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Breaking the Diffraction Barrier: Super-Resolution Imaging of Cells

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

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Breaking the Diffraction Barrier: Super-Resolution Imaging of Cells

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  1. Breaking the Diffraction Barrier:Super-Resolution Imaging of Cells Edgar Ferrer-Lorenzo, Nicole Gagnon, Anna Torre 1

  2. 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

  3. STORM fluorescence microscopy can overcome the diffraction limit A StORMimage is constructed from the localization of individual fluorescent molecules that are switched on and off using light of different colors 3

  4. STORM’s method improves the resolution of fluorescence microscopy 4

  5. 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

  6. Creating an image with STORM Huang et al., Cell 143: 1047-1058 (2010). 6

  7. Applications of STORM • Cell biology • Microbiology • Neurobiology 7

  8. STORM can resolve DNA structure Fluorophores bound to DNA fragment RecAcoated circular plasmid DNA 8 Rust et al., Nature Methods 3: 793-796 (2006).

  9. High resolution images of microtubules Bates et al., Science 317, 1749-1753 (2007). 9

  10. STORM can resolve 3D structure Bates et al., Science 317, 1749-1753 (2007). 10

  11. High resolution imaging of the action cytoskeletal network in neurons Xu et al., Science 339: 452-456 (2013). 11

  12. 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

  13. Conclusions • STORM has found a clever way to get around the diffraction limit of resolution • Imaging resolution down to 20 nm • High-resolution live-cell imaging, thus enabling discovery of internal causes or origins of processes • All with visible light! • Imaging speed can be improved 13

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