Methods · 2016 · 12 citations · 12 references
Life SciencesBiologyCell BiologyCell AdhesionCytoskeletonStructural BiologyCell-matrix AdhesionCellular UltrastructureCytoskeletal ArchitectureExtracellular MatrixMicroscopic Cell MorphologyProtein LocalizationCytoskeletal ImagingCytoskeletal VisualizationImage-based Cell BiophysicsMembrane Protein LocalizationMicroscopyRing StructureSubcellular Localization Mapping
Podosomes are adhesive structures formed on the plasma membrane abutting the extracellular matrix of macrophages, osteoclasts, and dendritic cells. They consist of an f-actin core and a ring structure composed of integrins and integrin-associated proteins. The podosome ring plays a major role in adhesion to the underlying extracellular matrix, but its detailed structure is poorly understood. Recently, it has become possible to study the nano-scale structure of podosome rings using localization microscopy. Unlike traditional microscopy images, localization microscopy images are reconstructed using discrete points, meaning that standard image analysis methods cannot be applied. Here, we present a pipeline for podosome identification, protein position calculation, and creating a podosome ring model for use with localization microscopy data.
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Martin A. Fischler, Robert C. Bolles · Communications of the ACM · 1981 · 25.5K citations · Full text
Imaging Intracellular Fluorescent Proteins at Nanometer Resolution
Eric Betzig, George H. Patterson, Rachid Sougrat et al. · Science · 2006 · 8.9K citations