amplification through the simultaneous generation of multiple procentrioles adjoining each parental centriole, and that centrioles elongate through insertion of alpha-/beta-tubulin underneath a CP110 cap.
Journal ArticleDOI
Measuring image resolution in optical nanoscopy.
Robert P. J. Nieuwenhuizen,Keith A. Lidke,Mark Bates,Daniela Leyton Puig,David Grunwald,Sjoerd Stallinga,Bernd Rieger +6 more
TL;DR: This work introduces a measure based on Fourier ring correlation (FRC) that can be computed directly from an image and demonstrates its validity and benefits on two-dimensional (2D) and 3D localization microscopy images of tubulin and actin filaments.
Journal ArticleDOI
Roadmap on structured light
Halina Rubinsztein-Dunlop,Andrew Forbes,Michael V Berry,Mark R. Dennis,David L. Andrews,Masud Mansuripur,Cornelia Denz,Christina Alpmann,Peter Banzer,Thomas Bauer,Ebrahim Karimi,Lorenzo Marrucci,Miles J. Padgett,Monika Ritsch-Marte,Natalia M. Litchinitser,Nicholas P. Bigelow,Carmelo Rosales-Guzmán,Aniceto Belmonte,Juan P. Torres,Tyler W. Neely,Mark Baker,Reuven Gordon,Alexander B. Stilgoe,Jacquiline Romero,Andrew White,Robert Fickler,Alan E. Willner,Guodong Xie,Benjamin J. McMorran,Andrew M. Weiner +29 more
TL;DR: In this paper, the key fields within structured light from the perspective of experts in those areas, providing insight into the current state and the challenges their respective fields face, as well as the exciting prospects for the future that are yet to be realized.
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