Cost-effective high-speed, three-dimensional live-cell imaging of HIV-1 transfer at the T cell virological synapse

Abstract

The availability of cost-effective, highly portable, and easy to use high-resolution live-cell imaging systems could present a significant technological break-through in challenging environments, such as high-level biosafety laboratories or sites where new viral outbreaks are suspected. We describe and demonstrate a cost-effective high-speed fluorescence microscope enabling the live tracking of virus particles across virological synapses that form between infected and uninfected Tcells. The dynamics of HIV-1 proteins studied at the cellular level and the formation of virological synapses in living Tcells reveals mechanisms by which cell-cell interactions facilitate infection between immune cells. Dual-color 3D fluorescence deconvolution microscopy of HIV-1 particles at frames rates of 100 frames per second allows us to follow the transfer of HIV-1 particles across the Tcell virological synapse between living Tcells. We also confirm the successful transfer of virus by imaging Tcell samples fixed at specific time points during cell-cell virus transfer by super-resolution structured illumination microscopy.