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

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.

SUBMITTER: Rust MJ

PROVIDER: S-EPMC2700296 | biostudies-literature | 2006 Oct

REPOSITORIES: biostudies-literature

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

Rust Michael J MJ Bates Mark M Zhuang Xiaowei X

Nature methods 20060809 10

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 resolut ...[more]

PMID: 16896339

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| S-EPMC5835206 | biostudies-literature

Three-dimensional super-resolution imaging by stochastic optical reconstruction microscopy.

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STochastic Optical Reconstruction Microscopy (STORM) reveals the nanoscale organization of pathological aggregates in human brain.

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Volumetric super-resolution imaging by serial ultrasectioning and stochastic optical reconstruction microscopy in mouse neural tissue.

Project description:Here, we present a protocol for collecting large-volume, four-color, single-molecule localization imaging data from neural tissue. We have applied this technique to map the location and identities of chemical synapses across whole cells in mouse retinae. Our sample preparation approach improves 3D STORM image quality by reducing tissue scattering, photobleaching, and optical distortions associated with deep imaging. This approach can be extended for use on other tissue types enabling life scientists to perform volumetric super-resolution imaging in diverse biological models. For complete details on the use and execution of this protocol, please refer to Sigal et al. (2015).

| S-EPMC8637648 | biostudies-literature

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