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Breaking the Diffraction Barrier:Super-Resolution Imaging of Cells
Edgar Ferrer-Lorenzo, Nicole Gagnon, Anna Torre
1
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
2
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
3
STORM’s method improves the resolution of fluorescence microscopy
4
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
7
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
12
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
13