Nonlinear structured-illumination microscopy: Wide-field fluorescence imaging with theoretically unlimited resolution
Reads0
Chats0
TLDR
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.Abstract:
Contrary to the well known diffraction limit, the fluorescence microscope is in principle capable of unlimited resolution. The necessary elements are spatially structured illumination light and a nonlinear dependence of the fluorescence emission rate on the illumination intensity. As an example of this concept, this article experimentally demonstrates saturated structured-illumination microscopy, a recently proposed method in which the nonlinearity arisesfromsaturationoftheexcitedstate.Thismethodcanbeused in a simple, wide-field (nonscanning) microscope, uses only a single, inexpensive laser, and requires no unusual photophysical properties of the fluorophore. The practical resolving power is determined by the signal-to-noise ratio, which in turn is limited by photobleaching. Experimental results show that a 2D point resolution of <50 nm is possible on sufficiently bright and photostable samples.read more
Citations
More filters
Journal ArticleDOI
Subdiffraction-resolution fluorescence microscopy of myosin-actin motility.
TL;DR: A novel approach for video-like subdiffraction microscopy based on rapid and reversible photoswitching of commercially available organic carbocyanine fluorophores is introduced and in vitro studies on the motility of fluorophore-labeled actin filaments along myosin II are demonstrated.
Patent
Method and Configuration for the Optical Detection of an Illuminated Specimen
TL;DR: In this article, a method for the optical detection of an illuminated specimen is proposed, wherein the illuminating light impinges in a spatially structured manner in at least one plane on the specimen and several images of the specimen are acquired by a detector in different positions of the structure on the object.
Journal ArticleDOI
Modern Statistical Challenges in High-Resolution Fluorescence Microscopy
TL;DR: Super-resolution fluorescence microscopy has become an indispensable tool for studying the structure and dynamics of living organisms as mentioned in this paper, and this technique is inherently of a non-Gaussian statistical nature.
Journal ArticleDOI
Probing plasma membrane dynamics at the single-molecule level
TL;DR: Single-molecule imaging methods are reviewed, including total internal reflection fluorescence microscopy (TIRFM), fluorescence correlation spectroscopy (FCS), and super-resolution microscopy, and examine their contributions to recent progress in understanding protein dynamics and membrane organization in living plant cells.
Journal ArticleDOI
Super‐Resolution Microscopy: Going Live and Going Fast
TL;DR: This minireview focuses on the recent developments in live-cell super-resolution microscopy and its biological applications, enabling researchers to probe dynamic processes in living cells with unprecedented spatiotemporal resolution.
References
More filters
Journal ArticleDOI
Introduction to Fourier Optics
Joseph W. Goodman,Mary E. Cox +1 more
TL;DR: The second edition of this respected text considerably expands the original and reflects the tremendous advances made in the discipline since 1968 as discussed by the authors, with a special emphasis on applications to diffraction, imaging, optical data processing, and holography.
Book
Introduction to Fourier optics
TL;DR: The second edition of this respected text considerably expands the original and reflects the tremendous advances made in the discipline since 1968 as discussed by the authors, with a special emphasis on applications to diffraction, imaging, optical data processing, and holography.
Journal ArticleDOI
Two-Photon Laser Scanning Fluorescence Microscopy
TL;DR: The fluorescence emission increased quadratically with the excitation intensity so that fluorescence and photo-bleaching were confined to the vicinity of the focal plane as expected for cooperative two-photon excitation.
Journal ArticleDOI
Semiconductor Nanocrystals as Fluorescent Biological Labels
TL;DR: Semiconductor nanocrystals prepared for use as fluorescent probes in biological staining and diagnostics have a narrow, tunable, symmetric emission spectrum and are photochemically stable.
Book
The Principles of Nonlinear Optics
TL;DR: In this article, the authors present a general description of wave propagation in nonlinear media, including high-resolution nonlinear optical spectroscopy, and four-wave mixing and mixing.
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