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
Potential Role for a Specialized β3 Integrin-Based Structure On Osteocyte Processes in Bone Mechanosensation†
Pamela Cabahug-Zuckerman,Randy F. Stout,Randy F. Stout,Robert J. Majeska,Mia M. Thi,David C. Spray,Sheldon Weinbaum,Mitchell B. Schaffler +7 more
TL;DR: The hypothesis that in authentic osteocytes in situ, key membrane proteins implicated in osteocyte mechanotransduction are preferentially localized at or near to β3 integrin‐foci is tested and identified a possible structural basis for the unique mechanosensation and transduction capabilities of the osteocyte process.
Journal ArticleDOI
3D sub-diffraction imaging in a conventional confocal configuration by exploiting super-linear emitters.
Denitza Denkova,Martin Ploschner,Martin Ploschner,Minakshi Das,Lindsay M. Parker,Xianlin Zheng,Yiqing Lu,Antony Orth,Nicolle H. Packer,Nicolle H. Packer,James A. Piper +10 more
TL;DR: A conceptually different super-resolution approach which circumvents limitations and enables 3D sub-diffraction imaging on conventional confocal microscopes and relies on markers with super-linear dependence of the emission on the excitation power.
Journal ArticleDOI
A Fluorescent Indicator for Imaging Lysosomal Zinc(II) with Förster Resonance Energy Transfer (FRET)‐Enhanced Photostability and a Narrow Band of Emission
TL;DR: It was shown using two-color structured illumination microscopy (SIM), which is capable of extending the lateral resolution over the Abbe diffraction limit by a factor of two, that the morpholino-functionalized compound 4 localizes in the interior of lysosomes, rather than anchoring on the lysOSomal membranes, of live HeLa cells.
Journal ArticleDOI
Imaging Living Synapses at the Nanoscale by STED Microscopy
U. V. Nagerl,Tobias Bonhoeffer +1 more
TL;DR: One of the principle goals in this research was to establish a novel type of synapticplasticity in the central nervous system by exploiting the role of acetylcholine in the regulation of cell reprograming.
Journal ArticleDOI
Fluorescence microscopy below the diffraction limit.
TL;DR: This chapter provides an overview of these new "super-resolution" imaging methods available that surpass this barrier and improve resolution up to 10 times over that of conventional microscopy.
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