Imaging Core

Directors: Zandrea Ambrose and Simon Watkins

The Imaging Core visualizes the intracellular dynamics of HIV-1 components during the processes of HIV-1 CA core uncoating, RTxn, and PIC nuclear entry, by live-cell and high-resolution fixed cell microscopy, using existing and novel technologies. The Core will use labeled virions and host cell proteins of PCHPI-selected interest for conducting imaging studies and will develop novel techniques to track post-entry events, visualizing viral protein and nucleic acids that interact with host proteins using a complete array of super-resolution, optical sectioning, and wide field approaches in close collaboration with the technology development program (Peijun Zhang).

The Core is primarily housed within the Center for Biologic Imaging (CBI), which is equipped to perform a continuum of optical methods, including all types of microscopy essential to the PCHPI. The Imaging Core will produce and biologically characterize fluorescently labeled viruses; develop vectors containing tagged cellular proteins for production of transiently transfected and stable cell lines; isolate and image primary human T cells, macrophages, and dendritic cells; conduct live-cell single particle and single molecule imaging in BSL2/2+ facilities; and apply super-resolution single particle/molecule fluorescence imaging to visualize viral and host cell proteins.

The Imaging Core will work closely with the cryoEM Core to carry out studies in the Correlative Single Particle Imaging Program of the PCHPI. See: 

Direct visualization of HIV-1 with correlative live-cell microscopy and cryo-electron tomography. Jun S, Ke D, Debiec K, Zhao G, Meng X, Ambrose Z, Gibson GA, Watkins SC, and Zhang P. (2011) Structure 19 (11): 1573-1581. PubMed

Correlative microscopy for 3D structural analysis of dynamic interactions. Jun S, Zhao G, Ning J, Gibson GA, Watkins SC, Zhang P. J Vis Exp. 2013 Jun 24;(76). PubMed

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