title: Epidermal growth factor receptor subunit locations determined in hydrated cells with environmental scanning electron microscopy.
abstract: Imaging single epidermal growth factor receptors (EGFR) in intact cells is presently limited by the available microscopy methods. Environmental scanning electron microscopy (ESEM) of whole cells in hydrated state in combination with specific labeling with gold nanoparticles was used to localize activated EGFRs in the plasma membranes of COS7 and A549 cells. The use of a scanning transmission electron microscopy (STEM) detector yielded a spatial resolution of 3 nm, sufficient to identify the locations of individual EGFR dimer subunits. The sizes and distribution of dimers and higher order clusters of EGFRs were determined. The distance between labels bound to dimers amounted to 19 nm, consistent with a molecular model. A fraction of the EGFRs was found in higher order clusters with sizes ranging from 32-56 nm. ESEM can be used for quantitative whole cell screening studies of membrane receptors, and for the study of nanoparticle-cell interactions in general.
authors.affiliation: INM - Leibniz Institute for New Materials, 66123 Saarbrücken, Germany.
authors.forename: Diana B
authors.initials: DB
authors.lastname: Peckys
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Baudoin
authors.forename: Magdalena
authors.initials: M
authors.lastname: Eder
authors.forename: Ulf
authors.initials: U
authors.lastname: Werner
authors.forename: Niels
authors.initials: N
authors.lastname: de Jonge
mesh_terms: D000818:Animals; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D004815:Epidermal Growth Factor / Q000737:chemistry / Q000378:metabolism; D066246:ErbB Receptors / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D053768:Metal Nanoparticles / Q000737:chemistry; D008855:Microscopy, Electron, Scanning; D008958:Models, Molecular; D011487:Protein Conformation; D055503:Protein Multimerization; D021122:Protein Subunits; D021381:Protein Transport
chemical_list: D021122:Protein Subunits; D004815:Epidermal Growth Factor; D066246:ErbB Receptors
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