title: Photoactivated Localization Microscopy (PALM) of adhesion complexes.
abstract: Key to understanding a protein's biological function is the accurate determination of its spatial distribution inside a cell. Although fluorescent protein markers allow the targeting of specific proteins with molecular precision, much of this information is lost when the resultant fusion proteins are imaged with conventional, diffraction-limited optics. In response, several imaging modalities that are capable of resolution below the diffraction limit (∼200 nm) have emerged. Here, both single- and dual-color superresolution imaging of biological structures using photoactivated localization microscopy (PALM) are described. The examples discussed focus on adhesion complexes: dense, protein-filled assemblies that form at the interface between cells and their substrata. A particular emphasis is placed on the instrumentation and photoactivatable fluorescent protein (PA-FP) tags necessary to achieve PALM images at ∼20 nm resolution in 5 to 30 min in fixed cells.
authors: National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland.
authors: Hari
authors: H
authors: Shroff
authors: Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, Virginia.
authors: Helen
authors: H
authors: White
authors: Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, Virginia.
authors: Eric
authors: E
authors: Betzig
mesh_terms: D008856:Microscopy, Fluorescence / Q000379:methods*; D046912:Multiprotein Complexes / Q000378:metabolism*; D012984:Software
chemical_list: D046912:Multiprotein Complexes