Fast three-dimensional super-resolution imaging of live cells

By S. A. Jones, S. Shim, Jiang He, X. Zhuang. Nature Methods, 2011

718 citations recorded by the source corpus.

We report super-resolution fluorescence imaging of live cells with high spatiotemporal resolution using stochastic optical reconstruction microscopy (STORM). By labeling proteins either directly or via SNAP tags with photoswitchable dyes, we obtained two-dimensional (2D) and 3D super-resolution images of living cells, using clathrin-coated pits and the transferrin cargo as model systems. Bright, fast-switching probes enabled us to achieve 2D imaging at spatial resolutions of ∼25 nm and temporal resolutions as fast as 0.5 s. We also demonstrated live-cell 3D super-resolution imaging. We obtained 3D spatial resolution of ∼30 nm in the lateral direction and ∼50 nm in the axial direction at time resolutions as fast as 1–2 s with several independent snapshots. Using photoswitchable dyes with distinct emission wavelengths, we also demonstrated two-color 3D super-resolution imaging in live cells. These imaging capabilities open a new window for characterizing cellular structures in living cells at the ultrastructural level.

Subfields: Atomic and Molecular Physics, and Optics, Biophysics

View Paper: https://www.semanticscholar.org/paper/28ccebb047d6000d0280d8bc147cc15fd214858e

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In Biophysics

Citations in the corpus

Biological Imaging Software Tools (2012, methodology): “Examples of developments enabled by software written in research laboratories include structured-illumination microscopy 2 , super-resolution microscopy 3–5 and Bessel-beam microscopy 6 .”
Deep learning massively accelerates super-resolution localization microscopy (2018, background): “…dyes with rapid switching kinetics, high-power lasers, and fast cameras allows minimization of exposure time without decreasing the signal-to-noise ratio 5,6 , but reaching submil-lisecond exposure remains challenging, and intense irradiation exacerbates phototoxicity in live-cell imaging 7,8 .”
Evaluation of fluorophores for optimal performance in localization-based super-resolution imaging (2011, neutral): “When a photoswitchable reporter alone is used for STORM imaging, the activation rate is typically controlled by an ultraviolet or violet light 10,20,39 .”