title: Does super-resolution fluorescence microscopy obsolete previous microscopic approaches to protein co-localization?
abstract: Conventional microscopy techniques, namely, the confocal microscope or deconvolution processes, are resolution limited to approximately 200-250 nm by the diffraction properties of light as developed by Ernst Abbe in 1873. This diffraction limit is appreciably above the size of most multi-protein complexes, which are typically 20-50 nm in diameter. In the mid-2000s, biophysicists moved beyond the diffraction barrier by structuring the illumination pattern and then applying mathematical principles and algorithms to allow a resolution of approximately 100 nm, sufficient to address protein subcellular co-localization questions. This "breaking" of the diffraction barrier, affording resolution beyond 200 nm, is termed super-resolution microscopy. More recent approaches include single-molecule localization (such as photoactivated localization microscopy (PALM)/stochastic optical reconstruction microscopy (STORM)) and point spread function engineering (such as stimulated emission depletion (STED) microscopy). In this review, we explain basic principles behind currently commercialized super-resolution setups and address advantages and considerations in applying these techniques to protein co-localization in biological systems.
authors.affiliation: Department of Physiology and Biophysics, University of Arkansas for Medical Sciences, 4301 W. Markham, Slot 505, Little Rock, AR, 72205, USA.
authors.forename: Laura
authors.initials: L
authors.lastname: MacDonald
authors.forename: Giulia
authors.initials: G
authors.lastname: Baldini
authors.forename: Brian
authors.initials: B
authors.lastname: Storrie
mesh_terms: D018613:Microscopy, Confocal / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D021381:Protein Transport; D011506:Proteins / Q000378:metabolism*
chemical_list: D011506:Proteins