title: Investigation of podosome ring protein arrangement using localization microscopy images.
abstract: 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.
authors.affiliation: Randall Division of Cell and Molecular Biophysics, King's College London, London, UK.
authors.forename: Adela D
authors.initials: AD
authors.lastname: Staszowska
authors.affiliation: Randall Division of Cell and Molecular Biophysics, King's College London, London, UK.
authors.forename: Patrick
authors.initials: P
authors.lastname: Fox-Roberts
authors.affiliation: Randall Division of Cell and Molecular Biophysics, King's College London, London, UK.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Foxall
authors.affiliation: Randall Division of Cell and Molecular Biophysics, King's College London, London, UK.
authors.forename: Gareth E
authors.initials: GE
authors.lastname: Jones
authors.affiliation: Randall Division of Cell and Molecular Biophysics, King's College London, London, UK. Electronic address: susan.cox@kcl.ac.uk.
authors.forename: Susan
authors.initials: S
authors.lastname: Cox
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism / Q000648:ultrastructure*; D002232:Carbocyanines / Q000737:chemistry; D002465:Cell Movement; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000378:metabolism / Q000648:ultrastructure; D005109:Extracellular Matrix / Q000378:metabolism / Q000648:ultrastructure*; D005347:Fibroblasts / Q000378:metabolism / Q000648:ultrastructure; D005456:Fluorescent Dyes / Q000737:chemistry; D015870:Gene Expression; D017930:Genes, Reporter; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism / Q000648:ultrastructure*; D008856:Microscopy, Fluorescence / Q000379:methods*; D010010:Osteoclasts / Q000378:metabolism / Q000648:ultrastructure; D051419:Paxillin / Q000235:genetics / Q000378:metabolism; D000069261:Podosomes / Q000378:metabolism / Q000648:ultrastructure*; D013194:Staining and Labeling / Q000379:methods; D016608:Talin / Q000235:genetics / Q000378:metabolism; D016596:Vinculin / Q000235:genetics / Q000378:metabolism; D000097573:Red Fluorescent Protein
chemical_list: D002232:Carbocyanines; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D051419:Paxillin; D016608:Talin; D016596:Vinculin; D000097573:Red Fluorescent Protein; C569686:Alexa Fluor 647; C496061:PXN protein, human; C488955:VCL protein, human; C089187:cyanine dye 3
keywords: Image analysis; Localization microscopy; Pattern recognition; Podosomes; Super-resolution microscopy