High-Speed 3D Tracking and Imaging Microscopy Captures Early Events of the Virus-Cell Interaction
Courtney "CJ" Johnson (Janelia)
Abstract: Single-particle tracking methods have shifted the landscape of physical virology from bulk methods requiring ensemble averaging to the single virus level, yielding new quantitative information about the virus infection process. However, existing methods are unable to study the critical extracellular phase of this process. In this phase, where the virus diffuses to meet the cell surface, the disparate spatial scales between viruses and cells, combined with the rapid motion of viruses makes studying their interaction at high speed an impossible task for any single microscope. Here we present 3D Tracking and Imaging microscopy (3D-TrIm): a multi-modal instrument that simultaneously acquires co-registered volumes featuring the trajectories of fluorescent virus-like particles and multi-photon volumetric images of the surrounding live-cell environment. This approach yields an unprecedented 3D view into the interaction of virions at the cellular interface. We demonstrate the power of our super-temporally-resolved tracking to reveal previously invisible interactions and trace out nanoscale features on the cell membrane. Finally, we extend these studies into multi-layer epithelial tissue.
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