title: Localization-based super-resolution imaging of cellular structures.
abstract: Fluorescence microscopy allows direct visualization of fluorescently tagged proteins within cells. However, the spatial resolution of conventional fluorescence microscopes is limited by diffraction to ~250 nm, prompting the development of super-resolution microscopy which offers resolution approaching the scale of single proteins, i.e., ~20 nm. Here, we describe protocols for single molecule localization-based super-resolution imaging, using focal adhesion proteins as an example and employing either photoswitchable fluorophores or photoactivatable fluorescent proteins. These protocols should also be easily adaptable to imaging a broad array of macromolecular assemblies in cells whose components can be fluorescently tagged and assemble into high density structures.
authors.affiliation: Department of Bioengineering, Mechanobiology Institute, National University of Singapore, Singapore, Singapore.
authors.forename: Pakorn
authors.initials: P
authors.lastname: Kanchanawong
authors.forename: Clare M
authors.initials: CM
authors.lastname: Waterman
mesh_terms: D002477:Cells / Q000648:ultrastructure*; D022001:Focal Adhesions / Q000737:chemistry; D008856:Microscopy, Fluorescence / Q000379:methods*; D008967:Molecular Biology / Q000379:methods*; D046912:Multiprotein Complexes / Q000737:chemistry / Q000302:isolation & purification / Q000648:ultrastructure; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D046912:Multiprotein Complexes; D011506:Proteins