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
Image scanning microscopy: an overview.
Edward N. Ward,Robert Pal +1 more
TL;DR: A brief overview of the SIM and ISM processes and subsequent developments in the image reconstruction process is given, incorporating more recent achievements in light shaping (i.e. pattern scanning and super‐resolution beam shaping), to suggest potential future directions for this ever‐expanding field.
Journal ArticleDOI
Molecular strategies to read and write at the nanoscale with far-field optics
TL;DR: These promising far-field optical methods permit the convenient imaging of biological samples and fabrication of miniaturized objects with unprecedented resolution and can have long-term and profound implications in biomedical research and information technology.
Journal ArticleDOI
Resolution enhancement in nonlinear scanning microscopy through post-detection digital computation
TL;DR: In this paper, the authors investigated whether, as in wide field acquisition, significant resolution enhancement can be obtained by harnessing the nonlinear response of the sample when point-scanning structured illumination is employed.
Journal ArticleDOI
Sub-diffraction imaging on standard microscopes through photobleaching microscopy with non-linear processing.
Sebastian Munck,Katarzyna Miskiewicz,Ragna Sannerud,S. A. Menchón,Liya E. Jose,Rainer Heintzmann,Patrik Verstreken,Wim Annaert +7 more
TL;DR: This work presents a novel approach using photobleaching microscopy with non-linear processing (PiMP) for sub-diffraction imaging of cells and tissues using common fluorophores and can be implemented on standard wide-field or confocal systems.
Journal ArticleDOI
Dual-mode optical microscope based on single-pixel imaging
TL;DR: In this paper, the authors demonstrate an inverted microscope that can image specimens in both reflection and transmission modes simultaneously with a single light source. The system, a scanless device with no moving parts, works by sequential projection of a set of binary intensity patterns onto the sample that are codified onto a modified commercial DMD.
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