Work

Sample Preparation for Single Virion Atomic Force Microscopy and Super-resolution Fluorescence Imaging

Year: 2014

Type: article

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.

Source: Journal of Visualized Experiments

DOI: 10.3791/51366-v

Authors: Jeffery A Hodges, Saveez Saffarian

Institution: University of Utah

Language: English

Topic: Force Microscopy Techniques and Applications

Subfield: Atomic and Molecular Physics, and Optics

Field: Physics and Astronomy

Domain: Physical Sciences

Open Access status: bronze

Funder: National Science Foundation

Award: Developing Methods for High-resolution Measurements of Enveloped Virus Budding in Live Cells