The Vutara single molecule localization system has been a critical instrument for understanding viral particles. Viral particles are typically much smaller than the diffraction limit of light (<200 nm), making single molecule localization microscopy the best suited fluorescence technique for resolving virus particle structural details or determining localization of virus components with the cellular machinery. Below we highlight the key features of the Vutara for virus research.
- Super resolved images using proprietary biplane single molecule localization, achieving at least 20 nm XY and 50 nm Z precision.
- The only 3D single molecule system capable of imaging multiple sample types, from purified virions to tissue sections and whole model organisms.
- High speed acquisition: ideal for live imaging, particle tracking and rapid data acquisition.
- Integrated fluidics: multiplexed imaging of the proteome, genome or live cell applications.
- Powerful acquisition software with real time single molecule localization.
- Powerful visualization and analysis software package provides a complete statistics tool set.
Below we highlight virus research performed on the Vutara. The unique ability to perform single molecule localization of virus samples at both the coverslip and deep within tissue sections makes the Vutara the only system capable of imaging virus particle structures, virus particle host cell interactions, and effects of virus infection on cell biology on the same microscope. At the bottom of the page you can find some highlighted virus research papers performed with the Vutara super resolution microscope.
Alonas, E., Lifland, A.W., Gudheti, M., Vanover, D., Jung, J., Zurla, C., Kirschman, J., Fiore, V.F., Douglas, A., Barker, T.H., Yi, H., Wright, E.R., Crowe, J.E., Santangelo, P.J., 2014. Combining Single RNA Sensitive Probes with Subdiffraction-Limited and Live-Cell Imaging Enables the Characterization of Virus Dynamics in Cells. ACS Nano 8, 302–315. doi.org/10.1021/nn405998v
The authors developed tools to study the early infectivity and replication of enveloped viruses.
- The authors developed MTRIPs (multiply labeled tetravalent RNA imaging probes). A method to live label hRSV viral genomes.
- The MTRIP technique enabled the simultaneous super resolution imaging of proteins and the viral genome; not possible with conventional fluorescence in situ hybridization techniques (FISH).
- The authors used the Vutara 352 to determine the distribution of viral proteins along the viral gRNA. Only single molecule localization microscopy gives the resolution to image these sub-300 nm particles
Milrot, E., Shimoni, E., Dadosh, T., Rechav, K., Unger, T., Etten, J.L.V., Minsky, A., 2017. Structural studies demonstrating a bacteriophage-like replication cycle of the eukaryote-infecting Paramecium bursaria chlorella virus-1. PLOS Pathogens 13, e1006562. doi.org/10.1371/journal.ppat.1006562
The authors used the Vutara to determine the effects of viral infection on cytoskeletal structure. From this they determined that the actin cytoskeleton plays a critical role in viral infectivity.
- The authors used super-resolution imaging to monitor how the microtubule and actin cytoskeleton changed over the course of viral infection.
- During infection, the microtubule network became more fragmented and disappeared from the center of the cell.
- During infection, the actin cytoskeleton loses its fine structure at the periphery of the cell and forms a shell around the rounded edge of the cell.
- Pharmacological experiments and other experiments revealed that disruption of the microtubule network had little effect on virion production, while disruption of the actin cytoskeleton reduced virion production.