title: Photoswitchable fluorophores for single-molecule localization microscopy.
abstract: Over the past decade, fluorescence microscopy has been revolutionized by the development of novel techniques that allow near-molecular resolution. Many such methods-collectively referred to as "single-molecule localization microscopy" (SMLM)-are based upon the repeated imaging of sparse stochastic subsets of the fluorophores in a sample. Active fluorophores are localized by finding the centers of their point spread functions, and a super-resolution image is constructed.Key to this strategy is the use of fluorophores that can be switched "on" and "off" in a controllable manner. Here we review the strengths and weaknesses of the wide variety of SMLM-compatible photoswitchable fluorophores and labeling strategies currently available. We also discuss their suitability for live-cell and multicolor imaging, as well as molecular counting.
authors.affiliation: Department of Biotechnology and Biophysics, Julius-Maximilians University Würzburg, Würzburg, Germany.
authors.forename: Kieran
authors.initials: K
authors.lastname: Finan
authors.forename: Benjamin
authors.initials: B
authors.lastname: Flottmann
authors.forename: Mike
authors.initials: M
authors.lastname: Heilemann
mesh_terms: D003116:Color; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods*; D057054:Molecular Imaging; D055620:Optical Phenomena; D013194:Staining and Labeling
chemical_list: D005456:Fluorescent Dyes; D008164:Luminescent Proteins