Toolkit

Our research integrates molecular virology, cell biology, single-molecule biophysics, and computational modeling. As a result, trainees in the Ivanovic lab learn not only how to think deeply as virologists, but also acquire an unusual combination of interdisciplinary skills that together permit them to seek answers to sophisticated questions. Opportunities for learning include how to manipulate viral genomes to generate ‘designer’ viruses, how to use viruses to learn new cell biology, how to use, design and build specialized microscopes, and how to write computer codes simulating single-molecule events in virus system models.

Building designer viruses using reverse genetics systems and other tricks
Generating viruses from plasmid DNA
Generating viruses using recombinant proteins
Visualizing virus assembly in real time
Transmission electron microscopy to view purified virus particles or viral proteins.
Pleomorphic influenza virus particles
Spherical influenza virus particles
Coronavirus NL-63
Single RNA molecule fluorescence in situ hybridization (RNAscope) to quantify virus attachment to cells, mRNA expression, and genome replication.
Viral genomic RNA is detected with a sense-RNA probe. Each red dot corresponds to a single viral genome
Overlay of phase contrast (cells) and fluorescence (includes Dapi for nuclei) images

(Left) Influenza virus binding to cells at the onset of infection. (Bottom) Viral mRNA expression after 12 hours of infection.

mRNA (green) and Dapi (blue)
Home-built Total Internal Reflection Fluorescence (TIRF) microscope for single-virus particle and single-molecule imaging. As our research evolves, we update the design of our microscopes to meet the changing experimental needs.
Viral membrane fusion visualized at the level of single virions
Spherical flu, no inhibitor
Pleomorphic flu, no inhibitor
Spherical flu, fusion inhibitor
Pleomorphic flu, fusion inhibitor
Stochastic simulations of the molecular events at the interface between the viral and target cell membranes.
Model of molecular events leading to membrane fusion
Stochastic simulation model of membrane fusion
Simulation results: filamentous particles resist HA inactivation