title: Localization and quantification of epidermal growth factor receptors on single cells by confocal laser scanning microscopy.
abstract: We have established a method for quantifying binding of fluorescence-labeled growth factors to their receptors on single cells in situ with the confocal laser scanning microscope (CLSM). Biotinylated epidermal growth factor (EGF) coupled to phycoerythrin-labeled anti-biotin was used to compare the levels of fluorescence on three different cell types for which the number of EGF factors was known from Scatchard analysis of [125I]-EGF binding. The results showed that as few as 10,000 receptors/cell were detectable above back-ground. This method will provide a rapid and quantifiable alternative to autoradiography for ligand binding to single cells in situ.
authors.affiliation: Hubrecht Laboratory, Netherlands Institute for Developmental Biology, University of Utrecht.
authors.forename: M J
authors.initials: MJ
authors.lastname: Good
authors.forename: W J
authors.initials: WJ
authors.lastname: Hage
authors.forename: C L
authors.initials: CL
authors.lastname: Mummery
authors.forename: S W
authors.initials: SW
authors.lastname: De Laat
authors.forename: J
authors.initials: J
authors.lastname: Boonstra
mesh_terms: D001710:Biotin / Q000378:metabolism; D002478:Cells, Cultured; D004815:Epidermal Growth Factor / Q000378:metabolism; D066246:ErbB Receptors / Q000378:metabolism* / Q000648:ultrastructure; D005453:Fluorescence; D007834:Lasers; D008854:Microscopy, Electron / Q000379:methods
chemical_list: D004815:Epidermal Growth Factor; D001710:Biotin; D066246:ErbB Receptors