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