Breaking the Diffraction Barrier:Super-Resolution Imaging of Cells

Edgar Ferrer-Lorenzo, Nicole Gagnon, Anna Torre

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Optical microscopes have limitations

• Diffraction limited• Resolution depends

on the wavelength of light and diameter of lens of optical microscope

• Specimen not alive

The solution: STORM

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STORM fluorescence microscopy can overcome the diffraction limit

A StORM image is constructed from the localization of individual fluorescent

molecules that are switched on and off using light of different colors

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STORM’s method improves the resolution of fluorescence microscopy

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STORM provides sub-diffraction-limit image resolution

Rust et al., Nature Methods 3: 793-796 (2006). Betzig et al., Science 313: 1642-1645 (2006). 5

Creating an image with STORM

6Huang et al., Cell 143: 1047-1058 (2010).

Applications of STORM

• Cell biology• Microbiology• Neurobiology

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STORM can resolve DNA structure

Fluorophores bound to DNA fragment

RecA coated circular plasmid DNA

Rust et al., Nature Methods 3: 793-796 (2006). 8

High resolution images of microtubules

Bates et al., Science 317, 1749-1753 (2007). 9

STORM can resolve 3D structure

Bates et al., Science 317, 1749-1753 (2007). 10

High resolution imaging of the action cytoskeletal network in neurons

Xu et al., Science 339: 452-456 (2013). 11

STORM has advanced biology

• Unambiguous identification of specific proteins

• Protein-protein interactions• Structure of small-type protein complexes• Live cell dynamics• Single molecule tracking• Cluster analysis and molecular counting

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Conclusions

1. STORM has found a clever way to get around the diffraction limit of resolution

2. Imaging resolution down to 20 nm

3. High-resolution live-cell imaging, thus enabling discovery of internal causes or origins of processes

4. All with visible light!

5. Imaging speed can be improved

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