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Open AccessJournal ArticleDOI

Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM).

TLDR
A high-resolution fluorescence microscopy method based on high-accuracy localization of photoswitchable fluorophores that can, in principle, reach molecular-scale resolution is developed.
Abstract
We have developed a high-resolution fluorescence microscopy method based on high-accuracy localization of photoswitchable fluorophores. In each imaging cycle, only a fraction of the fluorophores were turned on, allowing their positions to be determined with nanometer accuracy. The fluorophore positions obtained from a series of imaging cycles were used to reconstruct the overall image. We demonstrated an imaging resolution of 20 nm. This technique can, in principle, reach molecular-scale resolution.

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Spatially resolved, highly multiplexed RNA profiling in single cells

TL;DR: This report reports multiplexed error-robust FISH (MERFISH), a single-molecule imaging method that allows thousands of RNA species to be imaged in single cells by using combinatorial FISH labeling with encoding schemes capable of detecting and/or correcting errors.
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.
Journal ArticleDOI

Multicolor Super-Resolution Imaging with Photo-Switchable Fluorescent Probes

TL;DR: A family of photo-switchable fluorescent probes is introduced and multicolor stochastic optical reconstruction microscopy (STORM) is demonstrated, to facilitate direct visualization of molecular interactions at the nanometer scale.
Journal ArticleDOI

Spiropyran-based dynamic materials

TL;DR: This review discusses the synthesis, switching conditions, and use of dynamic materials in which spiropyran has been attached to the surfaces of polymers, biomacromolecules, inorganic nanoparticles, as well as solid surfaces.
References
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Journal ArticleDOI

Nonlinear magic: multiphoton microscopy in the biosciences

TL;DR: Multiphoton microscopy has found a niche in the world of biological imaging as the best noninvasive means of fluorescence microscopy in tissue explants and living animals and its use is now increasing exponentially.
Journal ArticleDOI

Precise nanometer localization analysis for individual fluorescent probes

TL;DR: A localization algorithm motivated from least-squares fitting theory is constructed and tested both on image stacks of 30-nm fluorescent beads and on computer-generated images (Monte Carlo simulations), and results show good agreement with the derived precision equation.
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.
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Myosin V Walks Hand-Over-Hand: Single Fluorophore Imaging with 1.5-nm Localization

TL;DR: The results strongly support a hand-over-hand model of motility, not an inchworm model, which moves processively on actin.
Journal ArticleDOI

Toward fluorescence nanoscopy

TL;DR: A family of concepts has emerged that overcomes the diffraction barrier altogether and, relying on saturated optical transitions, these concepts are limited only by the attainable saturation level.
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