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
Lateral superresolution using a posteriori phase shift estimation for a moving object: experimental results
TL;DR: Experimental verification of earlier simulations for phase shift estimation a posteriori of fluorescence microscopy images for an object having unknown, random translational motion is shown to obtain an artifact-free reconstruction having the expected superresolution.
Journal ArticleDOI
Organic fluorescent probes for stochastic optical reconstruction microscopy (STORM): Recent highlights and future possibilities
Soham Samanta,Wanjun Gong,Wen Li,Amit Sharma,Inseob Shim,Wei Zhang,Pintu Das,Wenhui Pan,Liwei Liu,Zhigang Yang,Junle Qu,Jong Seung Kim,Jong Seung Kim +12 more
TL;DR: In this paper, the authors present a review of the use of organic fluorescent dyes for super-resolution imaging in eukaryotic biology and provide a basis on which advanced organic fluorescent probes can be designed and developed in the near future.
Journal ArticleDOI
Construction of an instant structured illumination microscope.
Alistair Curd,Alexa J. Cleasby,Katarzyna Anna Makowska,Andrew York,Hari Shroff,Michelle Peckham +5 more
TL;DR: In this paper, the instant structured illumination microscopy (iSIM) was proposed to capture high-resolution images at fast frame rates in live cells, and the assembly of an iSIM is reasonably complex, involving the combination and alignment of many optical components.
Journal ArticleDOI
Point by Point: An Introductory Guide to Sample Preparation for Single-Molecule, Super-Resolution Fluorescence Microscopy.
TL;DR: In this protocol, detailed instructions on preparation of robust reference samples for super‐resolution microscopy, including the cytoskeleton (microtubules), membrane‐bound organelles (peroxisomes), and scaffold proteins (clathrin), are provided.
Book ChapterDOI
Resolution in optical microscopy.
TL;DR: This chapter describes resolution in optical microscopy with a useful tool for comparing the performance of optical microscopes is the point-spread function (PSF), which can be defined in two complementary forms.
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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