title: Imaging cells at the nanoscale.
abstract: Recently developed super-resolution techniques in optical microscopy have pushed the length scale at which cellular structure can be observed down to tens of nanometres. A wide array of methods have been described that fall under the umbrella term of super-resolution microscopy and each of these methods has different requirements for acquisition speed, experimental complexity, fluorophore requirements and post-processing of data. For example, experimental complexity can be decreased by using a standard widefield microscope for acquisition, but this requires substantial processing of the data to extract the super-resolution information. These powerful techniques are bringing new insights into the nanoscale structure of sub-cellular assemblies such as podosomes, which are an ideal system to observe with super-resolution microscopy as the structures are relatively thin and they form and dissociate over a period of several minutes. Here we discuss the major classes of super-resolution microscopy techniques, and demonstrate their relative performance by imaging podosomes.
authors.affiliation: Randall Division of Cell & Molecular Biophysics, King's College London, London, UK. susan.cox@kcl.ac.uk
authors.forename: Susan
authors.initials: S
authors.lastname: Cox
authors.forename: Gareth E
authors.initials: GE
authors.lastname: Jones
mesh_terms: D000818:Animals; D002477:Cells / Q000166:cytology*; D006801:Humans; D008853:Microscopy; D057054:Molecular Imaging / Q000379:methods*; D036103:Nanotechnology / Q000379:methods*
keywords: 3B; Bayesian analysis of blinking and bleaching; Localisation microscopy; PALM; PSF; Podosome; SIM; SOFI; SSIM; STED; STORM; Stimulated emission depletion microscopy; Structured illumination microscopy; Super-resolution microscopy; TIRF; dSTORM; direct stochastic optical reconstruction microscopy; photoactivatable localisation microscopy; point spread function; saturated structured illumination microscopy; stimulated emission depletion microscopy; stochastic optical reconstruction microscopy; structured illumination microscopy; super-resolution optical fluctuation imaging; total internal reflectance microscopy