title: Molecules and methods for super-resolution imaging.
abstract: By looking at a fluorescently labeled structure one molecule at a time, it is possible to side-step the optical diffraction limit and obtain "super-resolution" images of small nanostructures. In the Moerner Lab, we seek to develop both molecules and methods to extend super-resolution fluorescence imaging. Methodologies and protocols for designing and characterizing fluorophores with switchable fluorescence required for super-resolution imaging are reported. These fluorophores include azido-DCDHF molecules, covalently linked Cy3-Cy5 dimers, and also the first example of a photoswitchable fluorescent protein, enhanced yellow fluorescent protein (EYFP). The imaging of protein superstructures in living Caulobacter crescentus bacteria is used as an example of the power of super-resolution imaging by single-molecule photoswitching to extract information beyond the diffraction limit. Finally, a new method is described for obtaining three-dimensional super-resolution information using a double-helix point-spread function.
authors.affiliation: Department of Chemistry, Stanford University, Stanford, California, USA.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Thompson
authors.forename: Julie S
authors.initials: JS
authors.lastname: Biteen
authors.forename: Samuel J
authors.initials: SJ
authors.lastname: Lord
authors.forename: Nicholas R
authors.initials: NR
authors.lastname: Conley
authors.forename: W E
authors.initials: WE
authors.lastname: Moerner
mesh_terms: D000818:Animals; D001386:Azides / Q000737:chemistry; D001419:Bacteria / Q000648:ultrastructure; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D021621:Imaging, Three-Dimensional; D008856:Microscopy, Fluorescence / Q000379:methods*; D015394:Molecular Structure; D010777:Photochemistry; D017785:Photons
chemical_list: D001386:Azides; D005456:Fluorescent Dyes