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
Imaging intracellular fluorescent proteins at nanometer resolution.
Eric Betzig,George H. Patterson,Rachid Sougrat,O. Wolf Lindwasser,Scott G. Olenych,Juan S. Bonifacino,Michael W. Davidson,Jennifer Lippincott-Schwartz,Harald F. Hess +8 more
TL;DR: This work introduced a method for optically imaging intracellular proteins at nanometer spatial resolution and used this method to image specific target proteins in thin sections of lysosomes and mitochondria and in fixed whole cells to image retroviral protein Gag at the plasma membrane.
Journal ArticleDOI
Microglia sculpt postnatal neural circuits in an activity and complement-dependent manner.
Dorothy P. Schafer,Emily K. Lehrman,Amanda G. Kautzman,Ryuta Koyama,Alan R. Mardinly,Ryo Yamasaki,Richard M. Ransohoff,Michael E. Greenberg,Ben A. Barres,Beth Stevens +9 more
TL;DR: It is shown that microglia engulf presynaptic inputs during peak retinogeniculate pruning and that engulfment is dependent upon neural activity and themicroglia-specific phagocytic signaling pathway, complement receptor 3(CR3)/C3.
Journal ArticleDOI
Far-Field Optical Nanoscopy
TL;DR: Initial applications indicate that emergent far-field optical nanoscopy will have a strong impact in the life sciences and in other areas benefiting from nanoscale visualization.
Journal ArticleDOI
Nonlinear structured-illumination microscopy: Wide-field fluorescence imaging with theoretically unlimited resolution
TL;DR: Experimental results show that a 2D point resolution of <50 nm is possible on sufficiently bright and photostable samples, and a recently proposed method in which the nonlinearity arises from saturation of the excited state is experimentally demonstrated.
Journal ArticleDOI
Super-Resolution Fluorescence Microscopy
TL;DR: It is anticipated that super-resolution fluorescence microscopy will become a widely used tool for cell and tissue imaging to provide previously unobserved details of biological structures and processes.
References
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Proceedings ArticleDOI
Sevenfold improvement of axial resolution in 3D wide-field microscopy using two objective lenses
TL;DR: In this paper, the authors proposed two wide field techniques with substantially improved axial resolution that actually exceeds the lateral resolution, which is related to the angle over which the objective lens collects light.
Book ChapterDOI
The Role of the Pinhole in Confocal Imaging Systems
TL;DR: Confocal scanning microscopes are particularly attractive by virtue of their enhanced lateral resolution, purely coherent image formation and optical sectioning and it is probably the latter property which is most useful as it gives rise to the ability to image a thick specimen in three-dimensions.
Journal ArticleDOI
Real time 3D fluorescence microscopy by two beam interference illumination
TL;DR: In this article, a method of obtaining optically sectioned fluorescence images in a widefield conventional microscope by interfering two beams on an object so as to illuminate it with a single spatial frequency fringe pattern is described.
Journal ArticleDOI
Confocal fluorescent microscopy with a finite-sized circular detector
Min Gu,Colin J. R. Sheppard +1 more
TL;DR: For a confocal fluorescent microscope with a finite-sized circular detector, the three-dimensional optical transfer function for a thick object has been developed without the use of Stockseth's approximation.
Proceedings ArticleDOI
Three-dimensional imaging of biological specimens with standing wave fluorescence microscopy
TL;DR: In this article, standing wave illumination was used to selectively excite planes within the depth of field of the microscope to estimate the 3D structure of a 3D object from three images within the same focal plane.
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