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Sample Preparation for Single Virion Atomic Force Microscopy and Super-resolution Fluorescence Imaging
Published on: January 2, 2014
Fluorescence molecular painting of enveloped viruses
Christoph Metzner1, Feliks Kochan, John A Dangerfield
1Institute of Virology, University of Veterinary Medicine, Vienna, Veterinarplatz 1, 1210 Vienna, Austria. christoph.metzner@vetmeduni.ac.at
Abstract:
In this study, we describe a versatile, flexible, and quick method to label different families of enveloped viruses with glycosylphosphatidylinositol-modified green fluorescent protein, termed fluorescence molecular painting (FMP). As an example for a potential application, we investigated virus attachment by means of flow cytometry to determine if viral binding behavior may be analyzed after FMP of enveloped viruses. Virus attachment was inhibited by using either dextran sulfate or by blocking attachment sites with virus pre-treatment. Results from the FMP-flow cytometry approach were verified by immunoblotting and enzyme-linked immunosorbent assay. Since the modification strategy is applicable to a broad range of proteins and viruses, variations of this method may be useful in a range of research and applied applications from bio-distribution studies to vaccine development and targeted infection for gene delivery.

