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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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Journal ArticleDOI

ThunderSTORM: a comprehensive ImageJ plug-in for PALM and STORM data analysis and super-resolution imaging

TL;DR: ThunderSTORM is an open-source, interactive and modular plug-in for ImageJ designed for automated processing, analysis and visualization of data acquired by single-molecule localization microscope methods such as photo-activated localization microscopy and stochastic optical reconstruction microscopy.
Journal ArticleDOI

Revitalizing membrane rafts: new tools and insights

TL;DR: How the field has matured and an evolving model in which membranes are occupied by fluctuating nanoscale assemblies of sphingolipids, cholesterol and proteins that can be stabilized into platforms that are important in signalling, viral infection and membrane trafficking are presented.
Journal ArticleDOI

Subdiffraction Multicolor Imaging of the Nuclear Periphery with 3D Structured Illumination Microscopy

TL;DR: Three-dimensional structured illumination microscopy (3D-SIM) opens new and facile possibilities to analyze subcellular structures beyond the diffraction limit of the emitted light.
Journal ArticleDOI

Surface-Enhanced Raman Spectroscopy for Bioanalysis: Reliability and Challenges

TL;DR: An outlook of the key challenges in bioanalytical SERS, including reproducibility, sensitivity, and spatial and time resolution is given.
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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