title: Sample preparation for single virion atomic force microscopy and super-resolution fluorescence imaging.
abstract: Immobilization of virions to glass surfaces is a critical step in single virion imaging. Here we present a technique adopted from single molecule imaging assays which allows adhesion of single virions to glass surfaces with specificity. This preparation is based on grafting the surface of the glass with a mixture of PLL-g-PEG and PLL-g-PEG-Biotin, adding a layer of avidin, and finally creating virion anchors through attachment of biotinylated virus specific antibodies. We have applied this technique across a range of experiments including atomic force microscopy (AFM) and super-resolution fluorescence imaging. This sample preparation method results in a control adhesion of the virions to the surface.
authors.affiliation: Department of Physics & Astronomy, University of Utah.
authors.forename: Jeffery A
authors.initials: JA
authors.lastname: Hodges
authors.forename: Saveez
authors.initials: S
authors.lastname: Saffarian
mesh_terms: D001360:Avidin / Q000737:chemistry; D001710:Biotin / Q000737:chemistry; D002232:Carbocyanines / Q000737:chemistry; D018625:Microscopy, Atomic Force / Q000379:methods*; D061848:Optical Imaging / Q000379:methods*; D011092:Polyethylene Glycols / Q000737:chemistry; D011107:Polylysine / Q000031:analogs & derivatives / Q000737:chemistry; D014771:Virion / Q000737:chemistry*; D014773:Virology / Q000379:methods*
chemical_list: C569686:Alexa Fluor 647; D002232:Carbocyanines; C443714:polylysine-graft-(poly(ethylene glycol)); D001360:Avidin; D011107:Polylysine; D011092:Polyethylene Glycols; D001710:Biotin